JP2005518185A - Novel protein and nucleic acid encoding it - Google Patents

Abstract

The present invention provides novel isolated polypeptides and small molecule target polypeptides encoded by the polynucleotides. Disclosed are antibodies, polynucleotides or antibodies that immunospecifically bind to a novel small molecule target polypeptide or derivative, polypeptide variant, mutant or fragment. Disclosed are methods that utilize small molecule target polypeptides, polynucleotides, and antibodies in a wide range of disease states. More specifically, the present invention relates to recombinantly expressed and / or endogenously expressed proteins in various screening methods aimed at identifying therapeutic antibodies and therapeutic small molecules associated with disease. Provide a method to use.

Description

Detailed Description of the Invention

(Technical field)
The present invention relates to novel polypeptides that are targets for small molecule drugs and have characteristic properties related to biochemical stimuli or physiological responses in cells, tissues, organs or organisms. More particularly, the novel polypeptide is the gene product of a novel gene or a specific biologically active fragment or derivative thereof. Methods of use include diagnostic or prognostic assays, as well as methods for treating various pathological conditions.

(Background technology)
Eukaryotic cells feature biochemical and physiological methods that are delicately balanced to achieve cell maintenance and proliferation under normal conditions. When such cells are components of multicellular organisms such as vertebrates, or more particularly organisms such as mammals, the regulation of biochemical and physiological methods involves sophisticated signaling pathways. . Often, such signaling pathways involve extracellular signaling proteins, cellular receptors that bind to signaling proteins, and signaling components that are localized within the cell.

  Signal transduction proteins can be classified as endocrine effectors, paracrine effectors or autocrine effectors. Endocrine effectors are signaling molecules that are secreted into the circulatory system by a particular organ, which is then transported to a remote target organ or tissue. Target cells contain receptors for endocrine effectors, and when the endocrine effectors bind, a signaling cascade is induced. Paracrine effectors involve secretory cells and recipient cells in close proximity to each other, for example, two different classes of cells within the same tissue or organ. The first class of cells secretes the paracrine effector, which then reaches the second class of cells, for example by diffusion through the extracellular fluid. The second class of cells contains receptors for paracrine effectors, and effector binding induces a signaling cascade that results in a corresponding biochemical or physiological effect. Autocrine effectors have a high degree of similarity to paracrine effectors, except that the same type of cells that secrete autocrine effectors also contain receptors. Thus, an autocrine effector binds to a receptor on the same cell or on the same adjacent cell. The binding process then causes a characteristic biochemical or physiological effect.

  Signal transduction processes for cells and tissues include, but are not limited to, induction of cell or tissue proliferation, inhibition of growth or proliferation, induction of cell or tissue differentiation or maturation, and cell or tissue differentiation or maturation. Can cause a variety of effects including inhibition.

  Many pathologies involve dysregulation of the expression of important effector proteins. In certain classes of conditions, dysregulation manifests as a reduced or suppressed level of protein effector synthesis and secretion. In a clinical setting, a subject may be suspected of suffering from an abnormality caused by a decrease or suppression of a protein effector level of interest. Therefore, there is a need to assay the level of protein effector of interest in a biological sample from such a subject and compare that level to that characteristic of a non-pathological condition. There is a further need for methods of treatment of pathologies caused by increased or excessive levels of protein effectors of interest based on administering antibodies to a subject. Administration of the effector or protein effector to a subject in need thereof that is useful in the treatment of a disease state is preferably performed by administration of another small molecule drug product. Therefore, there is a need for methods of treating pathologies caused by reduced or suppressed levels of the protein effectors.

  Small molecule targets have been implicated in disease states and pathologies. These targets can be proteins, in particular enzyme proteins, which act with small molecule drugs for the purpose of changing the target function and achieving the desired result. Cell studies, animal studies and clinical studies can be performed to elucidate genes that contribute to the pathogenesis and pathogenicity of the condition. Here, the small molecule target may be involved in various physiological, pharmacological, or original conditions. These studies utilize the core technology at CuraGen Corporation, and are not limited to different gene expression, protein-protein interactions, large-scale sequences of expressed genes and the relevance of such gene variants, without limitation Find polymorphisms (SNPs) or splice variants within or between biological samples derived between samples from experimental and coping groups. The goal of such research is to identify potential means of therapeutic treatment to prevent outcome or cure the condition.

  Novel therapeutic agents for treating diseases, lesions and other above conditions or abnormal conditions other than normal or normal conditions diagnosed as, or at risk for, mammalian tissue It is important to identify. Such methods include at least identifying target components in the injured tissue or organ and identifying candidate therapeutic agents that modulate the functional properties of the target. The target component can be a biological macromolecule involved in a disease or pathology. Common targets can be lipids such as nucleic acids, polysaccharides, complex lipids or glycolipids; in addition, the target can be a semi-intracellular or extracellular structure consisting of more than one class of macromolecules . Once such targets were identified. It was used in screening assays to identify preferred candidate therapeutics from a large population of substances or components.

  In many cases, the purpose of such screening assays is to identify small molecule candidates; this is generally approached by a combination of methodologies for generating a population of substances to be tested. Implementation of high-throughput screening methods is advantageous when working with large libraries of compound combinations.

(Summary of the Invention)
The present invention is based in part on the discovery of nucleic acid sequences that encode novel polypeptides. The novel nucleic acids and polypeptides, as well as their nucleic acids and polypeptides, and derivatives, homologs, analogs and fragments thereof are hereinafter collectively referred to as “NOVX” nucleic acid or polypeptide sequences, and SEQ ID NOs: 2n-1 (n is 1 to 1). Represents a nucleotide sequence selected from the group consisting of SEQ ID NO: 538, or a polypeptide representing the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539.

  In one aspect, the invention provides an isolated peptide comprising a mature form of NOVX amino acid. One example is a mature variant of the NOVX amino acid. Here, any mature amino acid has been changed to a different amino acid, provided that less than 15% of the amino acid residues in the sequence are changed. An amino acid is, for example, a NOVX amino acid sequence, or a NOVX amino acid that has been changed to a different amino acid, provided that any amino acid specified in the selected sequence is changed in less than 15% of the amino acid residues in the sequence It may be a variant of the sequence. The present invention also includes any of these fragments. In another aspect, the invention also includes an isolated nucleic acid encoding a NOVX polypeptide, or a fragment, homolog, analog or derivative thereof.

  The invention also includes NOVX polypeptides that are naturally occurring allelic variants of the NOVX sequence. In one embodiment, an allelic variant comprises an amino acid sequence that is a translation of a nucleic acid sequence that differs from the NOVX nucleic acid sequence by one nucleotide. In another embodiment, the NOVX polypeptide is a variant polypeptide described therein, in which any amino acid identified in the selected sequence has been altered to produce a conservative substitution. In one embodiment, the present invention discloses a method for measuring the presence or amount of NOVX polypeptide in a sample. The method comprises preparing a sample; introducing into the sample an antibody that immunospecifically binds to the polypeptide; and measuring the presence or amount of an antibody that binds to the NOVX polypeptide, thereby causing NOVX polyion in the sample. Measuring the presence or amount of the peptide. In another embodiment, the present invention provides a method of determining the presence or predisposition of a disease associated with an altered level of NOVX polypeptide in a mammalian subject. The method measures the expression level of the polypeptide in the sample from the first mammalian subject; and the amount of the polypeptide in the first stage sample has no disease or predisposition Comparing the amount of polypeptide present in a control sample from a second mammalian subject, wherein the level of expression of the polypeptide in the first subject when compared to the control sample. Changes include methods that indicate the presence or predisposition of the disease.

  In a further embodiment, the invention includes a method of identifying a reagent that binds to a NOVX polypeptide. The method includes introducing a polypeptide into the reagent; and determining whether the reagent binds to the polypeptide. In various embodiments, the reagent is an intracellular receptor or a downstream effector.

  In another embodiment, the present invention provides a method of identifying potential therapeutic agents for use in the treatment of lesions. Here, the lesion is associated with abnormal expression of NOVX polypeptide or abnormal physiological interaction. The method expresses a NOVX polypeptide and provides a cell having a property or function attributable to the polypeptide; contacting the cell with a composition comprising a candidate substance; and a property or function resulting from the polypeptide The substance is potential if the change observed in the presence of the substance is not observed when the cell is contacted with the composition in the absence of the substance. A step identified as a therapeutic agent. In another embodiment, the present invention describes a method of screening for modulators of the activity or potential activity or predisposition of a lesion associated with a NOVX polypeptide. The method includes the following steps. Administering a test compound to a test animal at an increased risk of a lesion associated with the NOVX polypeptide, wherein the test animal recombinantly expresses the NOVX polypeptide. This method measures the activity of a NOVX polypeptide in a test animal after the compound administration phase; and the activity of the protein in the test animal determines the activity of the NOVX polypeptide in a control animal that has not been administered the polypeptide. Where a change in the activity of the NOVX polypeptide in the test animal as compared to the control animal indicates that the compound is a modulator of the potential activity or predisposition to lesions associated with the NOVX polypeptide. Including that. In one embodiment, the test animal is a recombinant test animal that expresses the protein transgene or expresses the transgene under the control of an increased level of promoter compared to the wild type. Here, the promoter is not the natural gene promoter of the transgene. In another aspect, the invention includes a method of modulating the activity of a NOVX polypeptide. The method includes introducing a cell sample expressing a NOVX polypeptide with a compound that binds to the polypeptide in an amount sufficient to modulate the activity of the polypeptide.

  The present invention also includes isolated nucleic acids, fragments, homologues, analogs or derivatives thereof that encode NOVX polypeptides. In a preferred embodiment, the nucleic acid molecule comprises the nucleotide sequence of a naturally occurring allelic variant. In another embodiment, the nucleic acid encodes a variant polypeptide having the polypeptide sequence of a naturally occurring polypeptide variant. In another embodiment, the nucleic acid molecule differs from the NOVX nucleic acid sequence by a single nucleotide. In certain embodiments, the NOVX nucleic acid molecule hybridizes under stringent conditions to a nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538. In another aspect, the present invention provides a cell that expresses a vector or NOVX nucleotide sequence.

  In one embodiment, the present invention discloses a method for measuring the activity of a NOVX polypeptide. The method includes introducing a cell sample expressing a NOVX polypeptide with a compound that binds to the polypeptide in an amount sufficient to modulate the activity of the polypeptide. In another embodiment, the invention includes an isolated NOVX nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising a NOVX amino acid sequence or a mature variant of the NOVX amino acid sequence. Here, any amino acid in the mature form of the selected sequence has been converted to a different amino acid, provided that less than 15% of the amino acid residues in the mature sequence are changed. In another embodiment, the invention includes an amino acid sequence that is a variant of the NOVX amino acid sequence. Here, any particular amino acid in the selected sequence has been changed to a different amino acid, provided that less than 15% of the amino acid residues in the sequence are changed.

  In one aspect, the invention discloses a NOVX nucleic acid fragment that encodes at least a portion of a NOVX polypeptide or any variant of the polypeptide. Here, any amino acid in the selected sequence is changed to a different amino acid, provided that less than 10% of the amino acid residues in the sequence are changed. In another embodiment, the present invention discloses NOVX nucleic acid molecules that encode variant polypeptides having the polypeptide sequence of a naturally occurring polypeptide variant. In another embodiment, the present invention discloses NOVX nucleic acids that differ by a single nucleotide from a NOVX nucleic acid sequence.

  In another embodiment, the invention includes a NOVX nucleic acid in which one or more nucleotides in the NOVX nucleotide sequence are changed to different nucleotides provided that less than 15% of the nucleotides are changed. In one aspect, the present invention discloses nucleic acid fragments and nucleic acid fragments of NOVX nucleotide sequences. Here, one or more nucleotides in the NOVX nucleotide sequence have been changed from one selected from the group consisting of the selected sequence to a different nucleotide, provided that less than 15% of the nucleotides are changed. In another embodiment, the present invention comprises a nucleic acid molecule. Here, the nucleic acid molecule hybridizes under stringent conditions to the NOVX nucleotide sequence or a complementary sequence of the NOVX nucleotide sequence. In one embodiment, the present invention comprises a nucleic acid molecule. Here, the sequence has been altered so that less than 15% of the nucleotides in the coding sequence differ from the NOVX nucleotide sequence or fragments thereof.

  In a further aspect, the present invention includes a method for determining the presence or amount of NOVX nucleic acid in a sample. The method comprises preparing a sample; introducing the sample into a probe that binds to a nucleic acid molecule; and measuring the presence or amount of a probe that binds to a NOVX nucleic acid molecule, thereby the presence or absence of a NOVX nucleic acid molecule in the sample. Comprising a step consisting of measuring a quantity. In one embodiment, the presence or amount of a nucleic acid molecule is used as a marker for cell type or tissue type.

  In another aspect, the present invention discloses a method of determining the presence or predisposition of a disease associated with an altered level of NOVX nucleic acid molecule in a first mammalian subject. The method measures the amount of NOVX nucleic acid in the sample from the first mammalian subject; and the amount of nucleic acid in the sample of step (a) may be disease free or predisposed Comparing to the amount of NOVX nucleic acid present in a control sample from a known second mammalian subject; wherein the change in the level of nucleic acid in the first subject as compared to the control sample is a disease Showing the presence or predisposition of.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are hereby incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative and not intended to be limiting.
Other features and advantages of the invention will be apparent from the following invention disclosure and claims.

(Disclosure of the Invention)
The present invention provides novel nucleotides and polypeptides encoded thereby. The present invention includes novel nucleic acid sequences, their encoded polypeptides, antibodies, and other related compounds. As used herein, sequences are collectively referred to as “NOVX nucleic acids” or “NOVX polynucleotides” and the corresponding encoded polypeptides are referred to as “NOVX polypeptides” or “NOVX proteins”. Unless otherwise stated, “NOVX” is meant to refer to any novel sequence disclosed herein. Table 1 summarizes the NOVX nucleic acids and their encoded polypeptides.

Table 1: Sequences and corresponding sequence numbers

Continuation of Table 1

Continuation of Table 1

Continuation of Table 1

Continuation of Table 1

Continuation of Table 1

  Table 1 shows the homology of NOVX polypeptides with known protein families. Thus, the nucleic acid polynucleotides, antibodies and related compounds according to the present invention corresponding to NOVX identified in the first column of Table 1 are, for example, lesions associated with known protein families identified in the fifth column of Table 1. And useful in therapeutic and diagnostic applications related to disease.

  NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and situations. Various NOVX nucleic acids and polypeptides according to the present invention are useful as novel members of a protein family according to the existence of domains and sequence relationships with said proteins. In addition, NOVX nucleic acids and polypeptides can be used to identify proteins that are members of the family to which the NOVX polypeptide belongs.

  Consistent with other known members of the protein family identified in column 5 of Table 1, the NOVX polypeptides of the present invention exhibit homology to and characterize other members of such protein families. Contains a domain that Details of sequence relatedness analysis and domain analysis for each NOVX are shown in Example A.

  NOVX nucleic acids and polypeptides can be used to screen for molecules that inhibit or promote NOVX activity or function. In particular, the nucleic acids and polypeptides according to the invention can be used as targets for the identification of small molecules that modulate or inhibit diseases associated with the protein families listed in Table 1.

NOVX nucleic acids and polypeptides are also useful for detecting specific cell types. Details of expression analysis for each NOVX are shown in Example C. Accordingly, NOVX nucleic acids, polypeptides, antibodies and related compounds according to the present invention have diagnostic and therapeutic applications in the detection of various diseases having different expression in normal tissues versus diseased tissues, eg, various cancers. .
Additional uses for NOVX nucleic acids and polypeptides according to the present invention are disclosed herein.

  The present invention is based on the identification of biological macromolecules that are differentially regulated in lesions, diseases, or abnormal conditions or conditions. Such lesions or diseases include endocrine diseases, cancer, various tumors and tumorigenesis, inflammatory diseases, central nervous system diseases, and metabolic diseases including diseases associated with similar abnormal conditions or conditions. In a very significant embodiment of the invention, the biological macromolecules involved in the pathology and condition are proteins and polypeptides, and in such cases of the invention also relate to the nucleic acids that encode them. Methods that can be used to identify related biological macromolecules include any method that detects differential expression of nucleic acids encoding proteins and polypeptides associated with disease, as well as the respective proteins and their own polys. A method of detecting a peptide is included. Significant methods that have been used in accordance with the present invention include CuraGen ™ technology and SeqCalling ™ technology, US serial number 5,871,697 and US serial number filed October 13, 1999, respectively. No. 09 / 417,386, each of which is incorporated herein by reference in its entirety. GeneCalling® is also described in Shimkets, et al., “Gene expression analysis by transcript profiling coupled to a gene database query” Nature Biotechnology 17: 198-803 (1999).

  The present invention provides polypeptides and nucleotides that have been identified as having a novel association with a disease or pathology in a mammal, or having an abnormal state or condition. The present invention relates to proteins comprising naturally occurring polypeptides, proprotein precursor forms, or mature forms of polypeptides or proteins involved as targets for therapeutic reagents in the treatment of various diseases, pathologies, abnormal conditions and conditions And a set of polypeptides are further identified. A target that can be used in any of a variety of screening methods to identify candidate therapeutic agents that interact with the target exerts the desired or favorable effect. Candidate therapeutic agents are identified in an important embodiment of the invention by screening a large collection of substances or compounds. Such a collection may include a combinatorial library of substances and compounds, at least individual members of a subset of substances or compounds being each involved by a variation of a simple structure based on a specific limit structure or basic chemical structure. Variations are not limited to the examples, but variations in length and identity of the basic framework of the linking atom; composition and properties of number changes, cyclic structures, bridge structures, alicyclic and aromatic rings; And may include intermediate atoms and a basic framework of linking atoms, or a substituted atom or group bonded at a particular position to a ring structure, bridge structure, alicyclic ring, or aromatic ring.

  The polypeptides and proteins described herein are used as targets in screening methods and include naturally occurring polypeptides or precursor products, or proproteins. Naturally occurring polypeptides, precursors, or proproteins include, for example, full-length gene products encoded by the corresponding genes. Naturally occurring polypeptides also include polypeptides, precursors, or proproteins encoded by the open reading frames described herein. A “mature” form of a polypeptide or protein occurs as a result of one or more naturally occurring processing steps that occur within a cell, including a host cell. Processing steps that occur as gene products occur, for example, via cleavage of the amino terminal methionine residue encoded by the start codon of the open reading frame, or proteolytic cleavage of the signal peptide or leader sequence. Thus, a mature form resulting from a precursor polypeptide or protein having residues 1-N where residue 1 is the N-terminal methionine has residues 2 through N residues. Alternatively, a mature form resulting from a precursor polypeptide or protein having residues 1-N cleaved from the amino-terminal signal sequence from residues 1-M includes residues remaining from residue M + 1 to residue N . “Mature” forms of polypeptides or proteins can also result from non-proteolytic post-translational modifications. Such proteolytic processes include, for example, glycosylation, myristylation, or phosphorylation. In general, a mature polypeptide or protein can result from only one manipulation of these processes, or a combination thereof.

  As used herein, “identical” residues correspond to those residues in a comparison between two sequences in which equal nucleotide bases or amino acid residues are the same residue in an alignment of the two sequences. Residues are instead described as “similar” or “positive” if the residues in the alignment where the comparison between the two sequences is at the same position in the comparison are the same or truncated amino acids described below.

  As used herein, “chemical composition” relates to a composition comprising at least one composition synthesized or extracted from a natural source. A chemical composition can be the product of a defined synthetic method. Such synthesized compositions have the properties defined in terms of molecular formulas, molecular structures related to the association of atoms attached to each, physical properties such as electropherographic or spectroscopic characterization, etc. herein. Understood in the book. Compositions extracted from natural sources provide a representation of defined properties, including molecular formulas, molecular structures related to the association of atoms attached to each, physical properties such as electropherography or spectroscopic characterization, etc. It is usefully analyzed by chemical and physical methods.

  As used herein, a “therapeutic reagent candidate” is a chemical composition comprising at least one substance that appears to bind to a target biopolymer. In important embodiments of the invention, the target biopolymer is a lipid such as a protein, polypeptide, nucleic acid, polysaccharide or proteoglycan, or complex lipid. The method of identifying a composition that binds to a target effectively eliminates a composition that has little or no binding affinity, and the potential for the identified chemical composition to have advantageous therapeutic applications. Increase. In the case where a “therapeutic reagent candidate” is a mixture of more than one chemical composition, subsequent screening methods are performed to identify specific substances that bind to the composition in the mixture and identify them as therapeutic reagent candidates. The

  As used herein, a “medicament” is a therapeutic reagent that uses a model of a disease state or pathology to identify candidates that have a desired therapeutic or beneficial therapeutic effect on the association with the disease or pathology. Provided by screening candidates. A candidate that successfully provides such an effect has been named herein a pharmaceutical product. Non-limiting examples of model systems used in such sorting include specific cell lines, cultured cells, tissue specimens, whole tissues, organ specimens, intact organs, and non-human mammals. Sorting utilizing at least one system, preferably more than one system, can be utilized to identify pharmaceuticals. The identified pharmaceutical can be explored in further studies with human subjects.

NOVX Nucleic Acids and Polypeptides NOVX Clones NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and situations. Various NOVX nucleic acids and polypeptides according to the present invention are useful as novel members of the protein family according to the presence of domains and sequences associated with the protein. In addition, NOVX nucleic acids and polypeptides can be used to identify proteins that are members of the family to which the NOVX polypeptide belongs.

  The NOVX genes and their corresponding encoded proteins are useful for preventing, treating or alleviating medical conditions, eg, by protein therapy or gene therapy. The pathology can be diagnosed by measuring the amount of the new protein in the sample or measuring the presence of the mutation in the new gene. Based on the tissues in which the NOVX gene is most highly expressed, the specific use for each NOVX gene will be described. Use includes developing products for diagnosing or treating various diseases and disorders.

  The NOVX nucleic acids and proteins of the present invention are useful as potential diagnostic and therapeutic applications and research tools. These include serving as specific or selective nucleic acids, or proteins, diagnostic markers and / or prognostic markers. Here, the presence or amount of nucleic acids or proteins, as well as potential therapeutic applications such as: (i) protein therapeutics, (ii) small molecule targets, (iii) antibody targets (therapeutic, Diagnostic, drug targeting / cytotoxic antibodies), (iv) nucleic acids useful for gene therapy (gene delivery / gene surgery), and (v) compositions that promote tissue regeneration in vitro and in vivo, (vi) Biological defense weapon.

  In one particular embodiment, the invention provides a mature form having an amino acid sequence selected from the group consisting of (a) SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539; (b) A mature mutant having an amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer of 1 to 129) and SEQ ID NO: 539, provided that any amino acid in the mature form is a mature sequence. 15% or less of the amino acid residues therein are changed to different amino acids under the condition that they are changed; (c) selected from the group consisting of SEQ ID NO: 2n (n is an integer of 1 to 129) and SEQ ID NO: 539 (D) a variant having an amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, but specified by the selected sequence any The amino acid is changed to a different amino acid under the condition that 15% or less of the amino acid residues in the sequence are changed; and e) an amino acid selected from the group consisting of any fragment of a) to d) An isolated polypeptide containing the sequence is included.

  In another specific embodiment, the invention provides a mature form having an amino acid sequence selected from (a) SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539; (b) SEQ ID NO: A mature variant having an amino acid sequence selected from the group consisting of 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, wherein any amino acid in the mature form of the selected sequence is matured A variant that has been changed to a different amino acid, provided that 15% or less of the amino acid residues in the sequence of the type are changed; (c) SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: An amino acid sequence selected from the group consisting of 539; (d) a variant of an amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, and selected In the array A variant wherein the amino acid identified is changed to a different amino acid, provided that 15% or less of the amino acid residues in the sequence are changed; and (e) SEQ ID NO: 2n (n is an integer from 1 to 129) And at least a portion of a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 539, or any amino acid of the selected sequence, provided that less than 10% of the amino acid residues in the sequence are altered A polypeptide comprising an amino acid sequence selected from the group consisting of: a fragment of a nucleic acid encoding any variant of a polypeptide that has been altered to a different amino acid; and (f) a complementary molecule of any of the nucleic acid molecules. An isolated nucleic acid molecule comprising a nucleic acid sequence to be

  In yet another particular embodiment, the present invention includes an isolated nucleic acid molecule. Wherein the nucleic acid molecule comprises (a) a nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538; (b) SEQ ID NO: 2n-1 (n Is an integer from 1 to 129) and one or more nucleotides in the nucleotide sequence selected from the group consisting of SEQ ID NO: 538 is selected from the group consisting of the selected sequence % A nucleotide sequence that has been changed to a different nucleotide under the condition that it is subject to change; (c) a sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 And (d) a nucleotide selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 Consisting of a nucleic acid fragment in which one or more nucleotides in the sequence are changed from those selected from the group consisting of selected sequences to different nucleotides provided that not more than 15% of the nucleotides are changed Contains a nucleotide sequence selected from the group.

  One aspect of the invention pertains to isolated nucleic acid molecules that encode NOVX polypeptides or biologically active portions thereof. The invention also includes nucleic acid fragments sufficient for use as hybridization probes to identify nucleic acids encoding NOVX (eg, of NOVX mRNA), and PCR for amplification and / or mutation of NOVX nucleic acid molecules. Includes fragments for use as primers. As used herein, the term “nucleic acid molecule” refers to a DNA molecule (eg, cDNA or genomic DNA), an RNA molecule (eg, mRNA), a DNA analog or RNA analog produced using nucleotide analogs, And their derivatives, fragments and homologues. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA.

  The NOVX nucleic acid can encode a mature NOVX polypeptide. As used herein, a “mature” form of a polypeptide or protein disclosed in the present invention is a naturally occurring polypeptide, or precursor form, or the product of a proprotein. Naturally occurring polypeptides, precursors or proproteins include, by way of non-limiting example, full-length gene products encoded by the corresponding genes. Alternatively, it may be defined as a polypeptide, precursor or proprotein encoded by the ORFs described herein. A “mature” form of a product also occurs, as a non-limiting example, as a result of one or more naturally occurring processing steps that can occur in a cell (host cell) that produces a gene product. Examples of such processing steps that lead to a “mature” form of a polypeptide or protein include cleavage of the N-terminal methionine residue encoded by the initiation codon of the ORF, or proteolytic cleavage of the signal peptide or leader sequence. Thus, a mature form resulting from a precursor polypeptide or protein having residues 1-N where residue 1 is the N-terminal methionine has residues 2-N remaining after removal of the N-terminal methionine. Alternatively, the mature form resulting from a precursor polypeptide or protein having residues 1-N that cleaves the N-terminal signal sequence from residues 1-residue M is derived from the remaining residues M + 1-residue N. Has a residue. Further, as used herein, a “mature” form of a polypeptide or protein may result from a post-translational modification step other than a proteolytic cleavage event. Such further methods include, but are not limited to glycosylation, myristoylation, or phosphorylation. In general, a mature polypeptide or protein can be produced by manipulation of only one of these methods, or any combination thereof.

  As used herein, the term “probe” preferably refers to a variable length nucleic acid sequence that is at least about 10 nucleotides (nt) to 100 nt, or depending on the particular use, for example, about 6,000 nt long. Point to. Probes may be used for detection of identical, similar or complementary nucleic acid sequences. Longer probes are generally obtained from natural or recombinant sources. Longer probes are slower to hybridize than oligomer probes of higher specificity and shorter length. Probes can be single-stranded or double-stranded and can be designed to have specificity in PCR, membrane-based hybridization techniques, or ELISA-like techniques.

  As used herein, the term “isolated” nucleic acid molecule is one that is separated from other nucleic acid molecules that are present in the natural source of the nucleic acid. Preferably, an “isolated” nucleic acid does not have sequences that naturally flank the nucleic acid in the genomic DNA of the organism from which the nucleic acid is derived (ie, sequences located at the 5 ′ and 3 ′ ends of the nucleic acid). For example, in various embodiments, the isolated NOVX nucleic acid molecule is about 5 kb, 4 kb naturally adjacent to the nucleic acid molecule in the genomic DNA of the cell / tissue from which the nucleic acid is derived (eg, brain, heart, liver, spleen, etc.). It may contain no more than 3 kb, 2 kb, 1 kb, 0.5 kb, or 0.1 kb nucleotide sequence. Furthermore, an “isolated” nucleic acid molecule, such as a cDNA molecule, may be substantially free of other cellular material, or media, or chemical precursors or other chemicals.

  The nucleic acid molecule of the present invention, for example, a nucleic acid molecule having a nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer of 1 to 129) and SEQ ID NO: 538, or a complement of this nucleotide sequence Can be isolated using standard molecular biology techniques and the sequence information provided herein. Standard hybridization and cloning techniques using all or part of a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 as a hybridization probe (E.g., Sambrook, et al., (Eds.), MOLECULAR CLONING: A LABORATORY MANUAL 2nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989; and Ausubel, et al., (Eds.), NOVX molecules can be isolated using CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, as described in John Wiley & Sons, New York, NY, 1993).

  The nucleic acids of the invention can be amplified with appropriate oligonucleotide primers according to standard PCR amplification techniques using cDNA, mRNA, or alternatively genomic DNA as a template. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. In addition, oligonucleotides corresponding to NOVX nucleotide sequences can be prepared by standard synthetic techniques, eg, using an automated DNA synthesizer.

  The term “oligonucleotide” as used herein refers to a series of linking nucleotide residues where the oligonucleotide has a sufficient number of nucleotide bases to be used in a PCR reaction. Short oligonucleotide sequences can be based on or designed from genomic or cDNA sequences. Short oligonucleotide sequences are then used to amplify, confirm, or reveal the presence of identical, similar, or complementary DNA or RNA in a particular cell or tissue. The oligonucleotide comprises a p portion of a nucleic acid sequence that is about 10 nt, 50 nt, or 100 nt in length, preferably about 15 nt to 30 nt in length. In one embodiment of the invention, the oligonucleotide comprising a nucleic acid molecule of less than 100 nt in length is a sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 It further comprises at least 6 consecutive nucleotides derived from, or complements thereof. Oligonucleotides can be chemically synthesized and used as probes.

  In another embodiment, the isolated nucleic acid molecule of the invention comprises the complementary sequence of a nucleotide sequence from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, or Includes a nucleic acid molecule that is the complement of a portion of a nucleotide sequence (eg, a fragment that can be used as a probe, primer, or fragment encoding a biologically active portion of a NOVX polypeptide). A nucleic acid molecule complementary to a nucleotide sequence from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 is SEQ ID NO: 2n-1 (n is an integer from 1 to 129) And nucleotides derived from the group consisting of SEQ ID NO: 538, which are sufficiently complementary to the nucleotide sequence from the group consisting of SEQ ID NO: 538 and SEQ ID NO: 538 It can hydrogen bond with the sequence with little or no mismatch. Therefore, the nucleic acid molecule forms a stable double strand.

  As used herein, the term “complementary” refers to Watson-Crick or Hoogsteen base pairing between nucleotide units of a nucleic acid molecule, and the term “binding” refers to two polypeptides or compounds, or related polypeptides. Or means a physical or chemical interaction between compounds or combinations thereof. Binding includes ionic, non-ionic, van der Waals, hydrophobic interactions, and the like. The physical interaction can be direct or indirect. Indirect interactions may be through or due to the action of other polypeptides or compounds. Direct binding refers to an interaction that does not occur through or due to the action of another polypeptide or compound but instead takes place without other substantial chemical mediators.

  As provided herein, “fragments” are at least 6 fragments that are long enough to allow specific hybridization in the case of nucleic acids, and long enough for specific recognition of epitopes in the case of amino acids. Defined as a sequence of (contiguous) nucleic acids or a sequence of at least 4 (contiguous) amino acids, and some part of most shorter than the full length. Fragments can be derived from any contiguous portion of the selected nucleic acid or amino acid sequence. Derivatives are nucleic acid or amino acid sequences generated either directly from the original compound, either by modification or by partial substitution. Analogs are nucleic acid or amino acid sequences that have a structure that is similar but not identical to the original compound, eg, they differ from a natural compound with respect to a particular component or side chain. Analogs can be synthetic, can be derived from different evolutionary origins, and can have similar or opposite metabolic activities compared to wild-type. A homologue is the nucleic acid or amino acid sequence of a particular gene derived from a different species.

  A full-length NOVX clone is identified as containing an ATG translation start codon and an in-frame stop codon. Any disclosed NOVX nucleotide sequence lacking the ATG initiation codon therefore encodes a truncated C-terminal fragment of each NOVX polypeptide and extends in the 5 'direction of the sequence disclosed by the corresponding full length cDNA. Require to do. Any disclosed NOVX nucleotide sequence lacking an in-frame stop codon similarly encodes a truncated N-terminal fragment of the respective NOVX polypeptide, such that the corresponding full-length cDNA extends in the 3 'direction of the disclosed sequence. Request.

  Derivatives and analogs can be full length or other than full length if the derivative or analog contains a modified nucleic acid or amino acid as described below. The nucleic acid or protein derivatives or analogs of the present invention, in various embodiments, at least in comparison to alignment sequences aligned over nucleic acid or amino acid sequences of the same size or aligned by computer homology programs known in the art. Molecules comprising regions that are substantially homologous to the nucleic acids or proteins of the invention with about 70%, 80%, or 95% identity (preferably 80-95% identity), or nucleic acids encoding them Include, but are not limited to, molecules that are capable of hybridizing under stringent, moderately stringent, or low stringency conditions to the complement of the sequence encoding the protein. See, for example, Ausubel, et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, NY, 1993 and the following.

  "Homologous nucleic acid sequence" or "homologous amino acid sequence" or variants thereof refer to sequences characterized by homology at the nucleotide or amino acid level as described above. Homologous nucleotide sequences include those sequences that encode isoforms of the NOVX polypeptide. Isoforms can be expressed in different tissues of the same organism as a result of, for example, alternative splicing of RNA. Alternatively, isoforms can be encoded by different genes. In the present invention, a homologous nucleotide sequence includes a nucleotide sequence encoding a non-human species NOVX polypeptide, including but not limited to vertebrates. Thus, for example, include frogs, mice, rats, rabbits, dogs, cats, cows, horses, and other organisms. Homologous nucleotide sequences also include, but are not limited to, naturally occurring allelic variants and mutants of the nucleotide sequences set forth herein. However, homologous nucleotide sequences do not include the exact nucleotide sequence encoding human NOVX protein. A homologous nucleic acid sequence includes a nucleic acid sequence encoding a conservative amino acid substitution (see below) in any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, and Polypeptides having NOVX biological activity are included. Various biological activities of the NOVX protein are described below.

  NOVX polypeptides are encoded by the open reading frame (“ORF”) of NOVX nucleic acids. The ORF corresponds to a nucleotide that can potentially be translated into a polypeptide. A series of nucleic acids containing the ORF is not interrupted by a stop codon. The ORF representing the sequence encoding the full-length protein begins with an ATG “start” codon and ends with one of the three “stop” codons, TAA, TAG or TGA. For the purposes of the present invention, the ORF can be any portion of the coding sequence that may or may not have a start codon, a stop codon, or both. In order to consider the ORF as a good candidate for encoding a genuine cellular protein, minimal requirements are often defined, for example a series of DNAs encoding 50 amino acids or more.

  Nucleotide sequences determined from cloning of the human NOVX gene are for use in identifying and / or cloning NOVX homologues from other cell types, eg, other tissues, and NOVX homologues from other vertebrates Allows creation of probes and primers to be designed. The probe / primer typically comprises a substantially purified oligonucleotide. The oligonucleotide has at least about 12, 25, 50, 100, 150, 200, 250, 300, any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, A nucleotide sequence of 350 or 400 consecutive sense strands; or exact to any one antisense strand nucleotide sequence of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 It typically includes a region of nucleotide sequence that hybridizes under conditions.

  Probes based on human NOVX nucleotide sequences can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In various embodiments, the probe further comprises a label group attached thereto, eg, the group can be a radioisotope, a fluorescent compound, an enzyme, or a cofactor of the enzyme. Such probes are part of a diagnostic test kit for identifying cells or tissues that misexpress a NOVX protein, such as by measuring the level of a NOVX-encoding nucleic acid in a cell sample from a subject. For example, NOVX mRNA is detected or whether the genomic NOVX gene has been mutated or deleted.

  A “polypeptide having a biologically active portion of a NOVX polypeptide” includes mature forms as measured in a particular biological analysis and is similar, but not necessarily identical, to the activity of a polypeptide of the invention. It refers to a polypeptide that exerts no activity with or without dose dependency. A nucleic acid fragment encoding a “biologically active portion of NOVX” is a sequence of SEQ ID NOs: 2n-1 (where n is A portion of any one of the group consisting of SEQ ID NO: 538 and expressing the NOVX protein-encoding portion (eg, by in vitro recombinant expression), and the NOVX-encoding portion Can be prepared by evaluating the activity.

NOVX Nucleic Acid and Polypeptide Variants The present invention further provides the nucleotide sequence shown in any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 due to the degeneracy of the genetic code Thus encoding the same NOVX protein as encoded by the nucleotide sequence set forth in SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and any one of the group consisting of SEQ ID NO: 538 Includes molecules. In another embodiment, an isolated nucleic acid molecule of the invention has an amino acid sequence set forth in any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 It has a nucleotide sequence that encodes a protein.

  In addition to the human NOVX nucleotide sequence shown in any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, a DNA sequence that causes a change in the amino acid sequence of the NOVX polypeptide One of ordinary skill in the art will understand that multiple polymorphisms may exist in a population (eg, a human population). Such genetic polymorphisms in the NOVX gene may exist among individuals in the population due to natural allelic variation. The terms “gene” and “recombinant gene” as used herein refer to a nucleic acid molecule comprising an open reading frame (ORF) encoding a NOVX protein, preferably a vertebrate NOVX protein. Such natural allelic variations typically result in 1-5% variation in the nucleotide sequence of the NOVX gene. Any and all such nucleotide variations that are the result of natural allelic variation and do not alter the functional activity of the NOVX polypeptide and the amino acid polymorphisms in the NOVX polypeptide obtained therefrom are intended to be within the scope of the invention. .

  Furthermore, it has a nucleic acid molecule encoding a NOVX protein from another species, and thus a nucleotide sequence different from any of the group consisting of human SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 Nucleic acid molecules are intended to be within the scope of the present invention. Nucleic acid molecules corresponding to natural allelic variants and homologues of the NOVX cDNA of the present invention are obtained using human cDNA or a portion thereof as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions. It can be isolated based on homology with the human NOVX nucleic acids disclosed herein.

  Thus, in another embodiment, an isolated nucleic acid molecule of the invention is at least 6 nucleotides in length and is any member of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538. Hybridizes under stringent conditions to a nucleic acid molecule containing a single nucleotide sequence. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 750, 1000, 1500, or 2000 or more nucleotides in length. In yet another embodiment, the isolated nucleic acid molecule of the invention hybridizes to the coding region. As used herein, the term “hybridizes under stringent conditions” describes hybridization and washing conditions in which nucleotide sequences that are at least about 60% homologous to each other typically remain hybridized to each other. Intended.

  Homologues (i.e., nucleic acids encoding NOVX proteins from non-human species) or other related sequences (e.g., paralogs) can be converted to specific human sequences using methods well known in the art of nucleic acid hybridization and cloning. All or part of the probe is used as a probe, and can be obtained by low, medium, or high stringent hybridization.

  As used herein, the term “stringent hybridization conditions” refers to conditions under which a probe, primer or oligonucleotide hybridizes to a target sequence but not to other sequences. The exact conditions are sequence dependent and there are differences in different environments. Longer sequences hybridize specifically at higher temperatures than shorter sequences. In general, stringent conditions are selected to be about 5 ° C. lower than the melting temperature (Tm) for the specific sequence at a constant ionic strength and pH. Tm is the temperature (under a defined ionic strength, pH and nucleic acid concentration) at which 50% of the probe complementary to the target sequence hybridizes to the target sequence at equilibrium. Since the target sequence is generally present in excess at Tm, it accounts for 50% of the probe at equilibrium. Typically, the strict conditions are pH 7.0-8.3 and a salt concentration of less than about 1.0M sodium ion, typically about 0.01-1.0M sodium ion (or other salt), The temperature is at least about 30 ° C. for short probes, primers or oligonucleotides (eg, 10 nt to 50 nt) and at least about 60 ° C. for longer probes, primers and oligonucleotides. Stringent conditions can also be achieved with the addition of destabilizing agents such as formamide.

  The exact conditions are known to those skilled in the art and can be found in Ausubel, et al., (Eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY (1989), 6.3.1-6.3.6. it can. Preferably, the conditions are such that sequences that are at least 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other. is there. Non-limiting examples of stringent hybridization include 6 × SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA and 500 mg / ml denaturation. Hybridization is performed at 65 ° C. in a high salt buffer containing salmon sperm DNA, and then washed once or more at 50 ° C. in 0.2 × SSC, 0.01% BSA. An isolated nucleic acid molecule of the invention that hybridizes under stringent conditions to any sequence of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 is naturally occurring Corresponds to a nucleic acid molecule. As used herein, the term “naturally-occurring” nucleic acid molecule refers to an RNA or DNA molecule having a nucleic acid sequence that occurs in nature (eg, encodes a natural protein).

  In a second embodiment, a nucleic acid molecule comprising any one nucleotide sequence of the group consisting of SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and SEQ ID NO: 538, or a fragment, analog or Nucleic acid sequences that hybridize to the derivatives under moderately stringent conditions are provided. Non-limiting examples of moderately stringent hybridization conditions include hybridization at 55 ° C. in 6 × SSC, 5 × Denhardt's solution, 0.5% SDS and 100 mg / ml denatured salmon sperm DNA, followed by 1 × Rinse one or more times at 37 ° C. in SSC, 0.1% SDS. Other moderately stringent conditions that can be used are well known in the art. See, for example, Ausubel, et al. (Eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Krieger, 1990; GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY. .

  In a third embodiment, a nucleic acid molecule comprising any one nucleotide sequence of the group consisting of SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and SEQ ID NO: 538, or a fragment, analog or Nucleic acid sequences that hybridize to derivatives under low stringency conditions are provided. Non-limiting examples of low stringency hybridization conditions include 35% formamide, 5 × SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.0%. Hybridization in 2% BSA, 100 mg / ml denatured salmon sperm DNA, 10% (wt / vol) dextran sulfate at 40 ° C., followed by 2 × SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, And washing once or more at 50 ° C. in 0.1% SDS. Other less stringent conditions that can be used (eg, used for interspecies hybridization) are well known in the art. For example, Ausubel, et al. (Eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990; GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY .; Shilo and Weinberg 1981. Proc Natl Acad Sci USA 78: 6789-6792.

Conservative variation In addition to the naturally occurring allelic variation of the NOVX sequence that may be present in the population, any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 Those skilled in the art further understand that mutations can be introduced into one nucleotide sequence by mutation, thereby leading to changes in the amino acid sequence of the encoding NOVX protein without altering the functional activity of the NOVX protein. For example, nucleotide substitutions leading to amino acid substitutions at “non-essential” amino acid residues may be made in any one sequence of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539. “Non-essential” amino acid residues are those residues that can change the wild-type sequence of the NOVX protein without altering their biological activity, while “essential” amino acid residues are such biological activity. Need to. For example, amino acid residues that are conserved in the NOVX protein of the present invention are not expected to be particularly resistant to conversion. Amino acids for which conservative substitutions can be made are well known in the art.

  Another aspect of the invention pertains to nucleic acid molecules encoding NOVX proteins that contain changes in amino acid residues that are not essential for activity. Such NOVX protein differs in amino acid sequence from any of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, but still retains biological activity. In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein is any one of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539. An amino acid sequence that is at least about 45% homologous to one amino acid sequence. Preferably, the protein encoded by the nucleic acid molecule is at least about 70% homologous to any one of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539; SEQ ID NO: 2n (n is an integer from 1 to 129) and at least about 80% homologous to any one of the group consisting of SEQ ID NO: 539; even more preferably, SEQ ID NO: 2n (n is 1 to 1) 129) and at least about 90% homologous to any one of the group consisting of SEQ ID NO: 539; most preferably from SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539 Is at least about 95% homologous to any one of the groups.

  An isolated nucleic acid molecule that encodes a NOVX protein that is homologous to any one protein of the group consisting of SEQ ID NO: 2n (where n is an integer from 1 to 129) and SEQ ID NO: 539 is one or more amino acids 1 in any one nucleotide sequence of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 for introduction into a protein encoding a substitution, addition or deletion It can be created by introducing one or more nucleotide substitutions, additions or deletions.

  Mutations are made in any one of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, such as site-directed mutagenesis and PCR-mediated mutagenesis. It can be introduced by standard techniques. Preferably, conservative amino acid substitutions are made at one or more predicted non-essential amino acid residues. A “conservative amino acid substitution” is one of the amino acid residues that is replaced with an amino acid residue having a similar side chain. A family of amino acid residues with similar side chains has been defined within the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), amino acids with acidic side chains (e.g., aspartic acid, glutamic acid), amino acids with uncharged side chains (e.g., glycine, Asparagine, glutamine, serine, threonine, tyrosine, cysteine), amino acids with nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), amino acids with beta side chains (e.g., Threonine, valine, isoleucine), as well as amino acids with aromatic side chains (eg tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted nonessential amino acid residue in a NOVX protein is replaced with another amino acid residue from the same side chain family. Alternatively, in another embodiment, mutations can be introduced randomly along all or part of a NOVX coding sequence, such as saturation mutagenesis, and the resulting mutations The body can be screened for NOVX biological activity to identify mutants that retain activity. Following mutagenesis of any of the group consisting of SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and SEQ ID NO: 538, the encoded protein is obtained by any recombinant technique known in the art. Expressed and protein activity can be measured.

  Amino acid family relationships can also be determined based on side chain interactions. Substituted amino acids can be fully conserved “strong” residues or fully conserved “weak” residues. The “strong” group of conserved amino acid residues can be any one of the following groups: STA, NEQK, NHQK, NDEQ, QHRK, MILV, MILF, HY, FYW, where the one letter code for amino acids Are grouped with amino acids that can be substituted for each other. Similarly, the “weak” group of conserved residues can be any one of the following: CSA, ATV, SAG, STNK, STPA, SGND, SNDEKK, NDEQHK, NEQHRK, HFY, where each Group letters represent the single letter code for amino acids.

In one embodiment, the mutant NOVX protein is (i) a protein that interacts with other NOVX proteins, other cell surface proteins or their biologically active sites: an activity that forms a protein, (ii) a mutant NOVX protein May be assayed for complex formation between certain NOVX ligands, or (iii) the activity of mutant NOVX proteins that bind to intracellular target proteins or their biologically active sites (eg, avidin protein).
In yet another embodiment, mutant NOVX proteins can be assayed for activity that modulates a specific biological function (eg, modulation of insulin secretion).

Antisense Nucleic Acid Another aspect of the present invention is a nucleic acid molecule comprising any one nucleotide sequence of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538, or thereof It relates to an isolated antisense nucleic acid molecule capable of hybridising or complementary to a fragment, homologue or derivative. An “antisense” nucleic acid comprises a nucleotide sequence that is complementary to a “sense” nucleic acid encoding a protein (eg, complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence). In particular embodiments, antisense nucleic acid molecules comprising at least about 10, 25, 50, 100, 250, or 500 nucleotides or a sequence complementary to the entire NOVX coding strand, or only a portion thereof, are provided. A nucleic acid molecule encoding a fragment, homologue, derivative and analogue of any one NOVX protein of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, SEQ ID NO: 2n (n Is an integer from 1 to 129) and any one of the group consisting of SEQ ID NO: 538 is provided in addition to a complementary antisense nucleic acid.

  In one embodiment, the antisense nucleic acid molecule is antisense to a “coding region” of the coding strand of a nucleotide sequence encoding a NOVX protein. The term “coding region” refers to a region of a nucleotide sequence that includes codons translated into amino acid residues. In another embodiment, the antisense nucleic acid molecule is antisense to a “non-coding region” of the coding strand of a nucleotide sequence encoding a NOVX protein. The term “noncoding region” refers to 5 ′ and 3 ′ sequences that flank the coding region that are not translated into amino acids (ie, also referred to as 5 ′ untranslated region and 3 ′ untranslated region).

  Given the sequence of the coding strand encoding the NOVX protein disclosed herein, the antisense nucleic acids of the invention can be designed according to the rules of Watson and Crick or Hoogsteen base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of NOVX mRNA, but more preferably is an oligonucleotide that is antisense to only a portion of the coding or non-coding region of NOVX mRNA. . For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of NOVX mRNA. Antisense oligonucleotides can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides in length. An antisense nucleic acid of the invention can be constructed using chemical synthesis or enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) increases the biological stability of a naturally occurring nucleotide or molecule, or a double-stranded physical stability that forms between an antisense nucleic acid and a sense nucleic acid. Various modified nucleotides designed to increase can be chemically synthesized (eg, phosphorothioate derivatives and nucleotides substituted with acridine can be used).

  Examples of modified nucleotides that can be used to make antisense nucleic acids include: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5 -(Carboxyhydroxymethyl) uracil, 5-carboxymethylaminomethyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosilk enosine, inosine, N6-isopentenyladenine, 1-methylguanine 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5 -Methoxy Minomethyl-2-thiouracil, beta-D-mannosilk eosin, 5′-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wivetoxosin, pseudouracil , Queosin, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 5-methyl 2-thiouracil, 3- (3-amino-3-N-2-carboxypropyl) uracil, (acp3) w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid is subcloned in the antisense orientation of the nucleic acid (ie, RNA transcribed from the inserted nucleic acid is the antisense orientation for the target nucleic acid of interest and is further described in the subsection below). Can be produced biologically using the expression vector.

  Protein expression by hybridizing or binding the antisense nucleic acid molecules of the present invention to cellular mRNA and / or genomic DNA that encodes certain NOVX proteins thereby (eg, inhibiting transcription and / or translation) Is typically administered to a subject so as to inhibit or made in tissue. Hybridization can also be due to the complementarity of conventional nucleotides to form stable duplexes. Alternatively, for example, in the case of an antisense nucleic acid that binds to a DNA double strand, it can be through a specific interaction in the double-stranded major groove. An example of a route of administration of antisense nucleic acid molecules of the invention includes direct injection into a tissue site. Alternatively, antisense nucleic acid molecules can be modified to selected cellular targets where they can be administered systemically. For example, for systemic administration, antisense molecules are specifically bound to receptors or antigens expressed on selected cell surfaces (e.g., linking antisense nucleic acid molecules to peptides or cell surfaces). May be modified (by antibodies that bind to receptors or antigens). Antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. In order to deliver sufficient nucleic acid molecules, a vector structure that places the antisense nucleic acid molecule under the control of a strong pol II or pol III promoter is preferred.

  In yet another embodiment, the antisense nucleic acid molecule of the invention is an α-anomeric nucleic acid molecule. α-anomeric nucleic acid molecules form specific double-stranded hybrids with complementary RNAs, which are opposite to normal β units and run parallel to each other. See, for example, Gaultier, et al., 1987. Nucl. Acids Res. 15: 6625-6641. Antisense nucleic acid molecules can also be 2′-o-methyl ribonucleotides (see, eg, Inoue, et al. 1987. Nucl. Acids Res. 15: 6131-6148) or chimeric RNA-DNA analogs (eg, Inoue, et al., 1987. FEBS Lett. 215: 327-330).

Ribozymes and PNA moieties Nucleic acid modifications include, by way of non-limiting example, modified bases and nucleic acids modified or derived from sugar phosphate backbones. These modifications are made at least in part to enhance the chemical stability of the modified nucleic acid so that it can be used, for example, as an antisense that binds to the nucleic acid in therapeutic applications to a subject.

  In one embodiment, the antisense nucleic acid of the invention is a ribozyme. Ribozymes are catalytically active RNA molecules with ribonuclease activity that are capable of cleaving single-stranded nucleic acids such as mRNA having a complementary region. Thus, ribozymes (eg, hammerhead ribozymes as described in Haselhoff and Gerlach 1988. Nature 334: 585-591) are used to catalytically cleave NOVX mRNA transcripts, thereby NOVX mRNA. Translation may be inhibited. A ribosome having specificity for a nucleic acid encoding NOVX is a group consisting of the nucleotide sequence of one NOVX cDNA disclosed herein (ie, SEQ ID NO: 2n-1 (n is an integer from 1 to 129)) and SEQ ID NO: 538. Can be designed based on any one of For example, a derivative of Tetrahymena L-19IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to a nucleotide sequence that can be cleaved in mRNA encoding NOVX. See, for example, Cech, et al., US Pat. No. 4,987,071, and Cech, et al., US Pat. No. 5,116,742. NOVX mRNA can also be used to select catalytic RNAs with specific ribonuclease activity from a pool of RNA molecules. See, for example, Bartel et al., (1993) Science 261: 1411-1418.

  On the other hand, NOVX gene expression is inhibited by targeting a nucleotide sequence complementary to a regulatory region of NOVX nucleic acid (eg, NOVX promoter and / or enhancer), and triple helical that blocks transcription of NOVX gene in the target cell. A structure can be formed. See, for example, Helene, 1991. Anticancer Drug Des. 6: 569-84; Helene, et al. 1992. Ann. NY Acad. Sci. 660: 27-36; Maher, 1992. Bioassays 14: 807-15. .

  In various embodiments, NOVX nucleic acids can be modified at the base moiety, sugar moiety, or phosphate backbone moiety to improve, for example, molecular stability, hybridization, or solubility. For example, the nucleic acid deoxyribose phosphate backbone can be modified to produce peptide nucleic acids. See, for example, Hyrup, et al., 1996. Bioorg Med Chem 4: 5-23. As used herein, the term “peptide nucleic acid” or “PNA” refers to a nucleic acid mimetic (eg, a DNA mimetic) that replaces the deoxyribose phosphate backbone with a pseudopeptide backbone and retains only four nucleotide bases. . The neutral backbone of PNA is shown to allow specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers is described in Hyrup, et al., 1996. supra; Perry-O'Keefe, et al., 1996. Proc. Natl. Acad. Sci. USA 93: 14670-14675 It can be performed using a standard solid phase peptide synthesis protocol.

NOVX PNA may be used for therapeutic and diagnostic applications. For example, PNA can be used as an antisense or antigenic agent for sequence specific modulation of gene expression, for example by inducing transcription or translational arrest or inhibiting replication. NOVX PNA can also be used, for example, as an artificial restriction enzyme that can be used in combination with single base pair mutations in genes (eg, PNA directing PCR fixation; other enzymes such as S ₁ nuclease) ( Hyrup, et al., 1996.supra); or as DNA sequence and hybridization probes or primers (Hyrup, et al., 1996, supra; see Perry-O'Keefe, et al., 1996. supra) )) Can also be used.

  In another embodiment, the NOVX PNA is modified, eg, by attaching a lipophilic or other auxiliary group to the PNA, by generation of a PNA-DNA chimera, or by liposomes or the art Can be used to enhance their stability or cellular uptake by use of other drug delivery techniques known in the art. For example, a PNA-DNA chimera of NOVX can be generated that can combine the beneficial properties of PNA and DNA. Such chimeras allow DNA recognition enzymes (eg, RNase H and DNA polymerase) to interact with the DNA moiety, while the PNA moiety provides binding high affinity and binding specificity. PNA-DNA chimeras can be joined using linkers of appropriate lengths selected for base linkage, number of linkages between nucleotide bases and orientation (see Hyrup, et al., 1996. supra). Synthesis of PNA-DNA chimeras can be performed as described in Hyrup, et al., 1996. supra and Finn, et al., 1996. Nucl Acids Res 24: 3357-3363. For example, a DNA strand can be synthesized on a solid support using standard phosphoramidite coupling chemistry. Modified nucleoside analogs, such as 5 ′-(4-methoxytrityl) amino-5′-deoxythymidine phosphoramidites, can then be used between the PNA and the 5 ′ end of the DNA. See, for example, Mag, et al., 1989. Nucl Acid Res 17: 5973-5988. The PNA monomer is then coupled stepwise to produce a chimeric molecule having a 5 ′ PNA segment and a 3 ′ DNA segment. See, for example, Finn, et al., 1996. supra. Alternatively, the chimeric molecule can be synthesized with a 5 ′ DNA segment and a 3 ′ PNA segment. See, for example, Petersen, et al., 1975. Bioorg. Med. Chem. Lett. 5: 1119-11124.

  In other embodiments, the oligonucleotides are attached to groups such as peptides (e.g., to target host cell receptors in vivo), or cell membranes (e.g., Letsinger, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 6553-6556; Lemaitre, et al., 1987. Proc. Natl. Acad. Sci. 84: 648-652; Agents that facilitate transport across (see WO89 / 10134). In addition, oligonucleotides can be combined with hybridization-induced cleavage agents (see, eg, Krol, et al., 1988. BioTechniques 6: 958-976) or intercalating agents (eg, Zon, 1988. Pharm. Res. 5: 539 -549). For this purpose, oligonucleotides can be conjugated to, for example, peptides, hybridization-inducing cross-linking agents, transport agents, hybridization-inducing cleavage agents, and the like.

NOVX polypeptides Polypeptides according to the present invention include polypeptides that contain the amino acid sequence of a NOVX polypeptide provided in the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539. The present invention can also convert any residue thereof from the corresponding residue shown in the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, while its NOVX activity and It includes mutated or variant proteins that still encode proteins that retain physiological function, or functional fragments thereof.

  In general, one NOVX variant that retains a NOVX-like function includes any variant that substitutes a residue at a particular site in the sequence with another amino acid, with another residue between two residues of the parent protein. Or further including the possibility of inserting multiple residues as well as the possibility of deleting one or more residues from the parent sequence. Any amino acid substitution, insertion or deletion is encompassed by the present invention. In preferred circumstances, the substitution is a conservative substitution as defined above.

  One aspect of the present invention relates to isolated NOVX proteins and biologically active portions thereof, or derivatives, fragments, analogs or homologues thereof. Polypeptide fragments suitable for use as an immunogen for the production of anti-NOVX antibodies can also be provided. In one embodiment, native NOVX protein can be isolated from a cell or tissue source by a suitable purification scheme using standard protein purification techniques. In another embodiment, NOVX protein is produced by recombinant DNA technology. As an alternative to recombinant expression, certain NOVX proteins or polypeptides can be chemically synthesized using standard peptide synthesis techniques.

  An “isolated” or “purified” polypeptide or protein or biologically active portion thereof is substantially free of cellular or tissue source cellular material or other contaminating protein from which the NOVX protein is derived, or chemically When synthesized, it is substantially free of chemical precursors or other chemicals. The term “substantially free of cellular material” includes a sample of NOVX protein that has separated the protein from the cellular components of the original cell from which the protein was isolated or recombinantly produced. In one embodiment, the term “substantially free of cell source” means less than about 30% (dry weight ratio) of non-NOVX protein (also referred to herein as “contaminating protein”), more preferably about Samples of NOVX protein having less than 20% non-NOVX protein, even more preferably less than about 10% non-NOVX protein, and most preferably less than about 5% non-NOVX protein are included. In recombinantly producing NOVX proteins or biologically active portions thereof, it is also preferably substantially free of medium, ie, the medium is less than about 20% of the volume of the NOVX protein specimen, more preferably It represents less than about 10%, and most preferably less than 5%.

  The term “substantially free of chemical precursors or other chemicals” includes a sample of NOVX protein where proteins are separated from chemical precursors or other chemicals involved in protein synthesis. To do. In one embodiment, the term “substantially free of chemical precursors or other chemicals” refers to less than about 30% (dry weight ratio) chemical precursors or non-NOVX chemicals, more preferably Less than about 20% chemical precursor or non-NOVX chemical, even more preferably less than about 10% chemical precursor or non-NOVX chemical, and most preferably less than about 5% chemical precursor or non-NOVX Includes specimens of NOVX protein with chemicals.

The biologically active portion of the NOVX protein comprises the amino acid sequence of the NOVX protein that contains fewer amino acids than the full-length NOVX protein and exhibits at least one activity of the NOVX protein (eg, SEQ ID NO: 2n, where n is an integer from 1 to 129). And a fully homologous amino acid sequence shown in any one of the group consisting of SEQ ID NO: 539 or an amino acid sequence shown in the amino acid sequence of NOVX protein). Typically, the biologically active portion includes a domain or motif having at least one activity of the NOVX protein. A biologically active portion of a NOVX protein can be a polypeptide, eg, 10, 25, 50, 100 or more amino acid residues long.
In addition, other biologically active portions lacking other regions of the protein can be prepared by recombinant techniques and evaluated for one or more of the functional activities of the native NOVX protein.

  In one embodiment, the NOVX protein has an amino acid sequence set forth in any one of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539. In another embodiment, the NOVX protein is substantially homologous to any one of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, and SEQ ID NO: 2n (n is 1) and the functional activity of any one protein of the group consisting of SEQ ID NO: 539 but still as detailed below due to natural allelic variants or mutations Make different. Thus, in another embodiment, the NOVX protein comprises an amino acid sequence that is at least about 45% homologous to any one amino acid sequence of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539 A protein that retains the functional activity of any one NOVX protein of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539.

Measuring homology between two or more sequences. To determine the percentage of homology between two amino acid sequences or two nucleic acids, the sequences are aligned for optimal comparison purposes (eg, gaps, Can be placed within the first amino acid or nucleic acid sequence for optimal alignment with the second amino acid or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid or nucleic acid sites are then compared. When a site in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding site in the second sequence, both molecules are homologous at that site (ie, as used herein, amino acids or Nucleic acid “homology” is equivalent to amino acid or nucleic acid “identity”).

  Nucleic acid sequence homology can be measured as the degree of identity between two sequences. Homology can be measured using computer programs known in the art, such as GAP software provided in the GCG program package. See Needleman and Wunsch, 1970. J Mol Biol 48: 443-453. Using the GCG GAP software with the following settings: GAP generation penalty of 5.0 or GAP extension penalty of 0.3, the coding region of the above-mentioned similar nucleic acid sequence is SEQ ID NO: 2n-1 (n is 1 to 129) And preferably at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99 and the CDS (encoding) portion of the DNA sequence from the group consisting of SEQ ID NO: 538 % Degree of identity.

  The term “sequence identity” refers to the extent to which two polynucleotide or polypeptide sequences are identical from residue to residue over a particular region of comparison. The term “percent of sequence identity” is used to compare two sequences that are optimally aligned over the region of comparison and to determine the number of matching sites (for example, A, in the case of nucleic acids, A, The number of sites where T, C, G, U, or I) is present in both sequences is determined, the number of matching sites is divided by the total number of sites in the comparison region (ie, window size), and the quotient is 100. Can be calculated by multiplying to obtain a percentage of sequence identity. As used herein, the term “substantial identity” means a property of a polynucleotide sequence. Here, the polynucleotide comprises at least 80% sequence identity, preferably at least 85% sequence identity, and often 90-95% sequence identity, more usually compared to the standard sequence over the comparison region. Includes sequences with at least 99% sequence identity.

Chimeric or fusion protein The present invention also provides a NOVX chimeric or fusion protein. As used herein, certain NOVX “chimeric proteins” or “fusion proteins” include certain NOVX polypeptides operably linked to non-NOVX polypeptides. “NOVX polypeptide” refers to a polypeptide having an amino acid sequence corresponding to one NOVX protein of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539, while “non- “NOVX polypeptide” refers to a polypeptide having an amino acid sequence corresponding to a protein that is not substantially homologous to the NOVX protein, eg, a protein that is from the same or different organism, unlike the NOVX protein. Within a NOVX fusion protein, a NOVX polypeptide may correspond to all or part of a NOVX protein. In one embodiment, a NOVX fusion protein includes at least one biologically active portion of a NOVX protein. In another embodiment, a NOVX fusion protein comprises at least two biologically active portions of a NOVX protein. In yet another embodiment, a NOVX fusion protein comprises at least three biologically active portions of a NOVX protein. Within the fusion protein, the term “operably linked” shall indicate that the NOVX polypeptide and the non-NOVX polypeptide are fused in-frame to each other. The non-NOVX polypeptide can be fused to the N-terminus or C-terminus of the NOVX polypeptide.

  In one embodiment, the fusion protein is a GST-NOVX fusion protein. Here, the NOVX sequence is fused to the C-terminus of GST (glutathione S transferase). Such fusion proteins can facilitate the purification of recombinant NOVX polypeptides.

  In another embodiment, the fusion protein is a NOVX protein containing a heterologous signal sequence at its N-terminus. In certain host cells (eg, mammalian host cells), NOVX expression and / or secretion may be enhanced through the use of heterologous signal sequences.

  In yet another embodiment, the fusion protein is a NOVX-immunoglobulin fusion protein. Here, the NOVX sequence is fused to a sequence derived from a member of the immunoglobulin protein family. A NOVX-immunoglobulin fusion protein of the invention is incorporated into a pharmaceutical composition and administered to a subject to inhibit the interaction of a NOVX ligand with a NOVX protein on the cell, thereby inhibiting NOVX-mediated in vivo. Information transmission can be suppressed. NOVX-immunoglobulin fusion proteins can be used to affect the bioavailability of certain NOVX-like ligands. Inhibition of the NOVX ligand / NOVX interaction may be therapeutically useful for the treatment of proliferation and differentiation disorders and for modulating (eg, enhancing or inhibiting) cell survival. In addition, the NOVX-immunoglobulin fusion proteins of the present invention are used to produce anti-NOVX antibodies in a subject, purify NOVX ligand, and inhibit the interaction of NOVX with a NOVX ligand in screening assays Can be identified.

  The NOVX chimeric or fusion proteins of the invention can be produced by standard recombinant DNA techniques. For example, DNA fragments encoding different polypeptide sequences can be used, for example, using blunted or staggered ends for ligation, cleavage with restriction enzymes to provide appropriate ends, appropriately filling sticky ends, desirably Ligated together in-frame according to conventional methods such as alkaline phosphatase treatment to prevent unbound and enzymatic coupling. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments is performed using anchor primers that produce complementary overhangs between two consecutive gene fragments, which are subsequently annealed and reamplified to produce a chimeric gene sequence. (See, eg, Ausubel, et al. (Eds.) CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, 1992). Moreover, many expression vectors (eg, GST polypeptides) that already encode a fusion moiety are commercially available. A nucleic acid encoding NOVX can be cloned into such an expression vector such that the fusion moiety binds in-frame to the NOVX protein.

NOVX Agonists and Antagonists The present invention also includes variants of the NOVX protein that function as either NOVX agonists (ie, mimetics) or NOVX antagonists. NOVX protein variants may be generated by mutation (eg, point mutation or truncation of the NOVX protein). NOVX protein agonists retain substantially the same biological activity or subset thereof as the naturally occurring form of NOVX protein. NOVX protein agonists inhibit the activity of one or more naturally occurring forms of NOVX protein by, for example, antagonistic binding to downstream or upstream members of the cellular signaling cascade, including NOVX protein Can do. Thus, specific biological effects can be exerted by treatment with variants having limited functions. In one embodiment, treatment of a subject with a variant having a subset of the biological activity of a naturally occurring form of the protein has fewer side effects in the subject than treatment with a naturally occurring form of NOVX protein.

  Variants of NOVX proteins that function as either NOVX agonists (ie, mimetics) or NOVX antagonists screen recombinant libraries of NOVX protein mutants (eg, truncation mutants) for NOVX protein agonist or antagonist activity Can be identified. In one embodiment, the NOVX variant rich library is generated by recombinant (combinatorial) mutagenesis at the nucleic acid level and is encoded by the gene library rich in change. A library enriched in NOVX variants can be used, for example, to enzymatically link a mixture of synthetic oligonucleotides to a gene sequence so that a degenerate set of potential NOVX sequences can be expressed as individual polypeptides. Or alternatively as a set of larger fusion proteins (eg, for phage display) containing a set of NOVX sequences therein. There are a variety of methods that can be generated from a degenerate oligonucleotide sequence using a library of potential NOVX variants. Chemical synthesis of degenerate gene sequences can be performed with an automated DNA synthesizer, and the synthetic gene can then be ligated into an appropriate expression vector. The use of a degenerate set of genes allows the provision of a single mixture of all of the sequences that encode the desired set of potential NOVX sequences. Methods for synthesizing degenerate oligonucleotides are well known in the art. For example, Narang, 1983. Tetrahedron 39: 3; Itakura, et al., 1984. Annu. Rev. Biochem. 53: 323; Itakura, et al., 1984. Science 198: 1056; Ike, et al., 1983. See Nucl. Acids Res. 11: 477.

Polypeptide Library In addition, a fragment library of NOVX protein-encoding sequences can be used to generate a population enriched in NOVX fragments for screening and subsequent selection of NOVX protein variants. In one embodiment, a library of coding sequence fragments is subjected to nuclease treatment of a double stranded PCR fragment of a NOVX coding sequence under conditions where nicking occurs only once per molecule to denature the double stranded DNA. Forming a double-stranded DNA that can contain sense / antisense pairs from different nicking products, removing the single-stranded portion from the double helix regenerated by treatment with S ₁ nuclease, and the resulting fragment live Can be generated by linking the library to an expression vector. By this method, an expression library encoding various sized N-terminal or internal fragments of the NOVX protein can be derived.

  Various techniques are known in the art for screening gene products of recombinant (combinatorial) libraries generated by point mutations or truncations, and screening cDNA libraries for gene products having selected properties. . Such techniques are applicable to rapid screening of gene libraries generated by recombinant (combinatorial) mutagenesis of NOVX proteins. The most widely used technique applicable to high-throughput analysis to screen large gene libraries is to clone the gene library into a replicable expression vector and transform the appropriate cells with the resulting vector library, And the detection of the desired activity typically involves expressing the recombinant (combinatorial) gene under conditions that facilitate the isolation of the vector encoding the gene that detects the product. A new technique that increases the frequency of functional mutations in the library, repetitive ensemble mutagenesis (REM), can be used in combination with screening assays to identify NOVX mutants. See, for example, Arkin and Yourvan, 1992. Proc. Natl. Acad. Sci. USA 89: 7811-7815; Delgrave, et al., 1993. Protein Engineering 6: 327-331.

NOVX Antibody As used herein, the term “antibody” refers to an immunologically active portion of an immunoglobulin molecule and an immunoglobulin (Ig) molecule, ie, specifically binds (and immunologically reacts with) an antigen. ) Refers to a molecule containing an antigen binding site. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, F _ab , F _{ab ′} and F _{(ab ′) 2} fragments and a F _ab expression library. In general, antibody molecules obtained from humans relate to any of the classes IgG, IgM, IgA, IgE and IgD, which differ from each other due to the nature of the heavy chain present in the molecule. Certain classes have subclasses as well, such as IgG ₁ , IgG ₂ and others. Furthermore, in humans, the L chain can be a k chain or a lambda chain. Reference herein to antibodies includes a reference to all such classes, subclasses and types of human antibody species.

  Generate isolated antibodies immunospecifically using standard techniques for polyclonal and monoclonal antibody preparation using the isolated protein of the present invention, or a portion or fragment thereof, intended for use as an antigen, as an immunogen Can do. The full-length protein may be used, or alternatively, the present invention provides an antigenic peptide fragment of an antigen for use as an immunogen. The antigenic peptide fragment comprises at least six amino acid sequences of the full-length protein such as the amino acid sequence shown in any one of the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539. Antibodies comprising amino acid residues and raised against a peptide include that epitope to form a specific immune complex with the full-length protein or any fragment containing the epitope. Preferably, the antigenic peptide comprises at least 10 amino acid residues, or at least 15 amino acid residues, or at least 20 amino acid residues, or at least 30 amino acid residues. Preferred epitopes encompassed by the antigenic peptides are protein regions located on the surface; these are usually hydrophilic regions.

  In certain embodiments of the invention, at least one epitope encompassed by the antigenic peptide is a NOVX region, eg, a hydrophilic region, localized on the surface of the protein. Hydrophobic analysis of the human NOVX protein sequence indicates which regions of the NOVX polypeptide are particularly hydrophilic and therefore likely to encode useful surface residues for targeting antibody production. As a means to target antibody production, hydropathy plots showing hydrophilic and hydrophobic regions are well known in the art, including, for example, the Kyte Doolittle or Hopp Woods methods, both with or without Fourier transforms. It can be generated by any method. For example, Hopp and Woods, 1981, Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte and Doolittle 1982, J. Mol. Biol. 157: 105-142 (both are hereby incorporated by reference in their entirety) As a part of the book). Antibodies that are specific for one or more domains within an antigenic protein or derivative, fragment, analog or homologue thereof are also provided herein.

  The proteins of the invention, or derivatives, fragments, analogs, homologues, or orthologs thereof can be utilized as an immunogen in the production of antibodies that immunospecifically bind to these protein components.

  Various methods known in the art can be used for the production of polyclonal or monoclonal antibodies against a protein of the invention, or derivatives, fragments, analogs, homologues, or orthologs thereof (eg, Antibodies: A Laboratory Manual). Harlow E, and Lane D, 1988, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, which is incorporated herein by reference). Some of these antibodies are discussed below.

Polyclonal antibodies For the production of polyclonal antibodies, various suitable host animals (eg rabbits, goats, mice, or other mammals) are injected with one or more natural proteins, synthetic variants thereof, or the aforementioned derivatives. To immunize. Suitable immunogenic formulations may include, for example, naturally occurring immunogenic proteins, chemically synthesized polypeptides that are representative of immunogenic proteins, or recombinantly expressed immunogenic proteins. Furthermore, the protein may be complexed with a second protein that is known to be immunogenic in the mammal to be immunized. Examples of such immunogenic proteins include, but are not limited to, keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. The formulation may further contain an adjuvant. Various adjuvants used to increase the immune response include Freund's (complete and incomplete) adjuvants, mineral gels (e.g. aluminum hydroxide), surfactants (e.g. lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions) , Dinitrophenol, etc.), adjuvants that can be used in humans, such as, but not limited to, Bacillus Calmette-Guerin and Corynebacterium parvum. Yet another example of an adjuvant that may be used may include MPL-TDM adjuvant (monophosphoryl lipid A, synthetic dicorynomycolic acid trehalose).

  Polyclonal antibodies against immunogenic proteins are isolated from mammals (eg, from blood) and known techniques, such as affinity chromatography using protein A or protein G, which provides mainly the IgG fraction in immune serum Can be used for further purification. Subsequently, or alternatively, a specific antigen that is the target of the desired immunoglobulin, or an epitope thereof, can be immobilized on a column and the immunospecific antibody can be purified by immunoaffinity chromatography. The purification of immunoglobulins is discussed, for example, by D. Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia PA, Vol. 14, No. 8 (April 17, 2000), pp. 25-28).

Monoclonal antibody As used herein, the term “monoclonal antibody” (MAb) or “monoclonal antibody composition” refers to only one molecular species of an antibody molecule comprising a characteristic light chain gene product and a characteristic heavy chain gene product. Refers to the population of antibody molecules it contains. In particular, the complementarity determining regions (CDRs) of a monoclonal antibody are identical in all molecules of the population. Thus, MAbs contain an antigen binding site capable of immunoreacting with a particular epitope of an antigen characterized by a characteristic binding activity thereto.

  Monoclonal antibodies can be prepared using hybridoma methods, as described by Kohler and Milstein, Nature, 256: 495 (1975). In the hybridoma method, lymphocytes that produce or are capable of producing antibodies that specifically bind to an immunizing agent are typically immunized by immunizing a mouse, hamster, or other suitable host animal with the immunizing agent. To trigger. Alternatively, lymphocytes can be immunized in vitro.

The immunizing agent typically includes protein antigens, fragments thereof or fusion proteins thereof. Generally, either peripheral blood lymphocytes are used if human-derived cells are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. . The lymphocytes are then fused with an immortal cell line using a suitable fusing agent such as polyethylene glycol to generate hybridoma cells (Goding, Monoclonal Antibodies: Principles and Practice , Academic Press, (1986) pp. 59 -103). Immortal cell lines are usually transformed mammalian cells, particularly myeloma cells from rodents, cows or humans. Usually, rat or mouse myeloma cell lines may be used. The hybridoma cells may be cultured in a suitable medium that preferably contains one or more substances that inhibit the growth or survival of unfused, immortalized cells. For example, if the parent cell lacks the enzyme hypoxanthine guanine phosphoribosyltransferase (HGPRT or HPRT), the culture medium for hybridomas typically contains hypoxanthine, a substance that prevents the growth of HGPRT-deficient cells, Contains aminopterin and thymidine (“HAT medium”).

  Preferred immortal cell lines are those that fuse efficiently, support high level expression of antibodies by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. Further preferred immortal cell lines are mouse myeloma cell lines available from, for example, the Salk Institute Cell Distribution Center, San Diego, California and the American Type Culture Collection, Manassas, Virginia. Human myeloma and mouse-human heteromyeloma cell lines are also described for the production of human monoclonal antibodies (Kozbor, J. Immunol., 133: 3001 (1984); Brodeur et al., Monoclonal Antibody Production Techniques and Applications, Marcel Dekker, Inc., New York, (1987) pp. 51-63).

  The medium in which the hybridoma is cultured can then be assayed for the presence of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by hybridoma cells is measured by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunosorbent assay (ELISA). Such techniques or assays are known in the art. The binding affinity of a monoclonal antibody can be measured, for example, by Scatchard analysis of Munson and Pollard, Anal. Biochem., 107: 220 (1980). Identifying antibodies with a high degree of specificity and high binding affinity for the target antigen is a particularly important objective in the therapeutic application of monoclonal antibodies.

  After identifying the desired hybridoma cells, the clones can be subcloned by limiting dilution and grown by standard methods (Goding, 1986). Suitable media for this purpose may include, for example, Dulbecco's Modified Eagle's Medium media and RPMI-1640 media. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

  Monoclonal antibodies secreted by subclones are isolated or isolated from medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein-A sepharose, hydroxyapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography. It can be purified.

  Monoclonal antibodies can also be made by recombinant DNA methods as described in US Pat. No. 4,816,567. Using an oligonucleotide probe capable of specifically binding DNA encoding the monoclonal antibody of the present invention to a gene encoding the heavy and light chains of the murine antibody using conventional procedures (eg. Can be easily isolated and sequenced. The hybridoma cells of the invention serve as a preferred source of such DNA. Once isolated, the DNA is placed in an expression vector which is then host cells such as monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that would otherwise not produce immunoglobulin proteins. It can be transfected into to obtain monoclonal antibody synthesis in recombinant host cells. DNA can also be replaced, for example, by replacing coding sequences for the normal domains of human heavy and light chains in place of homologous murine sequences (US Pat. No. 4,816,567; Morrison, Nature 368, 812- 13 (1994)), or all or part of the coding sequence for a non-immunoglobulin peptide can be modified by covalent attachment to the coding sequence of the immunoglobulin. Such non-immunoglobulin polypeptides can be substituted for the normal domain of the antibody of the invention, or can be substituted for the variable domain of one antigen binding site of the antibody of the invention to produce chimeric 2 A titer antibody can be created.

Humanized antibodies Antibodies to protein antigens of the present invention further include humanized antibodies or human antibodies. These antibodies are suitable for administration to humans without causing human immune responses to the administered immunoglobulins. Humanized forms of antibodies consist of chimeric immunoglobulins, immunoglobulin chains, or fragments thereof (Fv, Fab, Fab ′) that consist primarily of human immunoglobulin sequences and contain minimal sequence derived from non-human immunoglobulin. , F (ab ′) ₂ or other antigen-binding subsequence of the antibody). Humanization is performed by Winter or co-workers (Jones et al., Nature, 321: 522-525 (1986); Riechmann et al., Nature, 332: 323-327 (1988); Verhoeyen et al., Science, 239 : 1534-1536 (1988)) by replacing the rodent CDR or CDR sequence with the corresponding sequence of a human antibody. (See also US Pat. No. 5,225,539). In some cases, the Fv frame residues of the human immunoglobulin can be replaced by corresponding non-human residues. Humanized antibodies may also comprise residues that are not found in the recipient antibody or in the CDR or frame sequence. In general, a humanized antibody will contain at least one, and typically substantially all of the two variable domains. Here, all or substantially all of the CDR regions correspond to the CDR regions of a non-human immunoglobulin, and all or substantially all of the frame region is of a human immunoglobulin consensus sequence. A humanized antibody optimally also comprises at least a portion of an immunoglobulin normal region (Fc), typically that of a human immunoglobulin (Jones et al., 1986; Riechmann et al., 1988; and Presta , Curr. Op. Struct. Biol., 2: 593-596 (1992)).

Human antibodies Completely human antibodies relate essentially to antibody molecules in which the entire L and H chain sequences, including the CDRs, originate from a human gene. Such antibodies are referred to herein as “human antibodies” or “fully human antibodies”. Human monoclonal antibodies can be prepared by trioma technology; human B cell hybridoma technology (see Kozbor, et al., 1983 Immunol Today 4: 72) and EBV hybridoma technology to produce human monoclonal antibodies (Cole, et al. , 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). Humanized monoclonal antibodies can be utilized in the practice of the invention and human hybridomas can be used (see Cote, et al., 1983. Proc Natl Acad Sci USA 80: 2026-2030) or human B cells can be Can be produced by transformation with Epstein Barr virus (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96).

  In addition, human antibodies can also be obtained from phage display libraries (Hoogenboom and Winter, J. Mol. Biol., 227: 381 (1991); Marks et al., J. Mol. Biol., 222: 581 (1991)). Can be produced using further techniques including: Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, eg, mice. Here, the endogenous immunoglobulin genes are partially or completely inactivated. The challenge observes human antibody production very similar to that seen in humans in all aspects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in US Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016 And Marks et al. (Bio / Technology 10, 779-783 (1992)), Lonberg et al. (Nature 368 856-859 (1994)), Morrison (Nature 368, 812-13 (1994)), Fishwild et al, (Nature Biotechnology 14, 845-51 (1996)), Neuberger (Nature Biotechnology 14, 826 (1996)), Lonberg and Huszar (Intern. Rev. Immunol. 13 65-93 (1995)). .

  Human antibodies can additionally be produced using transgenic non-human animals that have been modified to produce fully human antibodies rather than the animal's endogenous antibodies in response to challenge with antigen (PCT Publication No. WO94 / 02602). The endogenous genes encoding immunoglobulin heavy and light chains in the non-human host are disabled, and active loci encoding human immunoglobulin heavy and light chains are inserted into the host genome. Human genes can be incorporated, for example, using yeast artificial chromosomes containing the necessary human DNA segments. Animals that provide all the desired modifications are then obtained as offspring by crossbreeding intermediate transgenic animals that contain less than the full complement of modifications. A preferred embodiment of such a non-human animal is a mouse, designated Xenomouse® as disclosed in PCT Publication Nos. WO96 / 33735 and WO96 / 34096. This animal produces B cells that secrete fully human immunoglobulins. The antibody can be obtained directly from the animal after immunization with the immunogen of interest, eg, as a polyclonal antibody specimen, or alternatively, an immortalized B derived from an animal, such as a hybridoma producing a monoclonal antibody. It can also be obtained from cells. In addition, genes encoding immunoglobulins with human variable regions can be recovered and expressed to obtain antibodies directly, or further modified, eg, for antibodies such as single chain Fv molecules. Analogues can be obtained.

  An example of a method of producing a non-human host, exemplified as a mouse, that lacks endogenous immunoglobulin heavy chain expression is disclosed in US Pat. No. 5,939,598. It is known from at least one endogenous heavy chain locus in embryonic stem cells to prevent loci rearrangement and to prevent transcript production of rearranged immunoglobulin heavy chain loci. A segment is deleted, this deletion being brought about by a targeting vector containing a gene encoding a selectable marker and somatic cells or embryonic cells are transformed from embryonic stem cells into the gene containing the gene encoding the selectable marker. It is obtained by a method comprising producing.

  Methods for producing antibodies of interest, such as human antibodies, are disclosed in US Pat. No. 5,916,771. It introduces an expression vector containing a nucleotide sequence encoding an H chain into one cultured mammalian host cell, and introduces an expression vector containing a nucleotide sequence encoding an L chain into yet another mammalian host cell. And fusing two cells to produce a hybrid cell. Hybrid cells express antibodies that contain heavy and light chains.

  In a further refinement of this procedure, a method for identifying clinically relevant epitopes on an immunogen and a correlative method for selecting antibodies that immunospecifically bind to a relevant epitope with high affinity is a PCT publication. No. WO99 / 53049.

_Fab fragments and single chain antibodies In accordance with the present invention, techniques can be adapted to the production of single chain antibodies specific for the antigenic proteins of the present invention (see, eg, US Pat. No. 4,946,778). In addition, the method is adapted to the construction of _Fab expression libraries (see, eg, Huse, et al., 1989 Science 246: 1275-1281), and to proteins or derivatives, fragments, analogs or homologues thereof. It may allow rapid and effective identification of monoclonal _Fab fragments with the desired specificity. Antibody fragments containing idiotypes against protein antigens can be produced by techniques known in the art, including (i) F _{(ab ′) 2} fragments produced by pepsin digestion of antibody molecules; (ii) F _{( ab ′) Fab} fragments obtained by reducing disulfide bridges of ₂ fragments; (iii) _Fab fragments generated by treating antibody molecules with papain and a reducing agent; and (iv) _Fv fragments, It is not limited to.

Bispecific antibodies Bispecific antibodies are monoclonal antibodies, preferably human or humanized antibodies, that have binding specificities for at least two different antigens. As used herein, one of the binding specificities is for the antigenic protein of the present invention. The second binding target is any other antigen, preferably a cell surface protein or receptor or receptor subunit.

  Methods for making bispecific antibodies are known in the art. Traditionally, recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy / light chain pairs in which the two heavy chains have different specificities (Milstein and Cuello, Nature, 305 : 537-539 (1983)). Due to the random combination of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture of ten different antibody molecules, only one of which has the correct bispecific structure. Purification of the correct molecule is usually performed by affinity chromatography. Similar methods are disclosed in WO 93/08829 published in May 1993 and Traunecker et al., EMBO J., 10: 3655-3659 (1991).

  Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin normal domain sequences. The fusion is preferably with the normal domain of an immunoglobulin heavy chain comprising at least part of the hinge, CH2 and CH3 regions. It is preferred to have a first heavy chain normal region (CH1) containing at least the site necessary for light chain binding present in one of the fusions. The DNA encoding the immunoglobulin heavy chain fusion and, if desired, the immunoglobulin light chain are inserted into separate expression vectors and transfected into a suitable host organism. For further details of bispecific antibodies see, for example, Suresh et al., Methods in Enzymology, 121: 210 (1986).

  According to another approach described in WO 96/27011, the interface between a pair of antibody molecules can be modified to maximize the percentage of heterodimer recovered from the recombinant cell medium. A preferred interface includes at least a portion of the CH3 region of the normal domain of an antibody. In this method, one or more small amino acid side chains at the interface of the first antibody molecule are replaced with larger side chains (eg tyrosine or tryptophan). The interface of the second antibody molecule by replacing a compensatory “void” of the same or similar size as the large side chain (s) by replacing the large amino acid side chain with a smaller one (eg alanine or threonine). To generate. This provides a mechanism to increase the yield of heterodimers over other undesirable end products such as homodimers.

Bispecific antibodies can be prepared as full length antibodies or antibody fragments (eg F (ab ′) ₂ bispecific antibodies). Techniques for generating bispecific antibodies from antibody fragments have been described in the literature. For example, bispecific antibody fragments can be prepared using chemical linkage. Brennan et al., Science 229: 81 (1985) describes a procedure in which intact antibodies are cleaved with proteolytic enzymes to produce F (ab ′) ₂ fragments. These fragments are reduced in the presence of the dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The resulting Fab ′ fragment is then converted to a thionitrobenzoic acid (TNB) derivative. One of the Fab′-TNBs is then reconverted to Fab′-thiol by reduction with mercaptoethylamine and mixed with an equal amount of other Fab′-TNB derivatives to produce bispecific antibodies. The resulting bispecific antibody can be used as an enzyme selective immobilization agent.

In addition, Fab ′ fragments can be recovered directly from E. coli and chemically coupled to produce bispecific antibodies. Shalaby et al., J. Exp. Med. 175: 217-225 (1992) describes the production of F (ab ′) ₂ molecules of fully humanized bispecific antibodies. Each Fab ′ fragment was secreted separately from E. coli and subjected to directed chemical coupling in vitro to form bispecific antibodies. Thus, the formed bispecific antibody can bind to cells that overexpress the ErbB2 receptor and normal human T cells and induce lytic activity of human cytotoxic lymphocytes against human breast tumor targets. did it.

Various techniques for making and isolating bispecific antibodies directly from recombinant cell media are also described. For example, bispecific antibodies are produced using leucine zippers. Kostelny et al., J. Immunol. 148 (5): 1547-1553 (1992). Leucine zippers from Fos protein and Jun protein were linked to the Fab ′ portions of two different antibodies by gene fusion. Antibody homodimers were reduced at the hinge region to form monomers and then reoxidized to form antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The “diabody” technology described by Hollinger et al., Proc. Natl. Acad. Sci. USA 90: 6444-6448 (1993) provides yet another mechanism for bispecific antibody fragment production. The fragment contains a heavy chain variable region (V _H ) linked to a light chain variable region (V _L ) by a linker, but does not allow pairing between two domains on the same chain due to the short linker . Thus, the V _H and V _L domains of one fragment are forced to pair with the complementary _VL and V _H domains of another fragment, thereby forming two antigen binding sites. Another strategy for making bispecific antibody fragments by the use of single chain Fv (sFv) dimers has also been reported. See Gruber et al., J. Immunol. 152: 5368 (1994).

An antibody having a valence of 3 or more can also be considered. For example, trispecific antibodies can be prepared. For example, Tutt et al., J. Immunol. 147: 60 (1991).
A typical bispecific antibody can bind to two different epitopes, at least one of which is derived from a protein antigen of the invention. Alternatively, the anti-antigen arm of an immunoglobulin molecule is linked to a T cell receptor molecule (e.g., CD2, CD3, CD28 or B7) or FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD16) on leukocytes. In combination with an arm that binds to an inducible molecule such as the Fc receptor for IgG (Fcγ), a cellular defense mechanism against cells expressing a particular antigen may be focused. Bispecific antibodies can also be used to direct cytotoxic agents to cells expressing a particular antigen. These antibodies possess an antigen binding arm and an arm that binds a cytotoxic agent or a radionuclide chelator such as EOTUBE, DPTA, DOTA or TETA. Another bispecific antibody of interest binds to the protein antigen described herein and further binds tissue factor (TF).

Heteroconjugate antibodies Heteroconjugate antibodies are also within the scope of the present invention. A heteroconjugate antibody consists of two antibodies linked by a covalent bond. Such antibodies are for example for targeting cells of the immune system to unfavorable cells (US Pat. No. 4,676,980) and for the treatment of HIV infection (WO91 / 00360; WO92 / 003733; EP03089). ). It is believed that these antibodies can be prepared in vitro using known methods of protein synthesis chemistry, including those involving crosslinkers. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose may include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in US Pat. No. 4,676,980.

Effector Functional Engineering For example, it is desirable to modify the antibodies of the present invention with respect to effector function in order to enhance the effectiveness of the antibody in the treatment of cancer. For example, cysteine residue (s) may be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated may have improved internalization ability and / or enhanced complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191-1195 (1992) and Shopes, J. Immunol., 148: 2918-2922 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linking agents as described in Wolff et al. Cancer Research, 53: 2560-2565 (1993). Alternatively, an antibody having a dual Fc region, thereby having enhanced complement-dependent cell lysis and ADCC ability, can be engineered. See Stevenson et al., Anti-Cancer Drug Design, 3: 219-230 (1989).

Immune complexes The present invention also includes chemotherapeutic agents, toxins (e.g., enzymatically active toxins of bacterial, fungal, plant, or animal origin, or fragments thereof) or radioisotopes (i.e., radioconjugates). It relates to an immunoconjugate comprising an antibody conjugated with such a cytotoxic substance.

Chemotherapeutic agents useful for the generation of such immune complexes are described above. Enzymatically active toxins and fragments thereof that can be used include diphtheria A chain, non-binding active fragments of diphtheria toxin, endotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, avirin A chain, modesin A chain, Alphasarcin, Aleurites fordii protein, dianthin protein, Phytolaca americana protein (PAPI, PAPII and PAP-S), momoridica charantia inhibitor, crucine, crotin, sapaonaria officinalisc inhibitor, gelonin, mitogerin, Listristine, phenomycin, enomycin and trichothecene. Various radionuclides are available for the production of radioconjugated antibodies. As examples, ²¹² Bi, ¹³¹ I, ¹³¹ In, ⁹⁰ Y and ¹⁸⁶ Re may be mentioned.

  A complex of an antibody and a cytotoxic agent is converted to a bifunctional derivative of N-succinimidyl-3- (2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), an imide ester (such as dimethyl adipimidate HCL Active esters (such as disuccinimidyl suberate), aldehydes (such as glutaraldehyde), bisazide derivatives (such as bis- (p-azidobenzoyl) -hexanediamine), bisdiazonium derivatives (such as bis -(Such as p-diazonium benzoyl) -ethylenediamine), diisocyanates (such as triene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro 2,4-dinitrobenzene) A variety of bifunctional protein coupling agents are used. For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science, 238: 1098 (1987). Carbon-14 labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is a typical chelating agent for conjugating radionucleotides to antibodies. See WO94 / 11026.

  In another embodiment, the antibody can be conjugated to a “receptor” (such as streptavidin) for pretargeting the tumor, wherein the antibody-receptor complex is administered to the patient. The unbound complex is then removed from the blood circulation using a scavenger and then a “ligand” (eg, avidin) conjugated to the cytotoxic agent is then administered.

Immunoliposome liposomes The antibodies disclosed herein can also be formulated as immunoliposome liposomes. Liposome liposomes containing antibodies are described in Epstein et al., Proc. Natl. Acad. Sci. USA, 82: 3688 (1985); Hwang et al., Proc. Natl Acad. Sci. USA, 77: 4030 (1980). And U.S. Pat. Nos. 4,485,045 and 4,544,545, as known in the art. Liposome liposomes with increased circulation time are disclosed in US Pat. No. 5,013,556.

  Particularly useful liposomal liposomes can be generated by the reverse phase evaporation method with a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposome liposomes are extruded through a filter with a defined pore size to obtain liposome liposomes of the desired diameter. Fab ′ fragments of the antibodies of the present invention can be bound to liposome liposomes via a disulfide exchange reaction as described in Martin et al., J. Biol. Chem., 257: 286-288 (1982). A chemotherapeutic agent (such as doxorubicin) is optionally contained within the liposome. See Gabizon et al., J. National Cancer Inst., 81 (19): 1484 (1989).

Diagnostic applications of antibodies directed against the proteins of the invention Antibodies directed against the proteins of the invention can be used for the localization and / or quantification of proteins (e.g. use in the measurement of protein levels in an appropriate physiological sample, It can be used in a manner known in the art regarding use in diagnostic methods, use in protein imaging, and the like. In certain embodiments, an antibody containing an antigen binding domain for a protein or derivative, fragment, analog, or homologue thereof is utilized as a pharmacologically active compound (see below).

Antibodies specific for the protein of the invention can be used to isolate the protein by standard techniques such as immunoaffinity chromatography or immunoprecipitation. Such antibodies can facilitate the purification of naturally derived proteins from cells and recombinantly produced antigens expressed in host cells. Furthermore, such antibodies can be used to detect antigenic proteins (eg, in cell lysates or cell supernatants) to assess the abundance or pattern of the antigenic protein. Antibodies directed against proteins can be used diagnostically to monitor protein levels in tissues as part of a clinical laboratory procedure, for example, to determine the effectiveness of a given treatment. Detection can be facilitated by coupling (ie, physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, combination groups, fluorescent materials, luminescent materials, bioluminescent materials and radioactive materials. Examples of suitable enzymes may include horseradish peroxidase, alkaline phosphatase, β-galactosidase and acetylcholinesterase; examples of suitable formulations may include streptavidin / biotin and avidin / biotin; Examples of umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; examples of luminescent materials include luminol Examples of bioluminescent materials may include luciferase, luciferin and aequorin, and examples of suitable radioactive materials may include ¹²⁵ I, ¹³¹ I, ³⁵ S or ³ H.

Antibody Therapies Antibodies of the invention, including polyclonal, monoclonal, humanized and fully human antibodies, can be used as therapeutic agents. Such agents are generally used for the treatment or prevention of the disease or lesion of interest. An antibody formulation, preferably one having high specificity and high affinity for its target antigen, is administered to a subject and generally has an effect due to binding to the target. Such an effect is one of two types depending on the specific nature of the interaction between a given antibody molecule and the target antigen in question. In the first case, administration of the antibody may prevent or inhibit binding of the target to the endogenous ligand to which it is bound. In this case, the antibody binds to the target and masks the naturally occurring ligand binding site where the ligand acts as an effector molecule. Thus, the receptor mediates the signaling pathway in which the ligand plays a role.

  Alternatively, the effect may be that the antibody elicits a physiological result by binding to an effector binding site on the target molecule. In this case, a target that is a receptor having an endogenous ligand that may not be present or defective in the disease or pathology may be combined with an antibody as an alternative effector ligand to produce a receptor-based signaling event. Initiated by the receptor.

  A therapeutically effective amount of an antibody of the invention generally relates to the amount required to achieve a therapeutic purpose. As mentioned above, this may be a binding interaction between an antibody and its target antigen that in certain cases inhibits the function of the target and in other cases promotes a physiological response. The amount required for administration will further depend on the binding affinity of the antibody for the particular antigen and also on the rate at which the administered antibody can be removed from the free volume of the administered subject. The usual range of therapeutically effective doses of an antibody or antibody fragment of the invention can be, by way of non-limiting example, from about 0.1 mg / kg body weight to about 50 mg / kg body weight. The usual number of administrations can range, for example, from twice a day to once a week.

Antibody Pharmaceutical Compositions Antibodies that specifically bind to the proteins of the invention, as well as other molecules identified by the screening assays disclosed herein, are administered in the form of pharmaceutical compositions for the treatment of various disorders. obtain. The principles and considerations involved in the preparation of such compositions, as well as guidelines for component selection, are described, for example, in The Science And Practice Of Pharmacy 19th ed. (Alfonso R. Gennaro, et al., Editors) Mack Pub. Co. ., Easton, Pa .: 1995, Drug Absorption Enhancement: Concepts, Possibilities, Limitations, And Trends, Harwood Academic Publishers, Langhorne, Pa., 1994, and Peptide And Protein Drug Delivery (Advances In Parenteral Sciences, Vol. 4), Provided in 1991, M. Dekker, New York.

  If the antigenic protein is intracellular and an intact antibody is used as an inhibitor, an internalizing antibody is preferred. However, liposomes can be used to deliver antibodies or antibody fragments into cells. When using antibody fragments, the smallest inhibitory fragment that specifically binds to the binding domain of the target protein is preferred. For example, peptide molecules can be designed that retain the ability to bind to the sequence of the target protein based on the sequence of the variable region of the antibody. Such peptides can be synthesized by chemical and / or recombinant DNA techniques. See, for example, Marasco et al., Proc. Natl. Acad. Sci. USA, 90: 7889-7893 (1993). The formulation herein may also contain more than one active compound as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. Alternatively or in addition, the composition may contain an agent that enhances its function, eg, a cytotoxic agent, cytokine, chemotherapeutic agent, or growth inhibitory agent. Such molecules are conveniently present in combination in amounts that are effective for the intended purpose.

  The active ingredients are colloidal drug delivery systems, for example in microcapsules prepared by coacervation technology or interfacial polymerization, for example in hydroxymethylcellulose or gelatin-microcapsules and poly- (methyl methacrylate) microcapsules, respectively. (E.g., liposomes, albumin spherules, microemulsions, nanoparticles and nanocapsules) or in macroemulsions.

The formulation to be used for in vivo administration must be sterile. This is easily accomplished by filtration through a sterile filtration membrane.
Sustained release formulations can be prepared. Suitable examples of sustained release formulations include solid-phase hydrophobic polymer semipermeable matrices containing antibodies, which are in the form of shaped articles such as films or microcapsules, for example. Yes. Examples of sustained release matrices include polyesters, hydrogels (eg, poly (2-hydroxyethyl-methacrylate) or poly (vinyl alcohol), polylactide (US Pat. No. 3,773,919), L-glutamic acid and Degradable lactic acid such as γ-ethyl-L-glutamate copolymer, non-degradable ethylene-vinyl acetate, LUPRON DEPOT® (injectable microspheres comprising lactic acid-glycolic acid copolymer and leuprolide acetate) Glycolic acid copolymers include polymers such as ethylene vinyl acetate and lactic acid-glycolic acid, which can release molecules over 100 days, while certain hydrogels release proteins in a shorter period of time.

ELISA Assay The agent that detects the analyte protein is an antibody capable of binding to the analyte protein, preferably an antibody with a detectable label. The antibody may be polyclonal or more preferably monoclonal. An intact antibody or a fragment thereof (eg, F _ab or F _{(ab) 2} ) can be used. The term “label” refers to a direct labeling of a probe or antibody by coupling (ie, physically linking) a detectable substance to the probe or antibody with respect to the probe or antibody, as well as another It is intended to include indirect labeling of probes or antibodies by reactivity with drugs. Examples of indirect labeling include detection of the first antibody using a fluorescently labeled second antibody and end labeling of the DNA probe with biotin so that it can be detected with fluorescently labeled streptavidin. The term “biological sample” is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. Thus, the term “biological sample” can include blood fractions or components including blood and serum, plasma, or lymph. That is, the detection methods of the present invention can be used to detect analyte mRNA, protein or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for the detection of analyte mRNA may include Northern hybridization and in situ hybridization. Analytical protein in vitro detection techniques may include enzyme-linked immunosorbent assay (ELISA), Western blot, immunoprecipitation and immunofluorescence. A technique for in vitro detection of analyte DNA may include Southern hybridization. The procedure for performing an immunoassay is described, for example, in `` ELISA: Theory and Practice: Methods in Molecular Biology '', Vol. 42, JR Crowther (Ed.) Human Press, Totowa, NJ, 1995, `` Immunoassay '', E. Diamandis and T. Christopoulus, Academic Press, Inc., San Diego, CA, 1996, and “Practice and Thory of Enzyme Immunoassays”, P. Tijssen, Elsevier Science Publishers, Amsterdam, 1985. In addition, techniques for in vivo detection of analyte proteins include introducing labeled anti-analyte protein antibodies into a subject. For example, the antibody can be labeled with a radioactive marker whose presence or localization in a subject can be detected by standard imaging techniques.

NOVX Recombinant Expression Vectors and Host Cells Another aspect of the present invention relates to vectors, preferably expression vectors, containing nucleic acids encoding NOVX proteins or derivatives, fragments, analogs or homologues thereof. As used herein, the term “vector” refers to a nucleic acid molecule capable of transporting another nucleic acid bound thereto. One type of vector is a “plasmid”, which refers to a circular double stranded DNA loop with additional DNA segments attached to it. Another type of vector is a viral vector, where additional DNA segments can be ligated into the viral genome. Certain vectors can be autonomously amplified in the host cell into which they are introduced (eg, bacterial vectors having a bacterial origin of replication and mammalian episomal vectors). Other vectors (eg, mammalian non-episomal vectors) can be integrated into the genome of a host cell upon introduction into the host cell, thereby replicating along with the host genome. In addition, certain vectors are capable of directing the expression of genes that are operably linked to them. Such vectors are referred to herein as “expression vectors”. In general, expression vectors useful in recombinant DNA technology are often in the form of plasmids. In the present specification, “plasmid” and “vector” may be used interchangeably as the plasmid is the most commonly used form of vector. However, the present invention is intended to include other forms of expression vectors such as viral vectors with equivalent functions (eg, retroviruses, adenoviruses and adeno-associated viruses lacking replication ability).

  A recombinant expression vector of the invention comprises a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which expresses a recombinant expression virus selected based on the host cell used for expression. It is meant to include one or more regulatory sequences operably linked to the nucleic acid sequence of interest. Within a recombinant expression vector, “operably linked” means that the nucleic acid sequence of interest is capable of expressing the nucleic acid sequence (eg, in an in vitro transcription / translation system or by introducing the vector into a host cell). In this case, it is meant to be bound to a regulatory sequence (in the host cell).

  The term “regulatory sequence” is intended to include promoters, enhancers and other expression control elements (eg, polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those that direct constitutive expression of nucleotide sequences in many types of host cells and those that direct expression of nucleotide sequences only in specific host cells (e.g., tissue-specific regulatory sequences). obtain. Those skilled in the art will appreciate that the design of the expression vector may depend on factors such as the choice of host cell to transform, the expression level of the desired protein, and the like. A protein or peptide comprising a fusion protein or peptide encoded by a nucleic acid as described herein (e.g., NOVX protein, mutant form of NOVX protein, fusion) by introducing an expression vector of the invention into a host cell Protein and the like).

  The recombinant expression vectors of the invention can be designed for NOVX protein expression in prokaryotic or eukaryotic cells. For example, NOVX protein can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors), yeast cells or mammalian cells. Suitable host cells are further discussed in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

  Protein expression in eukaryotic cells is most often performed using vectors containing constitutive or inducible promoters that direct the expression of either fusion or non-fusion proteins in Escherichia coli. Fusion vectors add a number of amino acids to the protein encoded therein, usually at the amino terminus of the recombinant protein. Such fusion vectors typically serve three purposes: (i) to increase expression of the recombinant protein; (ii) to increase the solubility of the recombinant protein; and (iii) in affinity chromatography. To help purify recombinant protein by acting as a ligand. Often, in fusion expression vectors, proteolytic cleavage sites are introduced at the junction of the fusion moiety and the recombinant protein to allow purification of the fusion protein following separation of the recombinant protein from the fusion moiety. Such enzymes, and their homologous recognition sequences, may include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988. Gene 67: 31) in which glutathione S-transferase (GST), maltose E-binding protein, or protein A is fused to a target recombinant protein, respectively. -40), pMAL (New England Biolabs, Beverly, Mass.) And pRIT5 (Pharmacia, Piscataway, NJ).

  Examples of suitable inducible non-fused E. coli expression vectors include pTrc (Amrann et al., (1988) Gene 69: 301-315) and pET11d (Studier et al., GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 60-89).

  One strategy to maximize protein expression in E. coli is to express the protein in a host cell that has impaired the ability to proteolytically cleave the recombinant protein. See Gottesman, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is to alter the nucleic acid sequence of the nucleic acid inserted into the expression vector so that individual codons for each amino acid are preferentially used in E. coli (eg, Wada, et al. al., 1992. Nucl. Acids Res. 20: 2111-2118). Such alteration of the nucleic acid sequences of the present invention can be made by standard DNA synthesis techniques.

  In another embodiment, the NOVX expression vector is a yeast expression vector. Examples of expression vectors in yeast Saccharomyces cerivisae include pYepSec1 (Baldari, et al., 1987. EMBO J. 6: 229-234), pMFa (Kurjan and Herskowitz, 1982. Cell 30: 933-943), pJRY88 (Schultz et al., 1987. Gene 54: 113-123), pYES2 (Invitrogen Corporation, San Diego, Calif.), And picZ (InVitrogen Corp, San Diego, Calif.).

  Alternatively, NOVX can be expressed in insect cells using baculovirus expression vectors. Baculovirus vectors that can be used for protein expression in cultured insect cells (eg, SF9 cells) include the pAc series (Smith, et al., 1983. Mol. Cell. Biol. 3: 2156-2165) and the pVL series. (Lucklow and Summers, 1989. Virology 170: 31-39).

  In yet another embodiment, the nucleic acids of the invention can be expressed in mammalian cells using mammalian expression vectors. Examples of mammalian expression vectors may include pCDM8 (Seed, 1987. Nature 329: 840) and pMT2PC (Kaufman, et al., 1987. EMBO J. 6: 187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2, cytomegalovirus and simian virus 40. For other expression systems suitable for both prokaryotic and eukaryotic cells, see, for example, Chapters 16 and 17 of Sambrook, et al., MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor See Laboratory Press, Cold Spring Harbor, NY, 1989.

  In another embodiment, the recombinant mammalian expression vector is capable of preferentially directing the expression of a nucleic acid in a particular cell type (e.g., using tissue specific regulatory elements to express the nucleic acid). ). Tissue specific regulatory elements are known in the art. Non-limiting examples of suitable tissue specific promoters include albumin promoters (liver specific; Pinkert, et al., 1987. Genes Dev. 1: 268-277), lymphoid specific promoters (Calame and Eaton, 1988. Adv. Immunol. 43: 235-275), especially T cell receptors (Winoto and Baltimore, 1989. EMBO J. 8: 729-733) and immunoglobulins (Banerji, et al., 1983. Cell 33: 729) -740; Queen and Baltimore, 1983. Cell 33: 741-748), neuron-specific promoters (eg, neurofilament promoters; Byrne and Ruddle, 1989. Proc. Natl. Acad. Sci. USA 86: 5473-5477 ), Pancreas-specific promoters (Edlund, et al., 1985. Science 230: 912-916), and mammary gland specific promoters (eg, whey; US Pat. No. 4,873,316 and European Patent Application No. 264, 166). For example, the developmentally regulated promoters of the mouse hox promoter (Kessel and Gruss, 1990. Science 249: 374-379) and the α-fetoprotein promoter (Campes and Tilghman, 1989. Genes Dev. 3: 537-546) Include.

  The present invention further provides a recombinant expression vector comprising a DNA molecule of the present invention cloned in an antisense orientation into the expression vector. That is, the DNA molecule is operably linked to regulatory sequences in a manner that allows expression of an RNA molecule that is antisense to NOVX mRNA (by transcription of the DNA molecule). Selecting regulatory sequences, such as viral promoters and / or enhancers, operably linked to the nucleic acid cloned into the antisense orientation that directs the sequential expression of the antisense RNA molecule in various cell types A regulatory sequence can be selected that directs tissue-specific or cell-type-specific constitutive expression of the antisense RNA. Antisense expression vectors can be in the form of recombinant plasmids, phagemids or attenuated viruses that produce antisense nucleic acids under the control of a highly efficient regulatory region whose activity is determined by the cell type into which the vector has been introduced. See, for example, Weintraub, et al., “Antisense RNA as a molecular tool for genetic analysis” Reviews-Trends in Genetics, Vol. 1 (1) 1986, for a discussion of the regulation of gene expression using antisense genes. I want.

  Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms “host cell” and “recombinant host cell” are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell, but also to the progeny or potential progeny of such a cell. Such progeny may not actually be identical to the parent cell, as the mutations or environmental effects may cause certain modifications in later generations, but the scope of the terms used herein Still included.

  The host cell can be any prokaryotic or eukaryotic cell. For example, NOVX protein can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are well known to those skilled in the art.

  Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms “transformation” and “transfection” refer to foreign nucleic acids (eg, DNA) including calcium phosphate or calcium chloride coprecipitation, DEAE-dextran mediated transfection, lipofection, or electroporation. It shall refer to various techniques recognized in the art for introduction into a host cell. Suitable methods for transforming or transfecting host cells include MOLECULAR CLONING: A LABORATORY MANUAL.2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989) and others. Can be found in the laboratory manual.

  For stable transfection of mammalian cells, depending on the expression vector used and the transfection technique, a small fraction of the cell can incorporate foreign DNA into its genome. In order to identify and select these integrants, a selectable marker (eg, resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Various selectable markers include those that confer resistance to drugs such as G418, hygromycin and methotrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding NOVX or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by drug selection (eg, cells that incorporate a selectable marker survive while other cells die).

  A host cell of the invention, such as a prokaryotic or eukaryotic cell in culture, can be used to produce (ie, express) NOVX protein. Therefore, the present invention further provides a method for producing NOVX protein using the host cell of the present invention. In one embodiment, the method comprises culturing a host cell of the invention (into which a recombinant expression vector encoding NOVX protein is introduced) in a suitable medium that produces NOVX protein. Including. In another embodiment, the method further comprises isolating NOVX protein from the medium or the host cell.

Transgenic NOVX Animals The host cells of the invention can also be used to produce non-human transgenic animals. For example, in one embodiment, the host cell of the invention is a fertilized oocyte or embryonic stem cell into which a NOVX protein coding sequence is introduced. A non-human transgenic animal that uses such a host cell to introduce an exogenous NOVX sequence into the genome, or a homologous recombinant animal that alters an endogenous NOVX sequence therein Can be created. Such animals are useful for studying NOVX protein function and / or activity and for identifying and / or evaluating modulators of NOVX protein activity. As used herein, a “transgenic animal” is a non-human animal, preferably a mammal, more preferably a rat or mouse, in which one or more cells of the animal contain the transgene. It is a tooth. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, and the like. The transgene is incorporated into the genome of the cell from which the transgenic animal develops and remains in the genome of the mature animal, thereby coding in one or more cell types or tissues of the transgenic animal. This is an exogenous DNA that directs the expression of a modified gene product. As used herein, “homologous recombinant animal” refers to an endogenous NOVX gene introduced into the endogenous gene and animal cells, eg, animal embryo cells, prior to animal development. A non-human animal, preferably a mammal, more preferably a mouse, which has been altered by homologous recombination with an exogenous DNA molecule.

  The transgenic animal of the present invention introduces a nucleic acid encoding NOVX into the male pronucleus of a fertilized oocyte (eg, by microinjection, retroviral infection), and the oocyte is injected into a pseudopregnant female foster animal. It can be created by allowing it to occur in the body. Any one human NOVX cDNA sequence of the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 can be introduced as a transgene into the genome of a non-human animal. Alternatively, a non-human homologue of the human NOVX gene, such as the mouse NOVX gene, can be isolated based on hybridization with human NOVX cDNA (detailed further above) and used as a transgene. Intron sequences and polyadenylation signals can also be included in the transgene to increase the expression efficiency of the transgene. Tissue specific regulatory sequences can be operably linked to the NOVX transgene to direct expression of the NOVX protein to specific cells. Methods for producing transgenic animals, particularly animals such as mice, by embryo manipulation and microinjection have become conventional in the art, for example, US Pat. No. 4,736,866; 4,870,009. And 4,873,191; and Hogan, 1986. In: MANIPULATING THE MOUSE EMBRYO, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. Similar methods can be used to produce other transgenic animals. Primary transgenic animals can be identified based on the presence of NOVX transgenes in the genome and / or expression of NOVX mRNA in animal tissues or cells. The primary transgenic animals can then be used to breed additional animals carrying the transgene. In addition, a transgenic animal carrying a transgene encoding a NOVX protein can be used to breed additional transgenic animals carrying other transgenes.

  A vector containing at least part of a NOVX gene into which deletions, additions or substitutions have been introduced in order to create homologous recombinant animals, thereby altering the NOVX gene, for example functionally disrupting To prepare. The NOVX gene can be a human gene (eg, any one cDNA of the group consisting of SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and SEQ ID NO: 538), more preferably human It is a non-human homologue of the NOVX gene. For example, using the mouse homologue of any one human NOVX gene in the group consisting of SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and SEQ ID NO: 538, the endogenous A homologous recombination vector suitable for altering the sex NOVX gene can be constructed. In one embodiment, the vector is designed to functionally disrupt an endogenous gene (ie, no longer encodes a functional protein; also referred to as a “knockout” vector) by homologous recombination.

  Alternatively, the vector mutates or otherwise alters the endogenous NOVX gene by homologous recombination, but still encodes a functional protein (e.g., alters the upstream regulatory region thereby changing the endogenous To alter the expression of sex NOVX protein). In a homologous recombination vector, the altered protein of the NOVX gene is bound to the endogenous NOVX gene and embryonic stem cells carried by the vector adjacent to the 5 'end or 3' end by additional nucleic acids of the NOVX gene. Allows homologous recombination to occur between NOVX genes. Yet another flanking NOVX nucleic acid is of sufficient length to allow successful homologous recombination with the endogenous gene. Typically, a few kilobases of contiguous DNA (both 5 'and 3' ends) can be included in the vector. For a description of homologous recombination vectors, see, for example, Thomas, et al., 1987. Cell 51: 503. The vector is then introduced into an embryonic stem cell line (eg, by electroporation) and cells in which the introduced NOVX gene is homologously recombined with the endogenous NOVX gene are selected. See, for example, Li, et al., 1992. Cell 69: 915.

  The selected cells are then injected into an animal (eg, mouse) blastocyst to form an aggregated chimera. See, for example, Bradley, 1987. In: TERATOCARCINOMAS AND EMBRYONIC STEM CELLS: A PRACTICAL APPROACH, Robertson, ed. IRL, Oxford, pp. 113-152. The chimeric embryo is then implanted into a suitable pseudopregnant female foster animal, and the embryo is delivered at term. Using progeny holding DNA homologously recombined in germ cells, all cells of the animal will breed animals containing homologous recombinant DNA by germline transmission of the transgene be able to. Methods for constructing homologous recombination vectors and homologous recombination animals are described in Bradley, 1991. Curr. Opin. Biotechnol. 2: 823-829; PCT International Publication No .: WO90 / 11354; WO91 / 01140; WO92 / 0968. And further described in WO 93/04169.

  In another embodiment, non-human transgenic animals can be produced that contain selected systems that allow for regulated expression of the transgene. One example of such a system is the cre / loxP recombinase system of bacteriophage P1. See, for example, Lakso, et al., 1992. Proc. Natl. Acad. Sci. USA 89: 6232-6236 for a description of the cre / loxP recombinase system. Another example of a recombinase system is the Saccharomyces cerevisiae FLP recombinase system. See O'Gorman, et al., 1991. Science 251: 1351-1355. If the cre / loxP recombinase system is used to regulate transgene expression, an animal containing a transgene encoding both the Cre recombinase and the selected protein is required. Such animals are, for example, “double” transgenic animals by mating two transgenic animals, one containing a transgene encoding the selected protein and the other containing a transgene encoding a recombinase. It can be provided by constructing.

Non-human transgenic animal clones described herein can also be produced according to the method described in Wilmut, et al., 1997. Nature 385: 810-813. Briefly, isolated cells from transgenic animals (e.g., somatic cells), induction, it is possible to enter the G ₀ phase exits from the growth cycle. The quiescent cells can then be fused with enucleated oocytes from the same type of animal from which the quiescent cells were isolated, for example, by use of electric pulses. The reconstituted oocyte is then cultured such that it develops into a morula or blastocyst and then introduced into a pseudopregnant female foster animal. The offspring born from this female foster animal is an animal clone that isolates cells (eg, somatic cells).

Pharmaceutical Compositions of pharmaceutical compositions suitable for administration of the NOVX nucleic acid molecules, NOVX proteins, anti-NOVX antibodies (also referred to herein as “active compounds”), and derivatives, fragments, analogs and homologs thereof of the present invention. Can be built in. Typically, such compositions comprise a nucleic acid molecule, protein or antibody and a pharmaceutically acceptable carrier. As used herein, “pharmaceutically acceptable carrier” refers to any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are compatible with pharmaceutical administration. Intended to include. Suitable carriers are described in the latest edition of Remington's pharmacology, a standard reference text in this field, which is hereby incorporated by reference. Preferred examples of such carriers or excipients include, but are not limited to, water, saline, finger solutions, glucose solutions and 5% human serum albumin. Water-insoluble vehicles such as liposomes and fatty oils are also used. Such media and agents for pharmaceutically active substances are well known in the art. Unless any conventional media or agents are compatible with the active compounds, their use in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

  A pharmaceutical composition of the invention is formulated to be compatible with its intended route of administration. Examples of routes of administration may include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g. inhalation), transdermal (i.e. topical), transmucosal and rectal administration. . Solutions or suspensions used for parenteral, intradermal or subcutaneous applications may include the following components: sterile excipients such as water for injections, saline solutions, fatty oils, polyethylene glycols, Glycerin, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl paraben; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); Buffers such as acid salts or phosphates, and agents that adjust isotonicity such as sodium chloride or glucose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple use vials made of glass or plastic.

  Pharmaceutical compositions suitable for injection may include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for preparations prepared as necessary in sterile injectable solutions or dispersions. For intravenous administration, suitable carriers may include physiological saline, bacteriostatic water, Cremophor EL® (BASF, Parsippany, N.J.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. Protection of microbial action is achieved by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, sodium chloride in the composition. Prolonged absorption of injectable compositions can be brought about by including in the composition an agent that delays absorption, for example, aluminum monostearate and gelatin.

  By incorporating the active compound (eg, a NOVX protein or anti-NOVX antibody) in the required amount, together with one or a combination of the above-listed ingredients, in a suitable solvent, and then sterile sterilizing, followed by filter sterilization Can be prepared. Generally, dispersions can be prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the method of preparation involves vacuum drying and freezing to produce a powder of the active ingredient plus any further desired ingredient powder from the pre-sterile filtered solution. It is dry.

  Oral compositions generally include an inert excipient or edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with additives and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a liquid carrier for use as a mouthwash, where the compound in the liquid carrier is applied orally, quickly removed, and exhaled or swallowed. Pharmaceutically compatible binding agents, and / or adjuvant materials can be included as part of the composition. Tablets, pills, capsules, troches and the like may contain any of the following ingredients or compounds with similar properties: binders such as microcrystalline cellulose, gum tragacanth or gelatin; additives such as starch or lactose, alginic acid Disintegrants such as primogel or corn starch; lubricants such as magnesium stearate or sterote; slip agents such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or mint, methyl salicylate, orange A fragrance like perfume.

  For administration by inhalation, the compounds can be delivered in the form of an aerosol spray from a pressurized container or dispenser which contains a suitable propellant, eg, a gas such as carbon dioxide, or a nebulizer.

  Systemic administration can also be obtained by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art and may include detergents, bile salts, fusidic acid derivatives, for example, for transmucosal administration. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, ointments, gels, or creams as generally known in the art.

  The compounds can also be prepared in the form of suppositories (eg, with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

  In one embodiment, the active compounds are prepared with carriers that will prevent the compound from rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable and biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polyacetic acid. It will be clear to those skilled in the art how to prepare such formulations. Materials are also commercially available from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted from infected cells with monoclonal antibodies to viral antibodies) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in US Pat. No. 4,522,811.

  It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. As used herein, unit dosage form refers to a physically discrete unit suitable as a single dosage for the subject being treated; each unit together with the required pharmaceutical carrier. Contains a predetermined amount of active compound calculated to produce the desired therapeutic effect. The unit dosage form specifications of the present invention are dictated by the unique features of the active compound and the particular therapeutic effect to be achieved, as well as limitations inherent in the art to formulate such active compounds for the treatment of individuals. And depends directly.

The nucleic acid molecule of the present invention can be inserted into a vector and used as a gene therapy vector. Gene therapy vectors can be obtained, for example, by intravenous injection, by local administration (see, eg, US Pat. No. 5,328,470) or by tactile injection (see, for example, Chen, et al., 1994. Proc. Natl. Acad. Sci. USA 91: 3054-3057). A pharmaceutical formulation of a gene therapy vector can include a gene therapy vector in an acceptable excipient, or can include a sustained matrix having the gene therapy vector embedded therein. Alternatively, where a complete gene therapy vector can be produced intact from a recombinant cell, such as a retrovirus vector, the pharmaceutical formulation can include one or more cells producing a gene delivery system.
The pharmaceutical formulation can be included in a container, pack or dispenser along with instructions for administration.

Screening and Detection Methods As detailed below, the isolated nucleic acid molecules of the invention are used to express NOVX protein (eg, via a recombinant expression vector in a host cell in gene therapy applications), Genetic defects in NOVX mRNA (eg, in a biological sample) or NOVX gene can be detected and NOVX activity can be modulated. In addition, NOVX protein is used to screen for drugs or compounds that modulate NOVX protein activity or expression, as well as poor or excessive production of NOVX protein, or reduced or abnormal compared to NOVX wild-type protein Diseases characterized by the production of NOVX protein types with various activities (eg; diabetes (modulating insulin release); obesity (lipid binding and transport); obesity-related metabolic disorders, metabolic syndrome X and chronic disorders And anorexia and debilitating disorders associated with various cancers, as well as infectious diseases (with antimicrobial activity) and various dyslipidemias.In addition, the anti-NOVX antibodies of the present invention may be used. NOVX protein can be detected and isolated, and NOVX activity can be modulated. Can be used in methods to affect appetite, nutrient absorption and disposal of metabolic substrates in both positive and negative ways.
The invention further relates to novel agents identified in the screening assays described herein and their use for the treatments described above.

Screening assays The present invention relates to modulators, i.e. candidate or test compounds or agents (e.g. peptides, peptides) that bind to NOVX protein or have a promoting or inhibitory effect on, for example, NOVX protein expression or NOVX protein activity. Methods for identifying mimetics, small molecules or other drugs (also referred to herein as “screening assays”) are provided. The invention also includes compounds identified in the screening assays described herein.

  In one embodiment, the invention provides an assay for screening candidate or test compounds that bind to or modulate the activity of a membrane-bound NOVX protein or polypeptide or biologically active portion thereof. To do. Test compounds of the present invention can be obtained from biological libraries; spatially addressable parallel solid or liquid phase libraries; synthetic library methods that require deconvolution; “one bead one compound” library Can be obtained using any of a number of approaches in combinatorial libraries well known in the art, including: methods; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds. See, for example, Lam, 1997. Anticancer Drug Design 12: 145.

  As used herein, “small molecule” refers to a composition having a molecular weight of less than about 5 kD, most preferably less than about 4 kD. Small molecules can be, for example, nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, lipids or other organic or inorganic molecules. Biological mixtures, such as chemical and / or fungal, bacterial, or algae extracts are known in the art and can be screened using any assay of the present invention.

  Examples of molecular library synthesis methods are described in, for example, DeWitt, et al., 1993. Proc. Natl. Acad. Sci. USA 90: 6909, Erb, et al., 1994. Proc. Natl. Acad. Sci. : 11422, Zuckermann, et al., 1994. J. Med. Chem. 37: 2678, Cho, et al., 1993. Science 261: 1303; Carrell, et al., 1994. Angew. Chem. Int. Ed. Engl. 33: 2059, Carell, et al., 1994. Angew. Chem. Int. Ed. Engl. 33: 2061, and Gallop, et al., 1994. J. Med. Chem. 37: 1233 Get a headline.

  A library of compounds can be obtained in solution (eg, Houghten, 1992. Biotechniques 13: 412-421) or on beads (Lam, 1991. Nature 354: 82-84) on a chip (Fodor, 1993. Nature 364: 555-556), bacteria (Ladner, US Pat. No. 5,223,409), spores (Ladner, US Pat. No. 5,233,409), plasmids (Cull, et al., 1992. Proc. Natl Acad. Sci. USA 89: 1865-1869) or on phage (Scott and Smith, 1990. Science 249: 386-390; Devlin, 1990. Science 249: 404-406, Cwirla, et al., 1990 Proc. Natl. Acad. Sci. USA 87: 6378-6382, Felici, 1991. J. Mol. Biol. 222: 301-310, Ladner, US Pat. No. 5,233,409).

In some embodiments, the assay is a cell-based assay, wherein a cell expressing a membrane-bound NOVX protein or biologically active portion thereof on the cell surface is contacted with the test compound and the test compound is present The ability to bind to NOVX protein is measured. The cell can be, for example, a mammal or a yeast cell. Measuring the ability of a test compound to bind to the NOVX protein can be accomplished, for example, by coupling the test compound with a radioisotope or enzyme label to bind the test compound to the NOVX protein or biologically active portion thereof. The measurement is made possible by detecting the labeled compound in the complex. For example, the test compound is labeled directly or indirectly with ¹²⁵ I, ³⁵ S, ¹⁴ C or ³ H, and the radioisotope is detected by direct counting of radiation emission or scintillation counting. Alternatively, the test compound is enzymatically labeled, for example, with horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzyme label is detected by measuring conversion of the appropriate substrate to product. In one embodiment, the assay comprises contacting a cell expressing a membrane-bound NOVX protein or biologically active portion thereof on the cell surface with a known compound that binds to NOVX to form an assay mixture. Contacting the assay mixture with the test compound and measuring the ability of the test compound to interact with the NOVX protein, wherein measuring the ability of the test compound to interact with the NOVX protein is determined by the test Measuring the ability of a compound to bind preferentially to a NOVX protein or biologically active portion thereof compared to a known compound.

  In another embodiment, the assay is a cell-based assay, contacting a cell that expresses a membrane-bound NOVX protein or biologically active portion thereof on the cell surface with a test compound, and the test compound is NOVX Measuring the ability to modulate (eg, promote or inhibit) the activity of a protein or biologically active portion thereof. Measuring the activity of a test compound that modulates the activity of NOVX or a biologically active portion thereof is, for example, by measuring the ability of a NOVX protein to bind to or interact with a NOVX target molecule. Can be achieved. As used herein, a “target molecule” is a molecule that binds or interacts with a natural NOVX or NOVX protein, such as a cell surface molecule that expresses a protein that interacts with a NOVX or NOVX, A cell surface molecule, a molecule in the extracellular environment, a molecule associated with the inner surface of the cell membrane or a cytoplasmic molecule. The target molecule for NOVX can be a non-NOVX molecule or a NOVX that is a NOVX protein or polypeptide of the invention. In one embodiment, a NOVX target molecule is a signal transduction that facilitates the transmission of extracellular signals (eg, signals generated by the binding of compounds to membrane-bound NOVX molecules) through and into the cell membrane. It is a component of the pathway. The target can be, for example, a second intracellular protein with catalytic activity or a protein that promotes the association of a downstream signal molecule with NOVX.

Measuring the ability of a NOVX protein to bind to or interact with a NOVX target molecule can be accomplished by one of the methods described above that measure direct binding. In one embodiment, measuring the ability of a NOVX protein to bind or interact with a target molecule of NOVX or NOVX can be accomplished by measuring the activity of the target molecule. For example, detecting the activity of a target molecule, detecting the induction of a target cellular second messenger (ie, intracellular Ca ²⁺ , diacylglycerol, IP ₃ etc.), detecting the catalytic / enzymatic activity of the target against the appropriate substrate Detecting the induction of a receptor gene (including a NOVX-responsive regulatory element operably linked to a nucleic acid encoding a detectable marker, such as luciferase), or a cellular response, such as a cell Can be measured by detecting cell survival, cell differentiation, or cell proliferation.

  In yet another embodiment, the assay of the present invention is a cell-free assay, wherein a NOVX protein or biologically active portion thereof is contacted with a test compound, and the test compound is NOVX protein or biological thereof Measuring the ability to bind to an active moiety. Binding of the test compound to the NOVX protein can be measured either directly or indirectly as described above. In one such embodiment, the assay comprises contacting the NOVX protein or biologically active portion thereof with a known compound that binds NOVX to form an assay mixture, contacting the assay mixture with the test compound. And measuring the ability of a test compound to interact with a NOVX protein, wherein measuring the ability of a test compound to interact with a NOVX protein includes determining that the test compound is a NOVX protein or organism thereof. Measuring the ability to bind preferentially to a chemically active moiety compared to a known compound.

  In yet another embodiment, the assay contacts a NOVX protein or biologically active protein thereof with a test compound, and the test compound modulates the activity of the NOVX protein or biologically active protein thereof ( A cell-free assay that involves measuring the ability to (eg promote or inhibit). Measuring the ability of a test compound to modulate the activity of NOVX can be achieved, for example, by one of the methods described above for measuring direct binding to the NOVX protein binding activity to a target molecule of NOVX. it can. In yet another embodiment, measuring the ability of a test compound to modulate the activity of a NOVX protein can be accomplished by measuring the ability of the NOVX protein to modulate an additional NOVX target molecule. For example, the catalytic / enzymatic activity of the target molecule against the appropriate substrate can be measured as described above.

  In yet another embodiment, the cell-free assay comprises contacting the NOVX protein or biologically active portion thereof with a known compound that binds to the NOVX protein to form an assay mixture, wherein the assay mixture is a test compound. And measuring the ability of the test compound to interact with the NOVX protein, wherein measuring the ability of the test compound to interact with the NOVX protein is the target of the NOVX with the NOVX protein. Measuring the ability to bind preferentially to a molecule or modulate its activity.

The cell-free assay of the present invention can be used for both soluble or membrane-bound NOVX proteins. In the case of cell-free assays involving membrane-bound NOVX protein, it is desirable to utilize a solubilizing agent so that the membrane-bound NOVX protein is maintained in solution. Examples of such solubilizers include n-octyl glucoside, n-dodecyl glucoside, n-dodecyl maltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton® X- 100, Triton® X-114, Thesit®, non-ionic surfactants such as isotridecipoly (ethylene glycol ether) _n , N-dodecyl-N, N-dimethyl-ammonio-1-propanesulfonate , 3- (3-Chloramidopropyl) dimethylaminol-1-propanesulfonate (CHAPS), or 3- (3-Chloramidopropyl) dimethylaminiol-2hydroxy-1-propanesulfonate (CHAPSO).

In two or more embodiments of the above assay methods of the invention, the NOVX protein or target molecule thereof may be immobilized to facilitate separation of the complex form from the uncomplexed form of one or both proteins, as well as the assay. Can adapt to automation. Binding of the test compound to the NOVX protein or the interaction of the NOVX protein with the target molecule in the presence and absence of the candidate compound can be achieved in any container suitable for containing the reactants. Examples of such containers may include microtiter plates, test tubes, and microcentrifuge tubes. In one embodiment, a fusion protein can be provided that adds a domain that allows one or both proteins to be bound to a matrix. For example, GST-NOVX fusion protein or GST-target fusion protein is adsorbed onto a microtiter plate derivatized with glutathione Sepharose beads (Sigma Chemical, St. Louis, MO) or glutathione, which is then tested or tested The compound and either non-adsorbed target protein or NOVX protein are bound and the mixture is incubated under conditions that promote complex formation (eg, physiological conditions with respect to salt and pH). After incubation, the beads or microtiter plate holes are washed to remove any unbound components, in the case of beads, the matrix is immobilized, and the complex is measured directly or indirectly, eg as described above. . Alternatively, the complex can be dissociated from the matrix and the level of NOVX protein binding or activity measured using standard techniques.

  Other techniques for immobilizing proteins on a matrix can also be used in the screening assays of the invention. For example, either the NOVX protein or its target molecule can be immobilized utilizing the binding of biotin and streptavidin. A biotinylated NOVX protein or target molecule is prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well known in the art (eg, biotinylated kit, Pierce Chemicals, Rockford, Ill.) To prepare streptavidin Can be immobilized in holes in a 96-well plate coated with (Pierce Chemical). Alternatively, an antibody that is reactive with the NOVX protein or target molecule but does not interfere with the binding of the NOVX protein to the target protein is derivatized into a hole in the plate to bind the unbound target or NOVX protein to the antibody. Can be captured in the hole. Methods for detecting such complexes include, in addition to those described above for GST-immobilized complexes, immunodetection of complexes using antibodies that react with NOVX proteins or target molecules, and NOVX proteins or target molecules. Mention may be made of enzyme binding assays that rely on detecting the relevant enzyme activity.

  In another embodiment, a modulator of NOVX protein expression is identified by a method of contacting a cell with a candidate compound and measuring the expression of NOVX mRNA or protein in the cell. The expression level of NOVX mRNA or protein in the presence of the candidate compound is compared to the expression level of NOVX mRNA or protein in the absence of the candidate compound. Candidate compounds can then be identified as NOVX mRNA or protein modulators based on this comparison. For example, a candidate compound is identified as a promoter of NOVX mRNA or protein expression when the expression of NOVX mRNA or protein is greater in the presence of the candidate compound than in the absence (ie, statistically significantly higher) . Alternatively, if the expression of NOVX mRNA or protein is less in the presence of the candidate compound (ie, statistically significantly less) in the presence of the candidate compound, the candidate compound may be an inhibitor of NOVX mRNA or protein expression. Identify as The level of expression of NOVX mRNA or protein in the cell can be measured by the methods described herein for detecting NOVX mRNA or protein.

  In yet another embodiment of the invention, the NOVX protein is a two-hybrid assay or a three-hybrid assay (eg, US Pat. No. 5,283,317, Zervos, et al., 1993. Cell 72: 223-232, Madura, et al., 1993). J. Biol. Chem. 268: 12046-12054, Bartel, et al., 1993. Biotechniques 14: 920-924, Iwabuchi, et al., 1993. Oncogene 8: 1693-1696, and Brent WO 94/10300. Can be used to identify other proteins that bind or interact with NOVX to modulate NOVX activity (“NOVX binding protein” or “NOVX-bp”). Such NOVX binding proteins are also involved, for example, in signal amplification by NOVX proteins as elements upstream or downstream of the NOVX pathway.

The two-hybrid system is based on the modular nature of most transcription factors consisting of separable DNA binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one construct, the gene encoding NOVX is fused to a gene encoding the DNA binding domain of a known transcription factor (eg, GAL-4). In other constructs, DNA from a DNA sequence library encoding an unidentified protein ("prey" or "sample") is fused to a gene encoding the activation domain of a known transcription factor. If the “bait” and “prey” proteins can interact in vivo to form a NOVX-dependent complex, the DNA binding and activation domains of transcription factors can be drawn closer together. This proximity allows transcription of a reporter gene (eg, LacZ) operably linked to a transcription factor responsive transcriptional regulatory site. Reporter gene expression can be detected, cell colonies containing functional transcription factors can be isolated and used to obtain cloned genes encoding proteins that interact with NOVX.
The invention further relates to novel agents identified by the aforementioned screening assays and their use in the treatments described herein.

Detection Assays A portion or fragment of the cDNA identified herein (and the corresponding complete gene sequence) can be used in various ways as a polynucleotide reagent. By way of example, and without limitation, these sequences: (i) map each gene on the chromosome; thus locate the gene region associated with the genetic disease; (ii) from a trace biological sample It can be used to help identify forensic identification of individuals (tissue typing); and (iii) biological samples. Some of these applications are described in the following subsections.

Chromosome mapping Once a sequence (or part of a sequence) of a gene is isolated, this sequence can be used to map the position of the gene on the chromosome. This method is called chromosome mapping. Thus, using a portion or fragment of a NOVX sequence selected from the group consisting of SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and SEQ ID NO: 538, or a fragment or derivative thereof, Each location of the NOVX gene on the chromosome can be mapped. Mapping NOVX sequences to the chromosome is an important first step in correlating these sequences with genes associated with disease.

  Briefly, the NOVX gene can be mapped to a chromosome by preparing PCR primers (preferably 15-25 bp long) from the NOVX sequence. Computer analysis of NOVX sequences can be used to quickly select primers that span no more than two exons in genomic DNA, thus complicating the amplification process. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Those hybrids containing the human gene corresponding to the NOVX sequence produce an amplified fragment.

  Somatic cell hybrids are prepared by fusing somatic cells from different mammals (eg, human and mouse cells). As hybrids of human and mouse cells grow and divide, they gradually delete human chromosomes in a random order, but retain mouse chromosomes. Mouse cells cannot grow because they lack specific enzymes, but human cells retain one human chromosome containing the gene encoding the required enzyme by using a medium that can grow. A panel of hybrid cell lines can be established by using various media. Each cell line in the panel contains a single human chromosome or a small number of human chromosomes, and a complete set of mouse chromosomes, allowing individual genes to be easily mapped to specific human chromosomes. See, for example, D'Eustachio, et al., 1983. Science 220: 919-924. Somatic cell hybrids containing only fragments of human chromosomes can also be produced by using human chromosomes with translocations and deletions.

  PCR mapping of somatic cell hybrids is a rapid procedure that maps a specific sequence to a specific chromosome. Using a single thermal cycler, it is possible to associate three or more sequences per day. By designing oligonucleotide primers with NOVX sequences, a detailed localization site specification can be achieved using a panel of fragments from specific chromosomes.

  Fluorescence in situ hybridization (FISH) of DNA sequences to metaphase chromosome spreads can further be used to provide precise chromosomal locations in one step. Chromosome spreads can be generated using cells that are blocked at metaphase by chemicals that disrupt the spindle, such as colcemid. Chromosomes can be briefly treated with trypsin and then Giemsa stained. A pattern of light and dark bands appears on each chromosome so that the chromosomes can be individually identified. FISH technology can be used with DNA sequences as short as 500 or 600 bases. However, clones larger than 1,000 bases are more likely to bind to specific chromosomal locations with signal intensity sufficient for simple detection. Preferably 1,000 bases and more preferably 2,000 bases are sufficient to obtain good results in a reasonable time. For a review of this technology, see Verma, et al., HUMAN CHROMOSOMES: A MANUAL OF BASIC TECHNIQUES (Pergamon Press, New York 1988).

  Use individual chromosome mapping reagents to label a single chromosome or a single site on that chromosome, or use a panel of reagents to label multiple sites and / or multiple chromosomes Can do. Reagents corresponding to non-coding regions of the gene are actually preferred for mapping purposes. The coding region is more likely to be conserved within the gene family, thus increasing the chance of cross-hybridization during chromosomal mapping.

  Once a sequence is mapped to a precise chromosomal location, the physical location of the sequence on the chromosome can be correlated with genetic map data. Such data is found, for example, in McKusick, Mendelian Inheritance in Man (available online through Hopkins University Welch Medical Library). The relationship between the genes mapped to the same chromosomal site and the disease can then be determined using the association analysis described in, for example, Egeland, et al., 1987. Nature, 325: 783-787 (for physically adjacent genes). Can be identified by co-inheritance).

  In addition, DNA sequence differences between individuals suffering from and not affected by diseases associated with the NOVX gene can be measured. If the mutation is observed in some or all of the affected individuals but not in any of the unaffected individuals, the mutation is likely a causative agent for a particular disease. Comparison of affected and unaffected individuals is generally first detected by using visible structural changes in the chromosome, such as deletions or translocations visible from the chromosome spread, or using PCR based on its DNA sequence With the search for possible structural changes. Finally, complete sequencing of genes from several individuals can be performed to confirm the presence of the mutation and to distinguish the mutation from the polymorphism.

Tissue typing The NOVX sequences of the invention can also be used to identify individuals from trace biological samples. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes and probed on a Southern blot to produce a unique band for identification. The sequences of the present invention are useful as yet another DNA marker for RFLP (restriction fragment length polymorphism, described in US Pat. No. 5,272,057).

  Furthermore, the sequences of the present invention may be used to provide yet another technique for measuring the actual base-by-base DNA sequence of a selected portion of an individual's genome. Thus, using the NOVX sequence described herein, two PCR primers can be prepared from the 5 'and 3' ends of the sequence. These primers can then be used to amplify the individual's DNA and subsequently sequence it.

  Since each individual has a characteristic set of such DNA due to allelic differences, a panel of corresponding DNA sequences from individuals prepared in this manner can provide identification of characteristic individuals. The sequences of the present invention can be used to obtain such identification sequences from individuals and tissues. The NOVX sequences of the present invention characteristically represent portions of the human genome. Allelic variation occurs to some extent in the coding regions of these sequences and to a greater extent in the non-coding regions. It is believed that allelic variation between individual humans occurs at a frequency of about 1 in 500 bases. Many of the allelic variations are due to single nucleotide polymorphisms (SNPs), including restriction fragment length polymorphisms (RFLP).

  Each of the sequences described herein can be used to some extent as a standard by which DNA from individuals can be compared for identification purposes. Since more genetic polymorphisms occur in non-coding regions, fewer sequences are required to identify individuals. Non-coding sequences can reasonably provide unambiguous identification of an individual with a panel of perhaps 10 to 1,000 primers, each resulting in a 100 base non-coding amplified sequence. If the predicted coding sequence is used, such as SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and any one sequence of the group consisting of SEQ ID NO: 538, an obvious individual A more suitable number of primers for identification is 500-2,000.

Predictive Medicine The present invention also relates to the field of predictive medicine, using diagnostic assays, prognostic assays, pharmacogenomics, and clinical trial monitoring for prognostic (predictive) purposes, thereby prophylactically treating an individual. Accordingly, one aspect of the present invention is to measure NOVX protein and / or nucleic acid expression and NOVX activity in the context of a biological sample (eg, blood, serum, cells, tissue), thereby causing an individual to become abnormal Relates to a diagnostic assay for determining whether or not one is suffering from or at risk of developing a disorder associated with the expression or activity of NOVX. Disorders include metabolic disorders, diabetes, obesity, infectious diseases, anorexia, cancer-related cachexia, cancer, neurodegenerative disorders, Alzheimer's disease, Parkinson's disease, immune disorders, and hematopoietic disorders, and various different fats May include metabolic disorders associated with blood pressure, obesity, metabolic syndrome X and debilitating diseases associated with chronic diseases and various cancers. The invention also provides a prognostic (predictive) assay to determine whether an individual is at risk for developing a disorder associated with NOVX protein, nucleic acid expression or activity. For example, certain NOVX gene mutations can be assayed in a biological sample. Such assays are used for prognostic or predictive purposes, thereby prophylacticizing individuals prior to the development of a disorder characterized or associated with NOVX protein, nucleic acid expression or biological activity. Can be treated.

  Another aspect of the present invention is a method for measuring NOVX protein, nucleic acid expression or activity in an individual, thereby selecting an appropriate therapeutic or prophylactic agent for that individual (referred to herein as “pharmacogenomics”). Provided). Pharmacogenomics allows the selection of an agent (e.g., a drug) for therapeutic or prophylactic treatment of an individual based on the individual's genotype (e.g., examining an individual's genotype to identify an individual's specific drug) Measure ability to respond to).

Yet another aspect of the invention relates to monitoring the effect of a drug (eg, drug, compound) on NOVX expression or activity in clinical trials.
These and other drugs are described in detail in the following sections.

Diagnostic Assays An exemplary method for detecting the presence or absence of NOVX in a biological sample is to obtain a biological sample from a test subject, and to use the biological sample as a NOVX protein or a nucleic acid encoding NOVX protein. Involves contacting a compound or agent capable of detecting (eg, mRNA, genomic DNA) to allow the presence of NOVX to be detected in a biological sample. An agent for detection of NOVX mRNA or genomic DNA is a labeled nucleic acid probe capable of hybridizing to NOVX mRNA or genomic DNA. The nucleic acid probe is, for example, a full-length NOVX nucleic acid such as SEQ ID NO: 2n-1 (where n is an integer from 1 to 129) and any one nucleic acid of the group consisting of SEQ ID NO: 538, or at least 15, 30 , 50, 100, 250 or 500 nucleotides in length and part of an oligonucleotide sufficient to specifically hybridize to NOVX mRNA or genomic DNA under stringent conditions. Other suitable probes for use in the diagnostic assays of the invention are described herein.

The agent for detecting NOVX protein is an antibody capable of binding to NOVX protein, preferably an antibody with a detectable label. The antibody may be polyclonal, or more preferably monoclonal. Intact antibodies, or fragments thereof (eg, Fab or F (ab ′) ₂ ) can be used. The term “label” for a probe or antibody refers to directly labeling a probe or antibody by coupling (ie, physically linking) a detectable substance to the probe or antibody, as well as another directly labeled Indirect labeling of the probe or antibody by reactivity with one reagent. Examples of indirect labeling may include detecting the first antibody using a fluorescently labeled second antibody and end-labeling the DNA probe with biotin so that it can be detected with fluorescently labeled streptavidin. . The term “biological sample” includes tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection methods of the invention can be used to detect NOVX mRNA, protein, or genomic DNA in a biological sample in vitro and in vivo. For example, in vitro techniques for detection of NOVX mRNA may include Northern hybridization and in situ hybridization. In vitro techniques for detecting NOVX protein may include enzyme linked immunoassay (ELISA), Western blot, immunoprecipitation, and immunofluorescence. In vitro techniques for detecting NOVX genomic DNA may include Southern hybridization. Further, in vivo techniques for detecting NOVX protein include introducing labeled anti-NOVX antibodies into the subject. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected with standard imaging techniques.

  In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a peripheral blood leukocyte sample isolated from a subject by conventional means.

  In another embodiment, the method comprises obtaining a control biological sample from a control subject, using the control sample as a compound or agent capable of detecting NOVX protein, mRNA or genomic DNA, and NOVX protein, mRNA or genome. Further contacting to detect the presence of DNA in the biological sample and comparing the presence of NOVX protein, mRNA or genomic DNA in the control sample to NOVX protein, mRNA or genomic DNA in the test sample Accompany.

  The invention also includes a kit for detecting the presence of NOVX in a biological sample. For example, the kit: a labeled compound or agent capable of detecting NOVX protein or mRNA in a biological sample; means for measuring the amount of NOVX in the sample; and means for comparing the amount of NOVX in the sample with a standard Can be included. The compound and drug can be packaged in a suitable container. The kit may further comprise instructions for use of the kit for detecting NOVX protein or nucleic acid.

Prognostic Assays Further, the diagnostic methods described herein can be used to identify subjects who have or are at risk of developing a disease or disorder associated with abnormal NOVX expression or activity. For example, using the assays described herein, such as previous diagnostic assays and the following assays, to identify subjects who have or are at risk of developing a disorder related to NOVX protein, nucleic acid expression or activity Can do. Alternatively, prognostic assays can be utilized to identify subjects having or at risk for developing a disease or disorder. Thus, the present invention provides a method for identifying a disease or disorder associated with aberrant NOVX expression or activity that obtains a test sample from a subject and detects aberrant NOVX protein or nucleic acid (eg, mRNA, genomic DNA). Wherein the presence of NOVX protein or nucleic acid is a diagnosis for a subject having or at risk of developing a disease or disorder associated with abnormal NOVX expression or activity. As used herein, “test sample” refers to a biological sample obtained from a subject of interest. For example, the test sample can be a biological fluid (eg, serum), a cell sample, or a tissue.

  In addition, the prognostic assays described herein can be used to treat agents (eg, agonists, antagonists, peptidomimetics, proteins, peptides) to treat diseases or disorders associated with abnormal NOVX expression or activity. , Nucleic acids, small molecules, or other pharmaceutical candidates) can be determined. For example, such methods can be used to determine whether a subject's disease is effectively treated with a drug. Thus, the present invention provides a method for obtaining a test sample and determining whether an agent can effectively treat a disorder associated with abnormal NOVX expression or activity that detects NOVX protein or nucleic acid. (For example, where the presence of NOVX protein or nucleic acid becomes a diagnosis for a subject who can be administered an agent to treat a disorder associated with abnormal NOVX expression or activity).

  The method of the invention may also be used to detect genetic damage in certain NOVX genes, such that the subject with the damaged gene is at risk for a disorder characterized by abnormal cell growth and / or differentiation Can decide. In various embodiments, these methods are genetically characterized in at least one alteration that affects the integrity of a gene encoding a NOVX protein, or misexpression of a NOVX gene, in a cell sample from a subject. Includes detection of the presence or absence of damage. For example, such genetic damage may include (i) deletion of one or more nucleotides from a NOVX gene; (ii) addition of one or more nucleotides to a NOVX gene; (iii) Substitution of one or more nucleotides of a NOVX gene, (iv) chromosomal rearrangement of a NOVX gene, (v) changes in mRNA transcript level of a NOVX gene, (vi) methylation pattern of genomic DNA, etc. (Vii) presence of a non-wild type splicing pattern of an mRNA transcript of a NOVX gene, (viii) a non-wild type level of a NOVX protein, (ix) an allelic deletion of a NOVX gene And (x) can be detected by confirmation of the presence of at least one inappropriate post-translational modification of a NOVX protein. There are a number of assay techniques known in the art that can be used to detect damage to certain NOVX genes described herein. A preferred biological sample is a peripheral blood leukocyte sample isolated from a subject by conventional means. However, any biological sample containing nucleated cells including, for example, buccal mucosa cells can be used.

  In certain embodiments, damage detection is in a polymerase chain reaction (PCR) such as anchor PCR or RACE PCR (see, eg, US Pat. Nos. 4,683,195 and 4,683,202) or Alternatively, Ligation Chain Reaction (LCR) (eg, Landegran, et al., 1988. Science 241: 1077-1080, and Nakazawa, et al., 1994. Proc. Natl. Acad. Sci. USA 91: 360 With the use of probes / primers in (see -364), but the latter may be particularly useful for the detection of point mutations in the NOVX gene (Abravaya, et al., 1995. Nucl. Acids Res. 23: 675-682). See). This method involves collecting a cell sample from a patient, isolating nucleic acid (eg, genome, mRNA or both) from the sample cells, and under conditions such that hybridization and amplification of the NOVX gene (if present) occurs. Contacting the nucleic acid sample with one or more primers that specifically hybridize to a NOVX gene, and detecting the presence or absence of the amplification product, or detecting the size of the amplification product, and length with the control sample A step of comparing the lengths. PCR and / or LCR are desirably used as a preliminary amplification step in conjunction with any technique used to detect mutations described herein.

  Further amplification methods include: self-retaining sequence replication (see Guatelli, et al., 1990. Proc. Natl. Acad. Sci. USA 87: 1874-1878), transcription amplification system (Kwoh, et al., 1989). Proc. Natl. Acad. Sci. USA 86: 1173-1177); Qβ replicase (see Lizardi, et al, 1988. BioTechnology 6: 1197), or any other nucleic acid amplification method followed by one skilled in the art May include detection of amplified molecules using well-known techniques. These detection schemes are particularly useful for the detection of such molecules if only a small number of nucleic acid molecules are present.

  In yet another embodiment, mutations in certain NOVX genes from sample cells can be identified by changes in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, amplified (optionally), treated with one or more restriction endonucleases, and fragment sizes are measured and compared by gel electrophoresis. Differences in fragment size between sample and control DNA indicate mutations in the sample DNA. In addition, sequence specific ribozymes (see, eg, US Pat. No. 5,493,531) can be used to assess the presence of specific mutations by the generation or disappearance of ribozyme cleavage sites.

  In other embodiments, genetic mutations in NOVX are identified by hybridizing sample and control nucleic acids, eg, DNA or RNA, to a high density array containing hundreds or thousands of oligonucleotide probes. can do. See, for example, Cronin, et al., 1996. Human Mutation 7: 244-255, Kozal, et al., 1996. Nat. Med. 2: 753-759. For example, genetic variations in NOVX can be identified in a two-dimensional array containing DNA probes that generate light as described in Cronin, et al., Supra. Briefly, the first hybridization array of probes is used to scan a long stretch of DNA in a sample and a control to create a series of overlapping probe linear arrays to determine base changes between sequences. Can be identified. This step allows the identification of point mutations. This is followed by a second hybridization that allows the characterization of a particular mutation by using a smaller special probe array that is complementary to all the mutants or mutations detected. Each mutation array consists of a set of parallel probes, one complementary to the wild type gene and the other complementary to the mutant gene.

  In yet another embodiment, the NOVX gene is directly sequenced using any of a variety of sequencing reactions known in the art, and the NOVX sequence of the sample is compared with the corresponding wild type (control) sequence. Mutations can be detected by comparison. Examples of sequencing reactions are based on the technique developed by Maxim and Gilbert, 1977. Proc. Natl. Acad. Sci. USA 74: 560 or Sanger, 1977. Proc. Natl. Acad. Sci. USA 74: 5463 You can list things. Sequencing by mass spectrometry (eg PCT International Publication No. WO 94/16101; Cohen, et al., 1996. Adv. Chromatography 36: 127-162 and Griffin, et al., 1993. Appl. Biochem. Biotechnol. 38: It can also be envisaged that any of a variety of automated sequencing methods, including 147-159) can be utilized in performing a diagnostic assay (see, eg, Naeve, et al., 1995. Biotechniques 19: 448).

Other methods for detecting mutations in the NOVX gene may include methods for detecting mismatched bases in RNA / RNA or RNA / DNA heteroduplexes using protection from cleaving agents. See, for example, Myers, et al., 1985. Science 230: 1242. In general, the art of “mismatch cleavage” is formed by hybridizing (labeled) RNA or DNA containing a wild-type NOVX sequence with RNA or DNA of a potential mutant obtained from a tissue sample. Starting with providing a heteroduplex. The duplex is treated with an agent that cleaves the single-stranded region of the duplex present due to base mismatches between the control and sample strands. For example, the RNA / DNA duplex is treated with RNase, DNA / DNA hybrids treated with _{S 1} nuclease may process the mismatched regions enzymatically. In other embodiments, either DNA / DNA or RNA / DNA duplexes can be treated with hydroxylamine or osmium tetroxide and piperidine to digest mismatched regions. After processing the mismatch region, the resulting material is then separated by size on a denaturing polyacrylamide gel to determine the site of mutation. See, for example, Cotton, et al., 1988. Proc. Natl. Acad. Sci. USA 85: 4397, Saleeba, et al., 1992. Methods Enzymol. 217: 286-295. In one embodiment, the control DNA or RNA can be labeled for detection.

  In yet another embodiment, the mismatch cleavage reaction recognizes mismatched base pairs in double stranded DNA in a defined system to detect and map point mutations in NOVX cDNA obtained from a sample of cells. One or more proteins (so-called “DNA mismatch repair” enzymes) are used. For example, E. coli mutY enzyme cleaves G / A mismatch A and thymidine DNA glycosidase from HeLa cells cleaves G / T mismatch T. See, for example, Hsu, et al., 1994. Carcinogenesis 15: 1657-1662. According to an exemplary embodiment, a probe based on a NOVX sequence, eg, a wild type NOVX sequence, is hybridized with cDNA or other DNA product from the test cell (s). This duplex is treated with a DNA mismatch repair enzyme, and if a cleavage product is present, it can be detected from an electrophoresis protocol or the like. See, for example, US Pat. No. 5,459,039.

  In other embodiments, changes in electrophoretic mobility are used to identify mutations in the NOVX gene. For example, single strand conformation polymorphism (SSCP) can be used to detect electrophoretic mobility differences between mutant and wild type nucleic acids. For example, Orita, et al., 1989. Proc. Natl. Acad. Sci. USA: 86: 2766, Cotton, 1993. Mutat. Res. 285: 125-144, Hayashi, 1992. Genet. Anal. Tech. Appl. 9: See 73-79. Allows single-stranded DNA fragments of sample and control NOVX nucleic acids to be denatured and renatured. The secondary structure of single-stranded nucleic acids varies from sequence to sequence, and the resulting change in electrophoretic mobility makes it possible to detect even single base changes. DNA fragments can be labeled or detected with a labeled probe. The sensitivity of the assay can be increased by using RNA (rather than DNA) whose secondary structure is more sensitive to sequence changes. In one embodiment, the subject method utilizes heteroduplex analysis to separate duplex helical duplex molecules based on changes in electrophoretic mobility. See, for example, Keen, et al., 1991. Trends Genet. 7: 5.

  In yet another embodiment, the migration of mutant or wild-type fragments in a polyacrylamide gel containing a gradient of denaturing agent is assayed using denaturing concentration gradient gel electrophoresis (DGGE). See, for example, Myers, et al., 1985. Nature 313: 495. When using DGGE as an analytical method, DNA is modified to ensure that it is not completely denatured, for example by adding a GC clamp of high melting point GC-enriched DNA of approximately 40 bp by PCR. In a further embodiment, temperature gradients are used instead of denaturing gradients to identify differences in the mobility of control and sample DNA. See, for example, Rosenbaum and Reissner, 1987. Biophys. Chem. 265: 12753.

  Examples of other techniques for detecting point mutations may include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension. For example, an oligonucleotide primer centered on a known mutation is prepared and then hybridized with the target DNA under conditions that allow hybridization only when a perfect match is found. See, for example, Saiki, et al., 1986. Nature 324: 163, Saiki, et al., 1989. Proc. Natl. Acad. Sci. USA 86: 6230. When attaching oligonucleotides to the hybridizing membrane and hybridizing to labeled target DNA, such allele-specific oligonucleotides hybridize to PCR amplified target DNA or a number of different mutants. .

  Alternatively, allele specific amplification techniques based on selective PCR amplification may be used in conjunction with the present invention. Oligonucleotides used as primers for specific amplification allow the mutation of interest to be centered in the molecule (as amplification depends on differential hybridization; see, eg, Gibbs, et al., 1989. Nucl. Acids Res. 17: 2437-2448) or, under appropriate conditions, mismatches can prevent or reduce polymerase extension (see, eg, Prossner, 1993. Tibtech. 11: 238) held at the 3 ′ extreme end of one primer. It may be. In addition, it is desirable to introduce new restriction sites in the region of the mutation in order to create detection based on cleavage. See, for example, Gasparini, et al., 1992. Mol. Cell Probes 6: 1. In certain embodiments, it is anticipated that amplification can also be performed using Taq ligase for amplification. See, for example, Barany, 1991. Proc. Natl. Acad. Sci. USA 88: 189. In such cases, ligation occurs only when there is a perfect match at the 3 'end of the 5' end sequence, and the presence of a known mutation at a particular site is detected by examining the presence or absence of amplification. Enable.

  The methods described herein can be performed, for example, by utilizing a prepacked diagnostic kit that includes at least one probe nucleic acid or antibody reagent described herein. This can be conveniently used, for example, in the clinical setting to diagnose a patient who exhibits symptoms or symptoms of a disease or disorder involving the NOVX gene or has a family history.

  In addition, any cell type or tissue that expresses NOVX, preferably peripheral blood leukocytes, may be utilized in the prognostic assays described herein. However, any biological sample containing, for example, nucleated cells of buccal mucosa cells may be used.

Pharmacogenomics A drug or modulator that has a stimulatory or inhibitory effect on NOVX activity (eg, NOVX gene expression) as identified by the screening assays described herein is administered to an individual to cause a disorder (including a metabolic disorder). , Diabetes, obesity, infectious disease, anorexia, cancer-related cachexia, cancer, neurodegenerative disorders, Alzheimer's disease, Parkinson's disease, immune disorders, and hematopoietic disorders, and various dyslipidemias, obesity Metabolic syndrome X associated with metabolic disorders, chronic diseases and various cancers as well as debilitating diseases can be treated (prophylactically or therapeutically). Along with such treatment, an individual's pharmacogenomics (ie, a study of the relationship between an individual's genotype and the individual's response to a foreign compound or drug) may be considered. Differences in therapeutic drug metabolism can result in severe toxicity or treatment failure by altering the relationship between pharmacologically active drug dose and blood concentration. Thus, the pharmacogenomics of an individual allows the selection of an effective agent (eg, drug) for prophylactic or therapeutic treatment based on consideration of the individual's genotype. Such pharmacogenomics can further be used to determine the appropriate dose and method of treatment. Accordingly, the activity (s) of NOVX protein, the expression of NOVX nucleic acids, or the content of NOVX gene mutations in an individual can be measured, thereby selecting the agent (s) appropriate for the therapeutic or prophylactic treatment of the individual.

  Pharmacogenomics deals with clinically significant genetic variation in response to drugs due to altered drug distribution and abnormal effects in affected individuals. See, for example, Eichelbaum, 1996. Clin. Exp. Pharmacol. Physiol., 23: 983-985; Linder, 1997. Clin. Chem., 43: 254-266. In general, two types of pharmacogenetic abnormalities can be distinguished. Genetic abnormalities that transmit as a single factor that changes the way the drug acts on the body (altered drug action) or genetic abnormalities that transmit as a single factor that changes the way the body acts on the drug (altered drug metabolism) There is. These pharmacogenetic abnormalities can occur as either rare defects or polymorphisms. For example, glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common hereditary enzyme disorder, in which the main clinical complications are oxidant drugs (antimalarials, sulfonamides) , Analgesics, nitrofurans) hemolysis after ingestion or after eating broad beans.

  In an exemplary embodiment, the activity of a drug's metabolic enzyme is a major determinant of both the intensity and duration of the drug's action. The discovery of genetic polymorphisms in drug metabolizing enzymes (eg, N-acetyltransferase 2 (NAT2) and cytochrome pregnancy band precursor protein enzymes CYP2D6 and CYP2C19) Provide an explanation of whether the expected effect of the drug is not achieved or if it shows excessive drug response and severe toxicity. These polymorphisms are expressed in the population population in two phenotypes: a high metabolic group (EM) and a low metabolic group (PM). The abundance of PM varies between different populations. For example, the gene encoding CYP2D6 is highly polymorphic and several mutations identify PMs that result in the absence of functional CYP2D6. The hypometabolic group of CYP2D6 and CYP2C19 very frequently experience excessive drug response and side effects when receiving standard doses. If the metabolite is an active therapeutic component, PM does not show a therapeutic response, as demonstrated by the analgesic action of codeine mediated by the metabolite morphine produced by CYP2D6. The exact opposite is the so-called ultrafast metabolic group that does not respond to standard doses. Recently, the molecular basis of the ultrafast metabolic group was confirmed to be due to CYP2D6 gene amplification.

  Thus, an individual's NOVX protein activity, NOVX nucleic acid expression, or mutation content of the NOVX gene can be measured, thereby selecting an appropriate agent (s) for therapeutic or prophylactic treatment of the individual . In addition, pharmacogenetic studies can be used to apply genotyping of polymorphic alleles encoding drug metabolizing enzymes to the identification of individual drug responsiveness phenotypes. In applying this knowledge to dosage and drug selection, adverse effects or treatment failures may be observed when treating a subject with a NOVX modulator, such as a modulator identified by one of the exemplary screening assays described herein. Avoiding and thus enhancing therapeutic or prophylactic efficiency.

Monitoring the effects of clinical trials Monitoring the impact of a drug (eg, drug, compound) on NOVX expression or activity (eg, activity that modulates abnormal cell growth and / or differentiation) is a fundamental drug screen. It can be applied not only to clinical trials. For example, screening assays as described herein can reduce the effectiveness of agents determined to increase NOVX gene expression, increase protein levels, or upregulate NOVX activity, to reduce NOVX gene expression, protein levels, or lower It can be monitored in clinical trials of subjects exhibiting controlled NOVX activity. Alternatively, the efficacy of an agent that has been determined to decrease NOVX gene expression, protein levels, or down-regulate NOVX activity is compared to the clinical efficacy of a subject exhibiting increased NOVX gene expression, protein levels, or upregulated NOVX activity. Can be monitored in the test. In such clinical trials, expression or activity of NOVX, and preferably other genes associated with, for example, cell proliferation or immune disorders, may be used as a “readout” or marker of immune responsiveness of specific cells. .

  By way of non-limiting example, a gene comprising NOVX that is modulated in a cell by treatment with an agent (eg, a compound, drug or small molecule) that modulates NOVX activity (eg, identified in a screening assay described herein) Can be identified. Thus, to examine the effects of drugs on cell proliferation disorders, for example, in clinical trials, cells can be isolated and RNA prepared and the expression levels of NOVX and other genes thought to be associated with the disorder can be analyzed. The amount of protein produced by gene expression levels (ie gene expression patterns) by Northern blot analysis or RT-PCR as described herein, or alternatively by one of the methods described herein. Or by measuring the activity level of NOVX or other genes. In this manner, gene expression patterns serve as markers that indicate the cellular physiological response to the drug. Thus, this response state can be measured before and during treatment of an individual with a drug at various times.

  In one embodiment, the invention is directed to drugs (e.g., agonists, antagonists, proteins, peptides, peptidomimetics, nucleic acids, small molecules, or other drug candidates identified in the screening assays described herein). A method is provided for monitoring the effectiveness of treating a subject, comprising: (i) obtaining a pre-dose sample from a subject prior to drug administration; (ii) NOVX protein, mRNA, or in a pre-dose sample; Detecting the expression level of genomic DNA, (iii) obtaining one or more post-administration samples from the subject, and (iv) the level of expression or activity of NOVX protein, mRNA, or genomic DNA in the post-administration sample (V) one or more samples after administration of the level of expression or activity of NOVX protein, mRNA, or genomic DNA in the sample prior to administration The comparison with NOVX protein, mRNA or genomic DNA,, and comprising the step of changing the administration of the agent to a subject according to (vi) it. For example, to increase NOVX expression or activity to a higher level than detected, ie, to increase the effectiveness of the drug, increased administration of the drug is desirable. Alternatively, in order to reduce NOVX expression or activity to a lower level than detected, ie, reduce the effectiveness of the drug, reduced administration of the drug is desirable.

Methods of Treatment The present invention provides both prophylactic and therapeutic methods for treating subjects at risk (susceptibility) or having a disease associated with abnormal NOVX expression or activity. Diseases include cardiomyopathy, atherosclerosis, hypertension, congenital heart disease, aortic stenosis, atrial septal defect (ASD), atrioventricular (AV) duct defect, arterial duct, pulmonary valve stenosis, Aortic lower stenosis, ventricular septal defect (VSD), valve disease, tuberous sclerosis, scleroderma, obesity, transplantation, adrenoleukodystrophy, congenital adrenal hyperplasia, prostate cancer, neoplasm, adenocarcinoma , Lymphoma, uterine cancer, fertility, hemophilia, hypercoagulability, idiopathic thrombocytopenic purpura, immunodeficiency, graft-versus-host disease, AIDS, bronchial asthma, Crohn's disease; multiple sclerosis, al Mention of the treatment of Bright hereditary bone dystrophy and other similar diseases, disorders and abnormalities may be mentioned.
These treatments are discussed in further detail below.

Diseases and Disorders Therapeutic agents that antagonize (i.e. reduce or inhibit) activity of diseases and disorders characterized by increased levels (as compared to subjects not suffering from the disease or disorder) or biological activity Can be treated with. A therapeutic that antagonizes activity may be administered in a therapeutic or prophylactic manner. The therapeutic agents that can be used include: (i) the aforementioned peptides, or analogs, derivatives, fragments or homologues thereof; (ii) antibodies to the aforementioned peptides; (iii) nucleic acids encoding the aforementioned peptides; (iv) Antisense nucleic acids and nucleic acids that are “dysfunctional” used to “knock out” the endogenous functions of the aforementioned peptides by homologous recombination (ie, heterologous into the coding sequence of the coding sequence for the aforementioned peptide (See, for example, Capecchi, 1989. Science 244: 1288-1292); or (v) a modulator that alters the interaction of the aforementioned peptide and its binding partner (ie, further of the invention Inhibitors, agonists, antagonists) including, but not limited to, mimetics of other peptides or antibodies specific for the peptides of the present invention.

  Diseases and disorders characterized by decreased levels (as compared to subjects not afflicted with the disease or disorder) or biological activity can be treated with therapeutic agents that increase activity (ie, are agonists). A therapeutic agent that upregulates activity may be administered in a therapeutic or prophylactic manner. The therapeutic agents that can be utilized can include, but are not limited to, the aforementioned peptides, or analogs, derivatives, fragments, or homologues thereof; or agonists that increase bioavailability.

  Increased or decreased levels are obtained from patient tissue samples (e.g., from biopsy tissue) and assayed in vitro for RNA or peptide levels, expressed peptide (or mRNA for the aforementioned peptides) structure and / or activity By doing so, it can be easily detected by quantifying peptides and / or RNA. Methods well known in the art include to detect immunoassay (eg, Western blot analysis, immunoprecipitation followed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, immunocytochemistry, etc.) and / or mRNA expression. Hybridization assays such as, but not limited to, Northern assays, dot blots, in situ hybridization, and the like.

Prophylactic methods In one aspect, by administering to a subject an agent that modulates abnormal NOVX expression or at least some NOVX activity, the present invention provides a method for treating a disease or disorder associated with abnormal NOVX expression or activity in a subject. Provide a way to prevent. Identifying a subject at risk for a disease caused or contributed to by aberrant NOVX expression or activity, eg, by any of diagnostic or prognostic assays or combinations thereof as described herein Can do. A prophylactic agent may be administered prior to the appearance of symptoms characteristic of NOVX abnormalities so as to prevent or alternatively delay the progression of the disease or disorder. Depending on the type of NOVX abnormality, for example, an agent that is a NOVX agonist or NOVX antagonist may be used to treat the subject. Appropriate agents can be determined based on the screening methods described herein. The prophylactic methods of the present invention are further discussed in the following subsections.

Therapeutic Methods Another aspect of the present invention relates to methods of modulating NOVX expression or activity for therapeutic purposes. The modulating method of the present invention comprises contacting a cell with an agent that modulates one or more activities of NOVX protein activity associated with the cell. An agent that modulates NOVX protein activity can be an agent described herein, such as a nucleic acid or protein, a naturally-occurring ligand of a NOVX protein, a peptide, an NOVX peptide mimetic, or a small molecule. In one embodiment, the agent promotes one or more NOVX protein activities. Examples of such facilitating agents may include active NOVX proteins and nucleic acid molecules encoding NOVX introduced into cells. In another embodiment, the agent inhibits one or more NOVX protein activities. Examples of such inhibitory agents may include antisense NOVX nucleic acid molecules and anti-NOVX antibodies. These adjustment methods can be performed in vitro (eg, by culturing cells with the agent) or alternatively, in vivo (eg, by administering the agent to the subject). Thus, the present invention provides a method of treating an individual suffering from a disease or disorder characterized by abnormal expression or activity of certain NOVX proteins or nucleic acid molecules. In one embodiment, the method comprises a combination of agents (e.g., agents identified in the screening assays described herein) or agents that modulate (e.g., upregulate or downregulate) NOVX expression or activity. With administration. In another embodiment, the method involves administering a NOVX protein or nucleic acid molecule as a treatment to compensate for decreased or abnormal NOVX expression or activity.

  In situations where NOVX is abnormally down-regulated and / or increased NOVX activity has a beneficial effect, it is desirable to promote NOVX activity. One example of such a situation is when the subject has a disorder characterized by abnormal cell proliferation and / or differentiation (eg, cancer or an immune related disease). Another example of such a situation is when the subject has a gestational disorder (eg, preeclampsia).

Measuring the biological effect of a therapeutic agent In various embodiments of the invention, appropriate in vitro or in vivo assays are performed to indicate the effect of a particular therapeutic agent and treatment of the tissue affected by its administration. Determine whether or not.

  In various specific embodiments, an in vitro assay is performed on a representative type (s) of cells involved in the patient's disorder to determine whether the administered therapeutic agent has the desired effect on the cell type. Can be measured. The compounds used for treatment can be tested in a suitable animal model system including, without limitation, rats, mice, chickens, cows, monkeys, rabbits, etc. prior to testing in human subjects. Similarly, for in vivo studies, any animal model system known in the art can be used prior to administration to a human subject.

Prophylactic and therapeutic uses of the compositions of the invention NOVX nucleic acids and proteins of the invention include: metabolic disorders, diabetes, obesity, infectious diseases, anorexia, cancer-related, neurodegenerative disorders, Alzheimer's disease, Parkinson's disease, Various disorders including but not limited to immune disorders, hematopoietic disorders, and various dyslipidemias, metabolic disorders associated with obesity, metabolic syndrome X and chronic diseases and various cancer-related debilitating diseases Useful for related potential prophylactic and therapeutic applications.

  By way of example, a cDNA encoding a NOVX protein of the invention can be useful for gene therapy, and the protein can be useful when administered to a subject in need thereof. By way of non-limiting example, the composition of the present invention may be used in metabolic diseases, diabetes, obesity, infectious diseases, anorexia, cancer-related cachexia, cancer, neurodegenerative disorders, Alzheimer's disease, Parkinson's disease, immunity It will have an effect on the treatment of patients suffering from disorders, hematopoietic disorders, and various dyslipidemias.

  Both novel nucleic acids encoding NOVX proteins and NOVX proteins of the invention, or fragments thereof, may also be useful in diagnostic applications to assess the presence or amount of nucleic acids or proteins. A further use may be as an antibacterial molecule (ie, certain peptides have been found to have antibacterial properties). These substances are further useful for the production of antibodies that immunospecifically bind to the novel substances of the invention for use in therapeutic or diagnostic methods.

Sequence analysis The sequence of NOVX is brought about by laboratory cloning of a cDNA fragment that is a computer prediction of the sequence. A cDNA fragment covering the full length of the DNA sequence, part of the sequence, or both was cloned. Predictions provide either full-length DNA sequences or portions thereof based on sequences available in the CuraGen exclusive sequence database or in public human sequence databases.
Experimental cloning was performed using one or more methods summarized below.

  SeqCalling® technology: cDNA was derived from a variety of human samples representing a variety of tissue types, normal and disease states, physiological states, and developmental states from different donors. Samples were obtained as whole tissue, early cells, or tissue cultured early cells or cell lines. Cells and cell lines were treated with biological or chemical reagents that modulate gene expression, eg, growth factors, chemokines, steroids. Thus, the cDNA is fully published in US Serial No. 09 / 417,386, filed October 13, 1999, and 09 / 614,505, filed July 11, 2000. Sequencing was performed using SeqCalling technology from CuraGen Corporation. Sequence traces were evaluated manually and the collection was edited if necessary. DNA sequences from all samples, sometimes including public human sequences, were collected using a bioinformatic program to produce a continuous sequence of each assembly. Each assembly contains a database of CuraGen Corporation. A sequence is included as a component of an assembly if the limit of identity with another component is at least 95%, greater than 50 bp. Each assembly represents those genes or parts and includes information on splice forms, single nucleotide polymorphisms (SNPs), variants such as insertions, deletions, and other sequence variants.

  Variant sequences are also included in the present invention. Variant sequences can include single nucleotide polymorphisms (SNPs). The SNP is sometimes referred to as “cSNP” to indicate that the nucleotide sequence containing the SNP starts as a cDNA. SNPs can occur in a variety of ways. For example, a SNP can contribute to the substitution of one nucleotide for another at a polymorphic site. Such substitutions can be either transitions or base substitutions. SNPs can also result from deletions or insertions of nucleotides associated with the reference allele. In this case, a polymorphic site is a site where one allele has a gap with respect to a particular nucleotide in another allele. SNPs that occur within a gene lead to changes in the amino acids encoded by the gene at the position of the SNP. Intragenic SNPs can also be silent if the codon containing the SNP encodes the same amino acid as a result of the redundancy of the genetic code. SNPs that occur outside the gene region or in the intron of the gene do not lead to changes in any amino acid sequence of the protein, but can lead to altered control of the expression pattern. Examples include changes in temporal expression, physiological response control, cell type expression control, intensity of expression, and stability of transcribed messengers.

  The information presented includes genomic clones, public genes and disclosed sequences and ESTs that share sequence identity with CuraGen Corporation's electronic northern bioinformatics tool.

Examples Example A: Polynucleotide and polypeptide sequences and homology data Example 1
The NOV1 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 1A.

Further analysis of the NOV1a protein resulted in the following properties shown in Table 1B.

A search for the NOV1a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 1C.

In the BLAST search of the public sequence database, the NOV1a protein was found to have homology with the proteins shown in the BLASTP data in Table 1D.

PFam analysis indicated that the NOV1a protein contains the domains shown in Table 1E.

Example 2
The NOV2 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 2A.

Continuation of Table 2A

Continuation of Table 2A

By comparing the protein sequences, the following sequence relationships shown in Table 2B were obtained.

Further analysis of the NOV2a protein resulted in the following properties shown in Table 2C.

A search for the NOV2a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 2D.

In a BLAST search of the public sequence database, the NOV2a protein was found to be homologous to the proteins shown in the BLASTP data in Table 2E.

PFam analysis indicated that the NOV2a protein contains the domains shown in Table 2F.

Example 3
The NOV3 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 3A.

Further analysis of the NOV3a protein resulted in the following properties shown in Table 3B.

A search for the NOV3a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 3C.

In a BLAST search of the public sequence database, the NOV3a protein was found to have homology with the proteins shown in the BLASTP data in Table 3D.

PFam analysis indicated that the NOV3a protein contains the domains shown in Table 3E.

Example 4
The NOV4 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 4A.

Further analysis of the NOV4a protein resulted in the following properties shown in Table 4B.

A search for the NOV4a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 4C.

In the BLAST search of the public sequence database, the NOV4a protein was found to have homology with the proteins shown in the BLASTP data in Table 4D.

PFam analysis showed that the NOV4a protein contains the domains shown in Table 4E.

Example 5
The NOV5 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 5A.

Further analysis of the NOV5a protein resulted in the following properties shown in Table 5B.

A search for the NOV5a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 5C.

In the BLAST search of the public sequence database, the NOV5a protein was found to have homology with the proteins shown in the BLASTP data in Table 5D.

PFam analysis showed that the NOV5a protein contains the domains shown in Table 5E.

Example 6
The NOV6 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 6A.

Further analysis of the NOV6a protein resulted in the following properties shown in Table 6B.

Searching for the NOV6a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 6C.

In a BLAST search of the public sequence database, the NOV6a protein was found to be homologous to the proteins shown in the BLASTP data in Table 6D.

PFam analysis indicated that the NOV6a protein contains the domains shown in Table 6E.

Example 7
The NOV7 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 7A.

Further analysis of the NOV7a protein resulted in the following properties shown in Table 7B.

A search for the NOV7a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 7C.

In a BLAST search of the public sequence database, the NOV7a protein was found to be homologous to the proteins shown in the BLASTP data in Table 7D.

PFam analysis indicated that the NOV7a protein contains the domains shown in Table 7E.

Example 8
The NOV8 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 8A.

Continuation of Table 8A

Further analysis of the NOV8a protein resulted in the following properties shown in Table 8B.

A search for the NOV8a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 8C.

In a BLAST search of the public sequence database, the NOV8a protein was found to be homologous to the proteins shown in the BLASTP data in Table 8D.

PFam analysis showed that the NOV8a protein contains the domains shown in Table 8E.

Example 9
The NOV9 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 9A.

Continuation of Table 9A

Further analysis of the NOV9a protein resulted in the following properties shown in Table 9B.

A search for the NOV9a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 9C.

In the BLAST search of the public sequence database, the NOV9a protein was found to have homology with the proteins shown in the BLASTP data in Table 9D.

PFam analysis indicated that the NOV9a protein contains the domains shown in Table 9E.

Example 10
The NOV10 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 10A.

Further analysis of the NOV10a protein resulted in the following properties shown in Table 10B.

A search for the NOV10a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 10C.

In a BLAST search of the public sequence database, the NOV10a protein was found to have homology with the proteins shown in the BLASTP data in Table 10D.

PFam analysis indicated that the NOV10a protein contains the domains shown in Table 10E.

Example 11
The NOV11 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 11A.

Further analysis of the NOV11a protein resulted in the following properties shown in Table 11B.

A search for the NOV11a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 11C.

In the BLAST search of the public sequence database, the NOV11a protein was found to have homology with the protein shown in the BLASTP data of Table 11D.

PFam analysis indicated that the NOV11a protein contains the domains shown in Table 11E.

Example 12
The NOV12 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 12A.

Further analysis of the NOV12a protein resulted in the following properties shown in Table 12B.

A search for the NOV12a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 12C.

In the BLAST search of the public sequence database, the NOV12a protein was found to have homology with the proteins shown in the BLASTP data in Table 12D.

PFam analysis indicated that the NOV12a protein contains the domains shown in Table 12E.

Example 13
The NOV13 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 13A.

Continuation of Table 13A

Continuation of Table 13A

By comparing the protein sequences, the following sequence relationships shown in Table 13B were obtained.

Further analysis of the NOV13a protein resulted in the following properties shown in Table 13C.

A search for the NOV13a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 13D.

In the BLAST search of the public sequence database, the NOV13a protein was found to have homology with the proteins shown in the BLASTP data in Table 13E.

PFam analysis indicated that the NOV13a protein contains the domains shown in Table 13F.

Example 14
The NOV14 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 14A.

By comparing the protein sequences, the following sequence relationships shown in Table 14B were obtained.

Further analysis of the NOV14a protein resulted in the following properties shown in Table 14C.

A search for the NOV14a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 14D.

In a BLAST search of the public sequence database, the NOV14a protein was found to have homology with the proteins shown in the BLASTP data in Table 14E.

PFam analysis showed that the NOV14a protein contains the domains shown in Table 14F.

Example 15
The NOV15 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 15A.

Continuation of Table 15A

By comparing the protein sequences, the following sequence relationships shown in Table 15B were obtained.

Further analysis of the NOV15a protein resulted in the following properties shown in Table 15C.

A search for the NOV15a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 15D.

In a BLAST search of the public sequence database, the NOV15a protein was found to be homologous to the proteins shown in the BLASTP data in Table 15E.

PFam analysis indicated that the NOV15a protein contains the domains shown in Table 15F.

Example 16
The NOV16 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 16A.

Continuation of Table 16A

Continuation of Table 16A

By comparing the protein sequences, the following sequence relationships shown in Table 16B were obtained.

Further analysis of the NOV16a protein resulted in the following properties shown in Table 16C.

A search for the NOV16a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 16D.

In a BLAST search of the public sequence database, the NOV16a protein was found to have homology with the protein shown in the BLASTP data of Table 16E.

PFam analysis indicated that the NOV16a protein contains the domains shown in Table 16F.

Example 17
The NOV17 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 17A.

Continuation of Table 17A

Continuation of Table 17A

By comparing the protein sequences, the following sequence relationships shown in Table 17B were obtained.

Further analysis of the NOV17a protein resulted in the following properties shown in Table 17C.

A search for the NOV17a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 17D.

In a BLAST search of the public sequence database, the NOV17a protein was found to have homology with the proteins shown in the BLASTP data of Table 17E.

PFam analysis indicated that the NOV17a protein contains the domains shown in Table 17F.

Example 18
The NOV18 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 18A.

Continuation of Table 18A

Further analysis of the NOV18a protein resulted in the following properties shown in Table 18B.

A search for the NOV18a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 18C.

In a BLAST search of the public sequence database, the NOV18a protein was found to have homology with the proteins shown in the BLASTP data in Table 18D.

PFam analysis indicated that the NOV18a protein contains the domains shown in Table 18E.

Example 19
The NOV19 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 19A.

Continuation of Table 19A

Further analysis of the NOV19a protein resulted in the following properties shown in Table 19B.

A search for the NOV19a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 19C.

In the BLAST search of the public sequence database, the NOV19a protein was found to have homology with the protein shown in the BLASTP data of Table 19D.

PFam analysis indicated that the NOV19a protein contains the domains shown in Table 19E.

Example 20
The NOV20 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 20A.

Continuation of Table 20A

Continuation of Table 20A

By comparing the protein sequences, the following sequence relationships shown in Table 20B were obtained.

Further analysis of the NOV20a protein resulted in the following properties shown in Table 20C.

A search for the NOV20a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 20D.

In the BLAST search of the public sequence database, the NOV20a protein was found to have homology with the proteins shown in the BLASTP data of Table 20E.

PFam analysis indicated that the NOV20a protein contains the domains shown in Table 20F.

Example 21
The NOV21 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 21A.

Further analysis of the NOV21a protein resulted in the following properties shown in Table 21B.

A search for the NOV21a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 21C.

In the BLAST search of the public sequence database, the NOV21a protein was found to have homology with the protein shown in the BLASTP data in Table 21D.

PFam analysis indicated that the NOV21a protein contains the domains shown in Table 21E.

Example 22
The NOV22 clone was analyzed. Polypeptide sequences encoded by nucleotides are shown in Table 22A.

Continuation of Table 22A

Further analysis of the NOV22a protein resulted in the following properties shown in Table 22B.

A search for the NOV22a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 22C.

In a BLAST search of the public sequence database, the NOV22a protein was found to have homology with the proteins shown in the BLASTP data in Table 22D.

PFam analysis indicated that the NOV22a protein contains the domains shown in Table 22E.

Example 23
The NOV23 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 23A.

Continuation of Table 23A

Continuation of Table 23A

Continuation of Table 23A

Continuation of Table 23A

Continuation of Table 23A

By comparing the protein sequences, the following sequence relationships shown in Table 23B were obtained.

Further analysis of the NOV23a protein resulted in the following properties shown in Table 23C.

A search for the NOV23a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 23D.

In the BLAST search of the public sequence database, the NOV23a protein was found to have homology with the proteins shown in the BLASTP data of Table 23E.

PFam analysis indicated that the NOV23a protein contains the domains shown in Table 23F.

Example 24
The NOV24 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 24A.

Further analysis of the NOV24a protein resulted in the following properties shown in Table 24B.

A search for the NOV24a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 24C.

In the BLAST search of the public sequence database, the NOV24a protein was found to have homology with the proteins shown in the BLASTP data in Table 24D.

PFam analysis indicated that the NOV24a protein contains the domains shown in Table 24E.

Example 25
The NOV25 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 25A.

Continuation of Table 25A

Continuation of Table 25A

Continuation of Table 25A

Continuation of Table 25A

By comparing the protein sequences, the following sequence relationships shown in Table 25B were obtained.

Further analysis of the NOV25a protein resulted in the following properties shown in Table 25C.

A search for the NOV25a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 25D.

In a BLAST search of the public sequence database, the NOV25a protein was found to have homology with the proteins shown in the BLASTP data in Table 25E.

PFam analysis indicated that the NOV25a protein contains the domains shown in Table 25F.

Example 26
The NOV26 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 26A.

Continuation of Table 26A

By comparing the protein sequences, the following sequence relationships shown in Table 26B were obtained.

Further analysis of the NOV26a protein resulted in the following properties shown in Table 26C.

A search for the NOV26a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 26D.

In a BLAST search of the public sequence database, the NOV26a protein was found to be homologous to the protein shown in the BLASTP data in Table 26E.

PFam analysis indicated that the NOV26a protein contains the domains shown in Table 26F.

Example 27
The NOV27 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 27A.

Further analysis of the NOV27a protein resulted in the following properties shown in Table 27B.

A search for the NOV27a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 27C.

In a BLAST search of the public sequence database, the NOV27a protein was found to have homology with the protein shown in the BLASTP data in Table 27D.

PFam analysis indicated that the NOV27a protein contains the domains shown in Table 27E.

Example 28
The NOV28 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 28A.

Continuation of Table 28A

Continuation of Table 28A

By comparing the protein sequences, the following sequence relationships shown in Table 28B were obtained.

Further analysis of the NOV28a protein resulted in the following properties shown in Table 28C.

A search for the NOV28a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 28D.

In a BLAST search of the public sequence database, the NOV28a protein was found to be homologous to the protein shown in the BLASTP data in Table 28E.

PFam analysis indicated that the NOV28a protein contains the domains shown in Table 28F.

Example 29
The NOV29 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 29A.

By comparing the protein sequences, the following sequence relationships shown in Table 29B were obtained.

Further analysis of the NOV29a protein resulted in the following properties shown in Table 29C.

A search for the NOV29a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 29D.

In a BLAST search of the public sequence database, the NOV29a protein was found to have homology with the protein shown in the BLASTP data in Table 29E.

PFam analysis showed that the NOV29a protein contains the domains shown in Table 29F.

Example 30
The NOV30 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 30A.

Continuation of Table 30A

Continuation of Table 30A

Further analysis of the NOV30a protein resulted in the following properties shown in Table 30B.

A search for the NOV30a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 30C.

In a BLAST search of the public sequence database, the NOV30a protein was found to have homology with the proteins shown in the BLASTP data in Table 30D.

PFam analysis indicated that the NOV30a protein contains the domains shown in Table 30E.

Example 31
The NOV31 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 31A.

Continuation of Table 31A

By comparing the protein sequences, the following sequence relationships shown in Table 31B were obtained.

Further analysis of the NOV31a protein resulted in the following properties shown in Table 31C.

A search for the NOV31a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 31D.

In the BLAST search of the public sequence database, the NOV31a protein was found to have homology with the proteins shown in the BLASTP data of Table 31E.

PFam analysis indicated that the NOV31a protein contains the domains shown in Table 31F.

Example 32
The NOV32 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 32A.

Continuation of Table 32A

By comparing the protein sequences, the following sequence relationships shown in Table 32B were obtained.

Further analysis of the NOV32a protein resulted in the following properties shown in Table 32C.

A search for the NOV32a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 32D.

In a BLAST search of the public sequence database, the NOV32a protein was found to have homology with the proteins shown in the BLASTP data in Table 32E.

PFam analysis indicated that the NOV32a protein contains the domains shown in Table 32F.

Example 33
The NOV33 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 33A.

Further analysis of the NOV33a protein resulted in the following properties shown in Table 33B.

A search for the NOV33a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 33C.

In a BLAST search of the public sequence database, the NOV33a protein was found to have homology with the proteins shown in the BLASTP data in Table 33D.

PFam analysis indicated that the NOV33a protein contains the domains shown in Table 33E.

Example 34
The NOV34 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 34A.

Continuation of Table 34A

By comparing the protein sequences, the following sequence relationships shown in Table 34B were obtained.

Further analysis of the NOV34a protein resulted in the following properties shown in Table 34C.

A search for the NOV34a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 34D.

In a BLAST search of the public sequence database, the NOV34a protein was found to be homologous to the proteins shown in the BLASTP data in Table 34E.

PFam analysis indicated that the NOV34a protein contains the domains shown in Table 34F.

Example 35
The NOV35 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 35A.

Further analysis of the NOV35a protein resulted in the following properties shown in Table 35B.

A search for the NOV35a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 35C.

In a BLAST search of the public sequence database, the NOV35a protein was found to be homologous to the protein shown in the BLASTP data in Table 35D.

PFam analysis indicated that the NOV35a protein contains the domains shown in Table 35E.

Example 36
The NOV36 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 36A.

By comparing the protein sequences, the following sequence relationships shown in Table 36B were obtained.

Further analysis of the NOV36a protein resulted in the following properties shown in Table 36C.

A search for the NOV36a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 36D.

In a BLAST search of the public sequence database, the NOV36a protein was found to have homology with the proteins shown in the BLASTP data in Table 36E.

PFam analysis indicated that the NOV36a protein contains the domains shown in Table 36F.

Example 37
The NOV37 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 37A.

Continuation of Table 37A

Further analysis of the NOV37a protein resulted in the following properties shown in Table 37B.

A search for the NOV37a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 37C.

In a BLAST search of the public sequence database, the NOV37a protein was found to have homology with the proteins shown in the BLASTP data in Table 37D.

PFam analysis indicated that the NOV37a protein contains the domains shown in Table 37E.

Example 38
The NOV38 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 38A.

Further analysis of the NOV38a protein resulted in the following properties shown in Table 38B.

A search for the NOV38a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 38C.

In a BLAST search of the public sequence database, the NOV38a protein was found to be homologous to the proteins shown in the BLASTP data in Table 38D.

PFam analysis indicated that the NOV38a protein contains the domains shown in Table 38E.

Example 39
The NOV39 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 39A.

Continuation of Table 39A

Further analysis of the NOV39a protein resulted in the following properties shown in Table 39B.

A search for the NOV39a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 39C.

In a BLAST search of the public sequence database, the NOV39a protein was found to have homology with the proteins shown in the BLASTP data in Table 39D.

PFam analysis showed that the NOV39a protein contains the domains shown in Table 39E.

Example 40
The NOV40 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 40A.

Continuation of Table 40A

Further analysis of the NOV40a protein resulted in the following properties shown in Table 40B.

A search for the NOV40a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 40C.

In the BLAST search of the public sequence database, the NOV40a protein was found to have homology with the protein shown in the BLASTP data of Table 40D.

PFam analysis showed that the NOV40a protein contains the domains shown in Table 40E.

Example 41
The NOV41 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 41A.

Further analysis of the NOV41a protein resulted in the following properties shown in Table 41B.

A search for the NOV41a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 41C.

In the BLAST search of the public sequence database, the NOV41a protein was found to have homology with the proteins shown in the BLASTP data of Table 41D.

PFam analysis showed that the NOV41a protein contains the domains shown in Table 41E.

Example 42
The NOV42 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 42A.

Further analysis of the NOV42a protein resulted in the following properties shown in Table 42B.

A search for the NOV42a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 42C.

In a BLAST search of the public sequence database, the NOV42a protein was found to have homology with the proteins shown in the BLASTP data in Table 42D.

PFam analysis showed that the NOV42a protein contains the domains shown in Table 42E.

Example 43
The NOV43 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 43A.

Further analysis of the NOV43a protein resulted in the following properties shown in Table 43B.

A search for the NOV43a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 43C.

In the BLAST search of the public sequence database, the NOV43a protein was found to have homology with the protein shown in the BLASTP data in Table 43D.

PFam analysis showed that the NOV43a protein contains the domains shown in Table 43E.

Example 44
The NOV44 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 44A.

Further analysis of the NOV44a protein resulted in the following properties shown in Table 44B.

A search for the NOV44a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 44C.

In a BLAST search of the public sequence database, the NOV44a protein was found to have homology with the proteins shown in the BLASTP data in Table 44D.

PFam analysis indicated that the NOV44a protein contains the domains shown in Table 44E.

Example 45
The NOV45 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 45A.

Continuation of Table 45A

Further analysis of the NOV54a protein resulted in the following properties shown in Table 45B.

A search for the NOV45a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 45C.

In a BLAST search of the public sequence database, the NOV45a protein was found to have homology with the protein shown in the BLASTP data in Table 45D.

PFam analysis indicated that the NOV45a protein contains the domains shown in Table 45E.

Example 46
The NOV46 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 46A.

By comparing the protein sequences, the following sequence relationships shown in Table 46B were obtained.

Further analysis of the NOV46a protein resulted in the following properties shown in Table 46C.

A search for the NOV46a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 46D.

In a BLAST search of the public sequence database, the NOV46a protein was found to have homology with the protein shown in the BLASTP data in Table 46E.

PFam analysis indicated that the NOV46a protein contains the domains shown in Table 46F.

Example 47
The NOV47 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 47A.

Further analysis of the NOV47a protein resulted in the following properties shown in Table 47B.

A search for the NOV47a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 47C.

In the BLAST search of the public sequence database, the NOV47a protein was found to have homology with the protein shown in the BLASTP data in Table 47D.

PFam analysis indicated that the NOV47a protein contains the domains shown in Table 47E.

Example 48
The NOV48 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 48A.

Continuation of Table 48A

Further analysis of the NOV48a protein resulted in the following properties shown in Table 48B.

A search for the NOV48a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 48C.

In a BLAST search of the public sequence database, the NOV48a protein was found to have homology with the protein shown in the BLASTP data in Table 48D.

PFam analysis indicated that the NOV48a protein contains the domains shown in Table 48E.

Example 49
The NOV49 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 49A.

Further analysis of the NOV49a protein resulted in the following properties shown in Table 49B.

A search for the NOV49a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 49C.

In a BLAST search of the public sequence database, the NOV49a protein was found to have homology with the protein shown in the BLASTP data in Table 49D.

PFam analysis showed that the NOV49a protein contains the domains shown in Table 49E.

Example 50
The NOV50 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 50A.

Continuation of Table 50A

By comparing the protein sequences, the following sequence relationships shown in Table 50B were obtained.

Further analysis of the NOV50a protein resulted in the following properties shown in Table 50C.

A search for the NOV50a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 50D.

In a BLAST search of the public sequence database, the NOV50a protein was found to be homologous to the proteins shown in the BLASTP data in Table 50E.

PFam analysis showed that the NOV50a protein contains the domains shown in Table 50F.

Example 51
The NOV51 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 51A.

Further analysis of the NOV51a protein resulted in the following properties shown in Table 51B.

A search for the NOV51a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 51C.

In the BLAST search of the public sequence database, the NOV51a protein was found to have homology with the protein shown in the BLASTP data of Table 51D.

PFam analysis indicated that the NOV51a protein contains the domains shown in Table 51E.

Example 52
The NOV52 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 52A.

Continuation of Table 52A

Further analysis of the NOV52a protein resulted in the following properties shown in Table 52B.

A search for the NOV52a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 52C.

In the BLAST search of the public sequence database, the NOV52a protein was found to have homology with the proteins shown in the BLASTP data in Table 52D.

PFam analysis indicated that the NOV52a protein contains the domains shown in Table 52E.

Example 53
The NOV53 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 53A.

Continuation of Table 53A

Further analysis of the NOV53a protein resulted in the following properties shown in Table 53B.

A search for the NOV53a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 53C.

In the BLAST search of the public sequence database, the NOV53a protein was found to have homology with the protein shown in the BLASTP data of Table 53D.

PFam analysis indicated that the NOV53a protein contains the domains shown in Table 53E.

Example 54
The NOV54 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 54A.

Continuation of Table 54A

Further analysis of the NOV54a protein resulted in the following properties shown in Table 54B.

A search for the NOV54a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 54C.

In a BLAST search of the public sequence database, the NOV54a protein was found to have homology with the proteins shown in the BLASTP data in Table 54D.

PFam analysis indicated that the NOV54a protein contains the domains shown in Table 54E.

Example 55
The NOV55 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 55A.

Continuation of Table 55A

Continuation of Table 55A

By comparing the protein sequences, the following sequence relationships shown in Table 55B were obtained.

Further analysis of the NOV55a protein resulted in the following properties shown in Table 55C.

A search for the NOV55a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 55D.

In a BLAST search of the public sequence database, the NOV55a protein was found to have homology with the protein shown in the BLASTP data in Table 55E.

PFam analysis indicated that the NOV55a protein contains the domains shown in Table 55F.

Example 56
The NOV56 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 56A.

Further analysis of the NOV56a protein resulted in the following properties shown in Table 56B.

A search for the NOV56a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 56C.

In the BLAST search of the public sequence database, the NOV56a protein was found to have homology with the protein shown in the BLASTP data of Table 56D.

PFam analysis indicated that the NOV56a protein contains the domains shown in Table 56E.

Example 57
The NOV57 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 57A.

Further analysis of the NOV57a protein resulted in the following properties shown in Table 57B.

A search for the NOV57a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 57C.

In the BLAST search of the public sequence database, the NOV57a protein was found to have homology with the protein shown in the BLASTP data in Table 57D.

PFam analysis showed that the NOV57a protein contains the domains shown in Table 57E.

Example 58
The NOV58 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 58A.

Further analysis of the NOV58a protein resulted in the following properties shown in Table 58B.

A search for the NOV58a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 58C.

In a BLAST search of the public sequence database, the NOV58a protein was found to have homology with the protein shown in the BLASTP data in Table 58D.

PFam analysis indicated that the NOV58a protein contains the domains shown in Table 58E.

Example 59
The NOV59 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 59A.

Continuation of Table 59A

Further analysis of the NOV59a protein resulted in the following properties shown in Table 59B.

A search for the NOV59a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 59C.

In the BLAST search of the public sequence database, the NOV59a protein was found to have homology with the protein shown in the BLASTP data in Table 59D.

PFam analysis showed that the NOV59a protein contains the domains shown in Table 59E.

Example 60
The NOV60 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 60A.

Further analysis of the NOV60a protein resulted in the following properties shown in Table 60B.

A search for the NOV60a protein in the Geneseq database, which is a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 60C.

In a BLAST search of the public sequence database, the NOV60a protein was found to have homology with the proteins shown in the BLASTP data of Table 60D.

PFam analysis indicated that the NOV60a protein contains the domains shown in Table 60E.

Example 61
The NOV61 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 61A.

Continuation of Table 61A

By comparing the protein sequences, the following sequence relationships shown in Table 61B were obtained.

Further analysis of the NOV61a protein resulted in the following properties shown in Table 61C.

A search for the NOV61a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 61D.

In the BLAST search of the public sequence database, the NOV61a protein was found to have homology with the protein shown in the BLASTP data in Table 61E.

PFam analysis indicated that the NOV61a protein contains the domains shown in Table 61F.

Example 62
The NOV62 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 62A.

By comparing the protein sequences, the following sequence relationships shown in Table 62B were obtained.

Further analysis of the NOV62a protein resulted in the following properties shown in Table 62C.

A search for the NOV62a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, resulted in several homologous proteins shown in Table 62D.

In a BLAST search of the public sequence database, the NOV62a protein was found to have homology with the proteins shown in the BLASTP data in Table 62E.

PFam analysis indicated that the NOV62a protein contains the domains shown in Table 62F.

Example 63
The NOV63 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 63A.

Continuation of Table 63A

Further analysis of the NOV63a protein resulted in the following properties shown in Table 63B.

A search for the NOV63a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 63C.

In the BLAST search of the public sequence database, the NOV63a protein was found to have homology with the protein shown in the BLASTP data in Table 63D.

PFam analysis indicated that the NOV63a protein contains the domains shown in Table 63E.

Example 64
The NOV64 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 64A.

Table 64A continued

Further analysis of the NOV64a protein resulted in the following properties shown in Table 64B.

A search for the NOV64a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 64C.

In a BLAST search of the public sequence database, the NOV64a protein was found to be homologous to the proteins shown in the BLASTP data in Table 64D.

PFam analysis indicated that the NOV64a protein contains the domains shown in Table 64E.

Example 65
The NOV65 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 65A.

By comparing the protein sequences, the following sequence relationships shown in Table 65B were obtained.

Further analysis of the NOV65a protein resulted in the following properties shown in Table 65C.

A search for the NOV65a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 65D.

In a BLAST search of the public sequence database, the NOV65a protein was found to be homologous to the proteins shown in the BLASTP data in Table 65E.

PFam analysis indicated that the NOV65a protein contains the domains shown in Table 65F.

Example 66
The NOV66 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 66A.

Further analysis of the NOV66a protein resulted in the following properties shown in Table 66B.

A search for the NOV66a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 66C.

In a BLAST search of the public sequence database, the NOV66a protein was found to have homology with the proteins shown in the BLASTP data in Table 66D.

PFam analysis indicated that the NOV66a protein contains the domains shown in Table 66E.

Example 67
The NOV67 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 67A.

Further analysis of the NOV67a protein resulted in the following properties shown in Table 67B.

A search for the NOV67a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 67C.

In the BLAST search of the public sequence database, the NOV67a protein was found to have homology with the protein shown in the BLASTP data in Table 67D.

PFam analysis showed that the NOV67a protein contains the domains shown in Table 67E.

Example 68
The NOV68 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 68A.

Continuation of Table 68A

By comparing the protein sequences, the following sequence relationships shown in Table 68B were obtained.

Further analysis of the NOV68a protein resulted in the following properties shown in Table 68C.

A search for the NOV68a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 68D.

In a BLAST search of the public sequence database, the NOV68a protein was found to have homology with the protein shown in the BLASTP data in Table 68E.

PFam analysis indicated that the NOV68a protein contains the domains shown in Table 68F.

Example 69
The NOV69 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 69A.

Further analysis of the NOV69a protein resulted in the following properties shown in Table 69B.

A search for the NOV69a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 69C.

In a BLAST search of the public sequence database, the NOV69a protein was found to have homology with the proteins shown in the BLASTP data in Table 69D.

PFam analysis showed that the NOV69a protein contains the domains shown in Table 69E.

Example 70
The NOV70 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 70A.

Continuation of Table 70A

Continuation of Table 70A

By comparing the protein sequences, the following sequence relationships shown in Table 70B were obtained.

Further analysis of the NOV70a protein resulted in the following properties shown in Table 70C.

A search for the NOV70a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 70D.

In the BLAST search of the public sequence database, the NOV70a protein was found to have homology with the proteins shown in the BLASTP data in Table 70E-1.

In the BLAST search of the public sequence database, the NOV70e protein was found to have homology with the proteins shown in the BLASTP data in Table 70E-2.

PFam analysis indicated that the NOV70a protein contains the domains shown in Table 70F.

Example 71
The NOV71 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 71A.

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

Continuation of Table 71A

By comparing the protein sequences, the following sequence relationships shown in Table 71B were obtained.

Further analysis of the NOV71a protein resulted in the following properties shown in Table 71C.

A search for the NOV71a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 71D.

In the BLAST search of the public sequence database, the NOV71a protein was found to have homology with the proteins shown in the BLASTP data in Table 71E.

PFam analysis indicated that the NOV71a protein contains the domains shown in Table 71F.

Example 72
The NOV72 clone was analyzed. The polypeptide sequences encoded by the nucleotides are shown in Table 72A.

Further analysis of the NOV72a protein resulted in the following properties shown in Table 72B.

A search for the NOV72a protein in the Geneseq database, a proprietary database containing sequences published in patents and patent publications, yielded several homologous proteins shown in Table 72C.

In a BLAST search of the public sequence database, the NOV72a protein was found to have homology with the proteins shown in the BLASTP data in Table 72D.

PFam analysis indicated that the NOV72a protein contains the domains shown in Table 72E.

Example B: Sequencing Methodology and Identification of NOVX Clones The following techniques were used in the examples of the present invention.
1. GeneCalling ™ technique: This is a proprietary method for performing differential gene expression profiling between two or more samples developed at CuraGen, Shimkets, et al., “Gene expression analysis by transcript profiling coupled to a gene database query "Nature Biotechnology 17: 198-803 (1999). cDNA was derived from various human samples representing multiple tissue types, normal and pathological conditions, physiological conditions and developmental conditions from different donors. Samples were obtained as whole tissue, primary cells or tissue culture primary cells or cell lines. Cells and cell lines may be treated with biological or chemical agents that modulate gene expression, such as growth factors, chemokines or steroids. The cDNA thus derived was then digested with as many as 120 pairs of restriction enzymes, and linker-adapter pairs specific for each pair of restriction enzymes were ligated to the appropriate ends. Restriction digests produce a mixture of unique cDNA gene fragments. Limited PCR amplification is performed with primers that are homologous to a linker adapter sequence in which one primer is biotinylated and the other is fluorescently labeled. The doubly labeled material was isolated and the fluorescently labeled single strand was resolved by capillary gel electrophoresis. A computer algorithm compares the experiments from each restriction digest and electrophoresis from the control group. Information from this and further sequences is used to predict the identity of each differentially expressed gene fragment using various genomic databases. The identity of the gene fragment was confirmed by additional, gene-specific competitive PCR or by gene fragment isolation and sequencing.

  2. SeqCalling® technology: cDNA was derived from various human samples representing multiple tissue types, normal and pathological conditions, physiological conditions and developmental conditions from different donors. Samples were obtained as whole tissue, primary cells or tissue culture primary cells or cell lines. Cells and cell lines may be treated with biological or chemical agents that modulate gene expression, such as growth factors, chemokines or steroids. The cDNA thus derived was then sequenced using CuraGen's proprietary SeqCalling technology. Sequence traces were manually evaluated and edited for correction if appropriate. The cDNA sequences from all samples were collected together using a bioinformatic program that sometimes included public human sequences to produce a common sequence for each assembly. Each assembly is included in the Curagen Corporation database. A sequence was included as a component of an assembly when the degree of identity with other components was at least 95% over 50 bp. Each assembly represents a gene or part thereof and contains information about variants such as splice forms of single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations.

3. PathCalling technology:
NOVX nucleic acid sequences are derived by laboratory screening of cDNA libraries by a two-hybrid approach. A cDNA fragment that covers either the full length of the DNA sequence or part of the sequence or both was sequenced. In silico predictions were based on sequences available in Curagen Corporation's proprietary sequence database or public human sequence database and provided either the full-length DNA sequence or some portion thereof.

Laboratory screening was performed using the method summarized below:
The cDNA library was derived from various human samples representing multiple tissue types, normal and pathological conditions, physiological conditions and developmental conditions from different donors. Samples were obtained as whole tissue, primary cells or tissue culture primary cells or cell lines. Cells and cell lines may be treated with biological or chemical agents that modulate gene expression, such as growth factors, chemokines or steroids. The cDNA thus derived was then directionally cloned into an appropriate two hybrid vector (Gal4-activation domain (Gal4-AD) fusion). Such cDNA libraries as well as commercially available cDNA libraries from Clontech (Palo Alto, Calif.), And then the proprietary yeast strains of Curagen Corporation from E. coli (US Pat. Nos. 6,057,101 and 6,083). , 693 (disclosed in the entire specification as a part of this specification).

  Multiple Gal4-AD fusion cDNA libraries were screened using Gal4-binding domain (Gal4-BD) fusions from Curagen Corporation's proprietary human sequence library, with each Gal4-AD fusion containing a separate cDNA. This resulted in selection of yeast hybrid diploid containing. Each sample was amplified using polymerase chain reaction (PCR) with non-specific primers in the cDNA insertion boundary. Such PCR products were sequenced; sequence traces were manually evaluated and edited for correction if appropriate. Using bioinformatic programs, sometimes including public human sequences, cDNA sequences from all samples were collected together to produce a common sequence for each assembly. Each assembly is included in the Curagen Corporation database. A sequence was included as a component of the assembly when the degree of identity with other components was at least 95% over 50 bp. Each assembly represents a gene or portion thereof and includes information about variants such as splice forms of single nucleotide polymorphisms (SNPs), insertions, deletions and other sequence variations.

  Physical clone: A cDNA fragment derived by a screening procedure covering all open reading frames was cloned into the pACT2 plasmid (Clontech) used as recombinant DNA to create a cDNA library. The recombinant plasmid was inserted into the host and produced during the screening procedure by crossing both Curagen Corporation's proprietary yeast strains N106 ′ and YULH (US Pat. Nos. 6,057,101 and 6,083,693). Selected by diploid.

  4). RACE: Polymerase chain reaction based techniques such as rapid amplification of cDNA ends (RACE) were used to isolate or complete the predicted sequence of the cDNA of the present invention. Usually, multiple clones were sequenced from one or more human samples to derive the sequence for the fragment. Various human tissue samples from different donors were used for the RACE reaction. Sequences derived from these procedures were included in the SeqCalling Assembly method described in the previous chapter.

  5). Exon ligation: The NOVX target sequence identified in the present invention was subjected to the exon ligation method to confirm the sequence. PCR primers were designed starting from the most upstream sequence available for the forward primer and from the most downstream sequence available for the reverse primer. In each case, walk inward from each end to the coding sequence until an appropriate sequence that is either unique or highly selective is encountered, or in the case of a reverse primer, a stop codon is reached. And examined the sequence. Such primers are based on in silico predictions for full-length cDNA, target sequence DNA or protein sequence portions (one or more exons), or predicted exons for closely related human sequences from other species. Designed with the translated homology of These primers were then used in PCR amplification based on the following human cDNA pools: adrenal gland, bone marrow, brain-tonsillar, brain-cerebellum, brain-hippocampus, brain-substantia nigra, brain-thalamus, brain-full margin, Fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma-large, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea The womb. Usually, the resulting amplicons were gel purified, cloned, and sequenced to high redundancy. PCR products derived from exon binding were cloned into Invitrogen's pCR2.1 vector. The resulting bacterial clone has an insert that covers the complete open reading frame cloned into the pCR2.1 vector. Sequences from all clones were assembled together with themselves, other fragments in the CuraGen Corporation database, and public ESTs. Fragments and ESTs were included as components of the assembly when their degree of identity with other components of the assembly was at least 95% over 50 bp. In addition, sequence traces were manually evaluated and edited for correction if appropriate. These procedures provide the sequences reported herein.

6). Physical clones: Exons were predicted by homology, and intron / exon boundaries were determined using standard genetic rules. Exons were further selected and refined by multiple BLAST (eg, tBlastN, BlastX and BlastN) and, in some cases, similarity measures using GeneScan and Grail. When available, expression sequences from both public and proprietary databases were also added to further define and complete the gene sequences. The DNA sequence was then manually corrected for obvious discrepancies, thereby obtaining the sequence encoding the full-length protein.
PCR products derived by exon binding covering all open reading frames were cloned into Invitrogen's pCR2.1 vector to provide clones for expression and screening purposes.

Example C: Quantitative expression analysis of clones in various cells and tissues Real-time quantitative PCR (RTQ PCR) using microtiter plates containing RNA samples from cells, cell lines, and tissues from various normal and lesions Was used to evaluate the quantitative expression of various clones. RTQ PCR was performed on an Applied Biosystems ABI PRISM® 7700 or ABI PRISM® 7900 HT Sequence Detection System. Various collected samples are collected on the plate, panel 1 (containing normal tissues and cancer cell lines), panel 2 (containing samples from tissues of normal and cancer origin), panel 3 (cancer cell lines). Panel 4 (contains cells and cell lines from normal tissues and cells associated with inflammatory abnormalities), panel 5D / 5I (contains human tissues and cell lines highlighting metabolic disease), AI _Comprehensive_panel (containing samples from normal tissue and autoimmune disease), panel CNSD.01 (containing central nervous system samples from normal and diseased brain), and CNS_neurodegeneration_panel (normal and Containing samples from Alzheimer's disease brain).

  By agarose gel electropherogram using 28S and 18S ribosomal RNA staining intensity ratio as a guide (2: 1-2.5: 1 28s: 18s) and by visual assessment of absence of low molecular weight RNA that would indicate degradation products Quality control the integrity of RNA from all samples. Samples are controlled against genomic DNA contamination by RTQ PCR reactions performed in the absence of reverse transcriptase using a set of probes and primers designed to amplify over a single exon interval.

  Initially, RNA samples were normalized to reference nucleic acids such as constitutively expressed genes (eg, β-actin and GAPDH). Normalized RNA (5 μl) is converted to cDNA and analyzed by RTQ PCR using One Step RT-PCR Master Mix Reagents (Applied Biosystems; Catalog No. 4309169) and gene-specific primers according to manufacturer's instructions did.

  In other cases, non-standardized RNA samples are made into single-stranded cDNA (sscDNA) using Superscript II (Invitrogen Corporation; Catalog No. 18064-147) and disordered hexamers according to the manufacturer's instructions. Converted. Reactions containing up to 10 μg total RNA were performed in a 20 μl volume and incubated at 42 ° C. for 60 minutes. This reaction can be scaled up to 50 μg total RNA in a final volume of 100 μl. The sscDNA sample is then normalized to the reference nucleic acid as described above using 1X TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020) according to the manufacturer's instructions.

  Probes and primers were designed for each assay according to the Applied Biosystems Primer Express Software package (version 1 for Apple Computer's Macintosh Power PC) or a similar algorithm using the target sequence as input. The default settings were used for the reaction conditions, the following parameters were set, and primers were selected: primer concentration = 250 nM, primer melting temperature (Tm) range = 58 ° -60 ° C., primer optimal Tm = 59 ° C., maximum Primer difference = 2 ° C., probe does not have 5′G, probe Tm must be 10 ° C. larger than primer Tm, amplicon (amplification product) size is 75 bp to 100 bp. Selected probes and primers (see below) were synthesized by Synthegen (Houston, TX, USA). The probe was purified twice by HPLC to remove uncoupled dye and evaluated by mass spectrometry to confirm that the reporter and quencher dyes were coupled to the 5 'and 3' ends of the probe, respectively. Proven. Their final concentrations were 900 nM for the forward and reverse primers, respectively, and 200 nM for the probe.

  PCR conditions: When working with RNA samples, normalized RNA from each tissue and each cell line is placed in each well of either 96-well or 384-well PCR plates (Applied Biosystems). Spotted. PCR cocktails include a single gene-specific probe and primer set or two multiplex probes and primer sets (a set specific to the target clone and another gene-specific set multiplexed by the target probe). ) Is included. The PCR reaction was set up using TaqMan® One-Step RT-PCR Master Mix (Applied Biosystems; Catalog No. 4313803) according to the manufacturer's instructions. Reverse transcription was performed at 48 ° C. for 30 minutes followed by an amplification / PCR cycle as follows: 95 ° C. for 10 minutes, then 95 ° C. for 15 seconds, 60 ° C. for 1 minute, 40 cycles. The result is recorded as a CT value (cycle in which a given sample crosses the fluorescence threshold) using a log scale, and between the given sample and the sample with the lowest CT value expressed as a delta CT power of 2 The difference was the RNA concentration. Percent relative expression is then obtained by taking the reciprocal of this RNA difference and multiplying by 100.

  When working with sscDNA samples, standardized sscDNA was used as previously described for RNA samples. A PCR reaction containing one or two sets of probes and primers was set up using 1X TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020) according to the manufacturer's instructions. PCR amplification was performed as follows: 40 cycles of 95 ° C. for 10 minutes, then 95 ° C. for 15 seconds, 60 ° C. for 1 minute. Results were analyzed and processed as described above.

Panel 1, 1.1, 1.2, and 1.3D
Plates for panels 1, 1.1, 1.2, and 1.3D include two control holes (genomic DNA control and chemical control) and 94 holes containing cDNA from various samples. The samples in these panels are divided into two classes: samples from cultured cell lines and samples from primary normal tissues. Cell lines are derived from the following types of cancer: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, kidney Cancer, stomach cancer and pancreatic cancer. The cell lines used in these panels were widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using conditions recommended by the ATCC. The normal tissue seen in these panels consists of samples from all major organ systems of a single adult individual or fetus. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions of the brain, spleen , Bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and fat.

In the results for panels 1, 1.1, 1.2, and 1.3D, the following abbreviations are used.
ca. : Cancer
^* : Established from metastasis met: metastasis s cell var: small cell mutant non-s = non-sm: not small squam: flat
pl.eff = pl effusion: pleural effusion glio: glioma astro: astrocytoma neuro: neuroblastoma

General_Screening_Panel_v1.4 and General_Screening_Panel_v1.5
The plates for panels 1.4 and 1.5 contain two control holes (genomic DNA control and chemical control) and 94 holes containing cDNA from various samples. The samples in panels 1.4 and 1.5 are divided into two classes: samples from cultured cell lines and samples from primary normal tissues. Cell lines are derived from the following types of cancer: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, kidney Cancer, stomach cancer and pancreatic cancer. The cell lines used in panels 1.4 and 1.5 were cultured using the conditions recommended by the ATCC, widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines. The normal tissue seen in panel 1.4 consists of sample pools derived from all major organ systems of 2-5 different adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions of the brain, spleen , Bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, uterus, placenta, prostate, testis and fat. Abbreviations are as described for panels 1, 1.1, 1.2, and 1.3D.

Panel 2D and 2.2
Plates for panels 2D and 2.2 are typically created by surgeons working closely with two control holes and the National Cancer Institute's Cooperative Human Tissue Network (CHTN) or the National Disease Research Initiative (NDRI) Contains 94 test samples consisting of RNA or cDNA isolated from procured human tissue. Tissues are derived from human malignancies, and when indicated, many malignant tumor tissues have “corresponding margins” obtained from non-cancerous tissue immediately adjacent to the tumor. These are referred to as normal adjacent tissues, and are expressed as “NAT” in the following results. Tumor tissue and “corresponding margins” are evaluated by two independent pathologists (surgical pathologists plus NDRI or CHTN pathologists). This analysis provides a histopathological assessment of the grade of tumor differentiation. In addition, most samples contain original surgical pathology reports that provide information about the clinical stage of the patient. These matched perimeters are taken from the tissue surrounding the surgical area (ie, directly adjacent) (in Table RR, labeled “NAT” for normal adjacent tissue). In addition, RNA and cDNA samples were obtained from various human tissues derived from autopsy performed in elderly or sudden death victims (accidents, etc.). These tissues were confirmed to be disease free and were purchased from various commercial suppliers such as Clontech (Palo Alto, CA), Research Genetics and Invitrogen.

Panel 3D
Panel 3D plate consists of 94 cDNA samples and 2 control samples. In particular, 92 of these samples are derived from cultured human cancer cell lines, 2 human primary cerebellar tissue samples, and 2 controls. Human cell lines are generally obtained from the American Type Culture Collection (ATCC), NCI or the German Cancer Cell Bank and fall into the following tissue groups: squamous cell carcinoma of the tongue, breast cancer, prostate cancer, black Carcinoma, epidermoid carcinoma, sarcoma, bladder carcinoma, pancreatic cancer, kidney cancer, leukemia / lymphoma, ovary / uterus / cervix, stomach, colon, lung and CNS cancer cell lines. In addition, there are two independent cerebellar samples. All these cells are cultured under standard recommended conditions and RNA is extracted using standard techniques. The cell lines in panels 3D and 1.3D are the most common cell lines used in the scientific literature.

Panels 4D, 4R, and 4.1D
Panel 4 is a 96-well plate (2 control wells, 94, consisting of RNA (panel 4R) or cDNA (panel 4D / 4.1D) isolated from various human cell lines or tissues associated with inflammatory diseases. Samples on a single test sample). Total RNA from control normal tissues such as colon and lung (Stratagene, La Jolla, CA) and thymus and kidney (Clonetech) were used. Total RNA from liver tissue of cirrhotic patients and kidney of erythematosus patients was obtained from BioChain (Biochain Institute, Inc., Hayward, CA). Intestinal tissue for RNA specimens from patients diagnosed with Crohn's disease and ulcerative colitis was obtained from the National Disease Research Interchange (NDRI) (Philadelphia, PA).

  Astrocytes, lung fibroblasts, skin fibroblasts, coronary artery smooth muscle cells, small airway epithelium, bronchial epithelium, microvascular skin endothelial cells, microvascular lung endothelial cells, human pulmonary artery endothelial cells, human umbilical vein endothelial cells All were purchased from Clonetics (Walkersville, MD) and grown in culture medium supplied by Clonetics for these cell types. These primary cell types were activated with various cytokines or combinations of cytokines for 6 hours and / or 12-14 hours as indicated. The following cytokines were used: approximately 1-5 ng / ml IL-1 beta, approximately 5-10 ng / ml TNF alpha, approximately 20-50 ng / ml IFN gamma, approximately 5-10 ng / ml IL-4. Approximately 5-10 ng / ml IL-9, approximately 5-10 ng / ml IL-13. Sometimes endothelial cells were starved for various times by culturing in basal medium from Clonetics with 0.1% serum.

Mononuclear cells were prepared from the blood of CuraGen Corporation employees using Ficoll. From these cells, DMEM 5% FCS (Hyclone), 100 μM non-essential amino acids (Gibco / Life Technologies, Rockville, MD), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), and LAK cells were prepared by culturing in 10 mM Hepes (Gibco) and interleukin 2 for 4-6 days. Cells were then treated with 10-20 ng / ml PMA and 1-2 μg / ml ionomycin, 5-10 ng / ml IL-12, 20-50 ng / ml IFN-gamma and 5-10 ng / ml IL-18. Either was activated for 6 hours. In some cases, mononuclear cells are allowed to remain in DMEM 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM with approximately 5 μg / ml of PHA (phytohemagglutinin) or PWM (American pokeweed mitogen) for 4-5 days. The cells were cultured in sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), and 10 mM Hepes (Gibco). Samples were taken at 24, 48 and 72 hours for RNA preparation. MLR (mixed lymphocyte reaction) samples were collected from two donors, mononuclear cells were isolated using Ficoll, and the isolated mononuclear cells were 1: 1 with DMEM 5% FCS (Hyclone). ), 100 μM non-essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol (5.5 × 10 ⁻⁵ M) (Gibco), and approximately 2 × 10 ⁶ cells / ml in 10 mM Hepes (Gibco). Was obtained by mixing at a final concentration of. MLRs were cultured and samples were taken at various time points ranging from 1-7 days for RNA preparation.

Monocytes were isolated from mononuclear cells using CD14 Miltenyi Beads, positive veVS selection column and Vario Magnet according to the manufacturer's instructions. Monocytes were isolated from DMEM 5% fetal calf serum (FCS) (Hyclone, Logen, UT), 100 μM non-essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), And were differentiated into dendritic cells by culturing in 10 mM Hepes (Gibco), 50 ng / ml GMCSF and 5 ng / ml IL-4 for 5-7 days. Monocytes were treated with DMEM 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), 10 mM Hepes (Gibco) and 10% AB human. Macrophages were prepared by culturing in serum or approximately 50 ng / ml MCSF for 5-7 days. Monocytes, macrophages and dendritic cells were stimulated with 100 ng / ml lipopolysaccharide (LPS) for 6 hours and 12-14 hours. Dendritic cells were also stimulated with 10 μg / ml anti-CD40 monoclonal antibody for 6 hours and 12-14 hours.

CD4 lymphocytes, CD8 lymphocytes and NK cells were also isolated from mononuclear cells using CD4, CD8 and CD56 Miltenyi Beads, positive VS selection column and Vario Magnet according to the manufacturer's instructions. CD45RA and CD45RO CD4 lymphocytes were isolated by removing CD8, CD56, CD14 and CD19 cells from mononuclear cells using CD8, CD56, CD14 and CD19 Miltenyi Beads and positive selection. Subsequently, when CD45RO CD4 lymphocytes were isolated using CD45RO beads, the remaining cells were CD45RA CD4 lymphocytes. CD45RA CD4, CD45RO CD4 and CD8 lymphocytes were purified from DMEM 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), and 10 mM. Falcon 6 coated in Hepes (Gibco) at 10 ⁶ cells / ml overnight with 0.5 μg / ml anti-CD28 (Pharmingen) and 3 μg / ml anti-CD3 (OKT3, ATCC) in PBS Plated onto well tissue culture plates. Cells were collected for RNA preparation after 6 and 24 hours. To prepare chronically activated CD8 lymphocytes, we activated isolated CD8 lymphocytes on anti-CD28 and anti-CD3 coated plates for 4 days, then collected the cells, They are in DMEM 5% FCS (Hyclone), 100 μM non-essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), and 10 mM Hepes (Gibco) and IL-2 Was grown on. Proliferated CD8 cells were then reactivated for 4 days with anti-CD3 and anti-CD28 bound to the plates and expanded as described above. RNA was isolated 6 and 24 hours after the second activation and 4 days after the second expansion culture. Isolated NK cells, 4-6 days prior to the preparation of RNA, DMEM5% FCS (Hyclone) , 100μM nonessential amino acids (Gibco), 1mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ^- Cultured in ⁵ M (Gibco), and 10 mM Hepes (Gibco) and IL-2.

To obtain B cells, tonsils were obtained from NDRI. The tonsils were cut with sterile dissected scissors and then passed through a sieve. The tonsil cells were then spun down and DMEM 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco) and 10 mM Hepes (Gibco ) At 10 ⁶ cells / ml. In order to activate the cells we used 5 μg / ml PWM or approximately 10 μg / ml anti-CD40 (Pharmingen) and 5-10 ng / ml IL-4. Cells were harvested at 24, 48 and 72 hours for RNA preparation.

To prepare primary and secondary Th1 / Th2 and Tr1 cells, 6-well Falcon plates were coated overnight with 10 μg / ml anti-CD28 (Pharmingen) and 2 μg / ml OKT3 (ATCC) and then with PBS. Washed twice. Umbilical cord blood CD4 lymphocytes (Poietic Systems, German Town, MD), DMEM 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M ( Gibco) was cultured at 10 ⁵ to 10 ⁶ cells / ml in 10 mM Hepes (Gibco) and IL-2 (4 ng / ml). IL-12 (5 ng / ml) and anti-IL4 (1 μg / ml) were used for Th1, while IL-4 (5 ng / ml) and anti-IFN gamma (1 μg / ml) were used for Th2. ) And 5 ng / ml IL-10 for Tr1. After 4-5 days, activated Th1, Th2 and Tr1 lymphocytes were washed once in DMEM and DMEM 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM sodium pyruvate (Gibco), mercapto The cells were cultured in ethanol 5.5 × 10 ⁻⁵ M (Gibco), 10 mM Hepes (Gibco) and IL-2 (1 ng / ml) for 4 to 7 days. Following this, activated Th1, Th2 and Tr1 lymphocytes were added with anti-CD28 / OKT3 and cytokines as described above, but with anti-CD95L (1 μg / ml) to prevent apoptosis, Restimulated for 5 days. After 4-5 days, Th1, Th2 and Tr1 lymphocytes were washed and then re-cultured with IL-2 for 4-7 days. Activated Th1 and Th2 lymphocytes were maintained in this way for up to 3 cycles. From primary and secondary Th1, Th2 and Tr1, 6 and 24 hours after the second and third activation by anti-CD3 and anti-CD28 mAb bound to the plate, and the second and second in interleukin-2. Four days after the start of the third expansion culture, RNA was prepared.

The following white blood cell lines were obtained from ATCC: Ramos, EOL-1, KU-812. EOL cells were further cultured by culturing for 8 days while changing the medium every 3 days at 5 × 10 ⁵ cells / ml in 0.1 mM dbcAMP and adjusting the cell concentration to 5 × 10 ⁵ cells / ml. Differentiated. To cultivate these cells we have 5% FCS (Hyclone), 100 μM non-essential amino acid (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), 10 mM. Hepes (Gibco) was added and DMEM or RPMI (as recommended by ATCC) was used. RNA was prepared from either dormant cells or cells activated with 10 ng / ml PMA and 1 μg / ml ionomycin for 6 and 14 hours. The keratinocyte cell line CCD106 and airway epithelial tumor line NCI-H292 were also obtained from ATCC. Both were cultured in DMEM 5% FCS (Hyclone), 100 μM non-essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 × 10 ⁻⁵ M (Gibco), and 10 mM Hepes (Gibco). . CCD 1106 cells were activated with approximately 5 ng / ml TNF alpha and 1 ng / ml IL-1 beta for 6 and 14 hours, while NCI-H292 cells were activated with the following cytokines for 6 and 14 hours: 5 ng / ml IL- 4, 5 ng / ml IL-9, 5 ng / ml IL-13 and 27 ng / ml IFN gamma.

These cell lines and blood cells, RNA was prepared by dissolving approximately 10 ⁷ cells / ml using Trizol (Gibco BRL). Briefly, 1/10 volume of bromochloropropane (Molecular Research Corporation) was added to the RNA sample, vortexed, and after 10 minutes at room temperature, the tube was centrifuged at 14,000 rpm in a Sorvall SS34 rotator. The aqueous phase was removed and placed in a 15 ml Falcon Tube. An equal volume of isopropanol was added and left at −20 ° C. overnight. The precipitated RNA was spun down in a Sorvall SS34 rotator at 9,000 rpm for 15 minutes and washed in 70% ethanol. The pellet was redissolved in 300 μl RNase-free water and 35 μl buffer (Promega), 5 μl DTT, 7 μl RNAsin and 8 μl DNase were added. Tubes were incubated at 37 ° C. for 30 minutes to remove contaminating genomic DNA, extracted once with phenol / chloroform, and reprecipitated with 1/10 volume of 3M sodium acetate and 2 volumes of 100% ethanol. RNA was spun down and placed in RNase free water. RNA was stored at -80 ° C.

AI_Comprehensive_Panel_v1.0
The plate for AI_Comprehensive_Panel_v1.0 consists of two control wells and 89 copies of cDNA isolated from surgical and autopsy human tissues obtained from Backus Hospital and Clinomics (Frederick, MD) Includes test sample. Total RNA was extracted from tissue samples from Backus Hospital at the CuraGen facility. Total RNA from other tissues was obtained from Clinomics.

  Joint tissue, including synovial fluid, synovium, bone and cartilage, was obtained from a patient who had a total knee or hip replacement operation at Backus Hospital. Immediately thereafter, tissue samples were quickly frozen in liquid nitrogen to ensure that the isolated RNA was of optimal quality and not degraded. Yet another sample of joint tissue from osteoarthritis and rheumatoid arthritis was obtained from Clinomics. Normal control tissues were sourced from Clinomics and were obtained during autopsy of trauma victims.

  Surgical specimens of psoriatic tissue and adjacent matched tissue were obtained from Clinomics as total RNA. Two male and two female patients were selected from age 25 to 47 years. None of the patients were taking prescription drugs when the samples were isolated.

  Surgical specimens of diseased colon and adjacent matched tissue from patients with ulcerative colitis and Crohn's disease were obtained from Clinomics. Intestinal tissue from three women between the ages of 41 and 69 and three male Crohn's disease patients was used. Two patients were not taking prescription drugs, while others were taking dexamethasone, phenobarbital or tylenol. Ulcerative colitis tissue was from 3 male and 4 female patients. Four of the patients took levbid and two received phenobarbital.

  Total RNA from autopsy lung tissue of trauma victims with no disease or emphysema, asthma or COPD was purchased from Clinomics. All emphysema patients ranging from age 40 to 70 years are smokers, this age range focuses on patients with tobacco-related emphysema and excludes those with alpha-1 anti-trypsin deficiency Selected to do. Asthmatic patients ranged from 36 to 75 years of age and excluded smokers to prevent those patients who could have COPD. COPD patients ranged from 35 to 80 years of age and included both smokers and non-smokers. Most patients were taking corticosteroids and bronchodilators.

AI_Comprehensive_Panel_V1.0 The following abbreviations are used in the labels used to identify tissues in the panel.
AI: Autoimmunity Syn: Synovial fluid
Normal: No obvious disease Rep22 / Rep20: Individual patient RA: Rheumatoid arthritis
Backus: From Backus Hospital OA: Osteoarthritis
(SS) (BA) (MF): Individual patient Adj: Adjacent tissue
Match control: Adjacent tissue -M: Male -F: Female COPD: Chronic obstructive pulmonary disease

Panel 5D and 5I
Plates for panels 5D and 5I contain two control wells and various cDNAs isolated from human tissues and cell lines highlighting metabolic disease. Metabolic tissue was obtained from patients enrolled in the Gestational Diabetes study. Cells were obtained during various stages of adipocyte differentiation from human mesenchymal stem cells. Human pancreatic islet cells were also obtained.

In the Gestational Diabetes study, the subjects are young (18-40 years) healthy women with or without gestational diabetes with routine (selective) cesarean section. After childbirth, when the surgical incision is being repaired / sutured, the obstetrician removed a small sample (<1 cc) of metabolic tissue exposed during each surgical level of suturing. The biopsy material was rinsed with sterile saline, blotted and quickly frozen within 5 minutes of extraction. The tissue was then snap frozen in liquid nitrogen, stored individually in sterile screw-cap tubes, and placed on dry ice for transport to CuraGen or until harvested. Metabolic tissues of interest include uterine wall (smooth muscle), visceral fat, skeletal muscle (straight muscle) and subcutaneous fat. The patient's explanation is as follows.
Patient 2 Diabetes Latin American, overweight, not taking insulin Patient 7-9 Non-diabetic Caucasian and obese (BMI> 30)
Patient 10 Diabetes Latin American, overweight, taking insulin Patient 11 Non-diabetic African American overweight Patient 12 Diabetes Latin American taking insulin

Adipocyte differentiation was induced in donor progenitor cells obtained in 3 parts from Osirus (a division of Clonetics / Bio Whittaker), with the exception of donor 3U, which has only two replicas. Clonetics scientists have published human mesenchymal stem cells (HuMSC) for CuraGen, as seen in Mark F. Pittenger, et al., Multilineage Potential of Adult Human Mesenchymal Stem Cells Science Apr 2 1999: 143-147 Isolated, grown and differentiated based on protocol. From Clonetics, Trizol lysates or frozen pellets suitable for mRNA isolation and ds cDNA production were obtained. A general description of each donor is as follows.
Donor 2 and 3U: Mesenchymal stem cells, undifferentiated fat Donor 2 and 3AM: Fat, prematurely differentiated fat Donor 2 and 3AD: Fat, differentiated fat

  Human cell lines are generally obtained from ATCC (American Type Culture Collection), NCI or German cancer cell banks and fall into the following tissue groups: proximal convoluted tubules, uterine smooth muscle cells, small intestine, liver HepG2 Cancer cells, cardiac primary stromal cells and adrenocortical adenoma cells. All of these cells were cultured under standard recommended conditions and RNA was extracted using standard procedures. All samples were processed with CuraGen to produce single stranded cDNA.

  Panel 51 includes the addition of islets from a 58 year old female patient obtained from the Diabetic Research Institute at the University of Miami School of Medicine for all the samples described above. Islet tissue was processed into total RNA by an external vendor and transferred to CuraGen for addition to panel 51.

The following abbreviations are used in labels used to identify tissue in 5D and 5I panels:
GO Adipose: Large retinal fat SK: Skeletal muscle UT: Uterus PL: Placenta AD: Differentiated fat AM: Mid-differentiated fat U: Undifferentiated stem cell

Panel CNSD.01
The plate for panel CNSD.01 contains 94 test samples consisting of 2 control wells and cDNA isolated from autopsy human brain tissue obtained from the Harvard Brain Tissue Resource Center. The brain was removed from the donor calvaria between 4 and 24 hours post-mortem, sectioned by a neuroanatomist and frozen at −80 ° C. in liquid nitrogen vapor. A neuropathologist slices and examines all brains to confirm the diagnosis with a clear associated neuropathology.

  Disease diagnosis is taken from patient records. The panel includes two brains from each of the following diagnoses: Alzheimer's disease, Parkinson's disease, Huntington's disease, progressive supranuclear palsy, depression, and “normal controls”. Within each of these brains, the following areas are presented: zonal gyrus, temporal pole, pallidum, substantia nigra, Brodman 4 area (primary motor area), Brodman 7 area (parietal cortex), Brodman 9 area (frontal frontal) Cortex) and Brodman 17 area (occipital cortex). In all cases, not all of the brain area is presented; for example, Huntington's disease is partly characterized by neurodegeneration in the pallidal sphere, and thus this area cannot be obtained from a confirmed case of Huntington's disease Impossible. Parkinson's disease is also characterized by substantia nigra degeneration, making it more difficult to obtain this area. A neuropathological examination of the normal control brain was performed and found no pathology consistent with neurodegeneration.

The following abbreviations are used in the labels used to identify the tissue in the CNS panel.
PSP: Progressive supranuclear palsy
Sub Nigra: Black quality
Glob Palladus
Temp Pole: Temporal pole
Cing Gyr: Zonal gyrus BA4: Brodman 4 field

Panel CNS_Neurodegeneration_V1.0
Plate for panel CNS_Neurodegenesis_V1.0 is obtained from two control wells and the Harvard Brain Tissue Resource Center (McLean Hospital) and the Human Brain and Spinal Fluid Resource Center (VA Greater Los Angeles Healthcare System) 47 test samples consisting of cDNA isolated from autopsy human brain tissue. The brain was removed from the donor calvaria between 4 and 24 hours post-mortem, sectioned by a neuroanatomist and frozen at -80 ° C in liquid nitrogen vapor. A neuropathologist slices all brains and examines them to confirm the diagnosis with a clear associated neuropathology.

  Disease diagnosis is taken from patient records. The panel includes 6 brains from Alzheimer's disease (AD) disease and 8 brains from “normal controls” that showed no evidence of dementia before death. The eight normal control brains are divided into two categories: controls with no dementia and Alzheimer-like pathology (control) and those with no dementia but severe Alzheimer-like pathology (especially level 3 on a 0-3 scale) Control with evidence of senile plaque load, 0 = no plaque marks, 3 = severe AD senile plaque load). Within each of these brains, the following areas are presented: hippocampus, temporal cortex (Brodman 21), parietal cortex (Brodman 7) and occipital cortex (Brodman 17). These regions were selected to encompass all levels in AD. The hippocampus is an area of early and severe nerve loss in AD; the temporal cortex is known to exhibit neurodegeneration in AD after the hippocampus; the parietal cortex exhibits moderate neuronal death at a late stage of the disease; The occipital cortex survives in AD and therefore serves as a “control” area within AD patients. In all cases, not all brain areas are presented.

The following abbreviations are used in the labels used to identify tissue in the CNS_Neurodegenerative_V1.0 panel.
AD: Alzheimer's disease brain; patient became demented and presented with AD-like pathology at necropsy.
Control: Control brain; non-demented patient, no neuropathology
Control (Path): Control brain; patients with severe AD-like pathology but not dementia
Sup Temporal Ctx: superior temporal cortex
Inf Temporal Ctx: Inferior temporal cortex

A. CG100446-1: Allantocase The expression of the gene CG100446-1 was evaluated using the primer-probe set Ag4178 and is listed in Table AA. The results of RTQ-PCR execution are shown in Table AB.

Continuation of Table AB

  CNS_Neurodegeneration_v1.0 Summary: Ag4178 CG100446-1 gene expression is low / undetectable throughout this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4178 CG10046-01 gene is detected in CNS cancer SK-N-AS cell line (CT = 31.8). In addition, low expression of this gene is also seen in lung cancer cell lines. Therefore, the expression of this gene can be used as a diagnostic marker to detect lung and CNS cancer. Furthermore, therapeutic modulation of this gene may be beneficial in the treatment of lung and brain tumors.

  Moderate expression of this gene is also seen in the testicles. Thus, therapeutic modulation of this gene product may be useful in the treatment of testicular-related diseases such as fertility and hypofunction.

  Panel 4.1D Summary: Ag4178 CG100446-1 gene expression is low / undetectable across all of the samples in this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Ag4178 CG100446-1 gene expression is low / undetectable across all the samples in this panel (CT> 35).

B. CG101025-01 and CG101025-04: calcium / calmodulin-dependent protein kinase type II β chain The expression of the genes CG101025-01 and CG101025-04 was evaluated using the primer-probe set Ag4967 described in Table BA. The results of RTQ-PCR run are shown in Tables BB, BC and BD.

Continuation of Table BB

Continuation of Table BC

General_Screening_Panel_v1.5 Summary: The highest expression of Ag4967 CG101025-01 gene is detected in fetal brain (Ct = 27). High to moderate expression of this gene is found in all areas of the central nervous system tested, including tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. This gene encodes a splice variant of calcium / calmodulin-dependent protein kinase II. Calmodulin (CaM) is the major Ca ^{2+ -binding} protein in the brain and plays an important role in the neural response to changes in Ca ²⁺ concentration in the brain. Calmodulin modulates many Ca ²⁺ -dependent enzymes and is involved in related cellular functions. Among the different CaM-binding proteins, Ca ²⁺ / CaM-dependent protein kinase II and phosphatase calcineurin are particularly important in the brain because of their ubiquitous and involvement in many neural functions (Sola et al ., 2001, Int J Biochem Cell Biol 33 (5): 439-55, PMID: 11331200). Thus, therapeutic modulation of this gene product may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  In addition, moderate expression of this gene is found in heart and skeletal muscle. Thus, the expression of this gene can be used to distinguish brain, heart and skeletal muscle samples from other samples used in this panel. Furthermore, therapeutic regulation of this gene is beneficial for the treatment of heart and muscle related diseases.

  Tumor_cell_strain_screening_panel v3.1 Summary: Ag4967 Expression of this gene is highest in the cerebellum (CT = 28.6), consistent with that observed in panel 1.5. See panel 1.5 for a description of the potential use of this gene in the treatment of central nervous system disorders.

  In addition, the gene is also expressed at moderate levels in 5 out of 13 lung cancer cell lines. Thus, expression of this gene can be used to distinguish lung cancer cell lines from other cell lines on this panel. Furthermore, therapeutic modulation of the activity of this gene or its protein product may be beneficial for the treatment of lung cancer.

  Panel 5 Isle Summary: The highest expression of the Ag4967 CG101025-01 gene is detected in skeletal muscle (CT = 29.7). Moderate expression of this gene is also observed in pancreatic islet cells (Bayer patient 1). Thus, the expression of this gene can be used to distinguish muscle and islet cell samples from the other samples used in this panel.

  This gene encodes a splice variant of calcium / calmodulin-dependent protein kinase II (CaM kinase II). CaM kinase II plays an important role in β-cell insulin exocytosis (Bhatt et al., 2000, Biochem Pharmacol 60 (11): 1655-63, PMID: 11077048). Inhibition of this enzyme suppresses calcium-dependent insulin secretion (Easom, 1999, Diabetes 48 (4): 675-84, PMID: 10102681). Thus, activation of the CG101025-01 gene can be used as a treatment to increase insulin secretion in type 2 diabetes. In addition, therapeutic regulation of this gene is useful for the treatment of muscle related diseases and other endocrine / metabolic related diseases such as obesity.

C. CG101149-01: Calcium Binding Transporter The expression of the gene CG101149-01 was evaluated using the primer-probe set Ag4204 described in Table CA. The results of RTQ-PCR execution are shown in Table CB.

Continuation of Table CB

  CNS_Neurodegeneration_v1.0 Summary: Ag4204 CG101149-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4204 CG101149-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: The highest expression of the Ag4204 CG101149-01 gene is detected in dendritic cells (CT = 28). In addition, moderate to low expression of this gene is found in activated dendritic cells, resting monocytes and macrophages, resting and PMA / ionomycin treated LAK cells, IFNγ and IL-11 treated HUVEC cells, HPAEC cells, 2 It is also found in eosinophils and normal tissues presented by the species WayMLR, PBMC, colon, lung and thymus. Therefore, therapeutic modulation of the gene product through the use of small molecule drugs or antibodies is beneficial for these treatments, and therefore therapeutic agents designed with the protein encoded by this transcript are asthma, emphysema May be important in the treatment of inflammatory bowel disease, arthritis and psoriasis.

  In addition, low level expression of this gene is also observed in cirrhosis (CT = 34.2), but not in normal liver (Panel 1.4D, no expression detected in normal liver). Therefore, therapeutics designed with the protein encoded by this gene can potentially modulate liver function and can be used to identify and treat inflammatory or autoimmune diseases that affect the liver, such as cirrhosis and fibrosis. Have a role.

  General Tumor Screening Panel_v2.4 Summary: Ag4204 CG101149-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

D. CG101169-01: Lipid-binding serum glycoprotein The expression of the gene CG101169-01 was evaluated using the primer-probe set Ag4205 described in Table DA. The results of RTQ-PCR are shown in Table DB.

Continuation of table DB

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4205 CG101169-01 gene is low / undetectable across all samples on this panel (CT> 35).

General_Screening_Panel_v1.4 Overview: Ag4205 CG101169-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: The highest expression of the Ag4205 CG101169-01 gene is detected in the kidney (CT = 30.2). This level of gene expression is also detected in the thymus. Thus, the expression of this gene can be used to distinguish kidney and thymus samples from other samples used in this panel. In addition, therapeutic modulation of the gene product can be used to treat autoimmune and inflammatory diseases that affect the kidney.

  General Tumor Screening Panel_v2.4 Summary: Ag4205 CG101169-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

E. CG101221-01: Novel oxidoreductase UCPA
The expression of the gene CG101221-01 was evaluated using the primer-probe set Ag4219 described in Table EA.

  CNS_Neurodegenesis_v1.0 Summary: Expression of the Ag4219 CG101221-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Overview: Ag4219 CG101221-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag4219 CG101221-01 gene expression is low / undetectable across all of the samples on this panel (CT> 35).

  Panel 5 Islet Summary: Expression of the Ag4219 CG101221-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Ag4219 CG101221-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

F. CG101330-01: Methionyl TRNA formyltransferase The expression of the gene CG101330-01 was evaluated using the primer-probe set Ag4209 described in Table FA. The results of RTQ-PCR execution are shown in Tables FB, FC, FD and FE.

Continuation of Table FC

Continuation of Table FD

  CNS_Neurodegeneration_v1.0 Summary: Ag4209 This panel confirms that the CG101330-01 gene is expressed at low levels in the brain of an independent population. However, no differential expression of this gene was detected between the postmortem Alzheimer's disease brain and the non-demented brain in this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4209 CG101330-01 gene is detected in colon cancer HCT-116 cell line (28.4). Medium level expression of this gene is found in a group of cancer cells derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Therefore, the expression of this gene could be used on this panel to distinguish these samples from other samples and as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at a medium level in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4209 CG101330-01 gene is detected in the INFγ-treated NCI-H292 cell line (CT = 32.2). The gene is expressed at moderate to low levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Is a normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

  General Tumor Screening Panel_v2.4 Summary: The highest expression of Ag4209 CG101330-01 gene is detected in malignant colorectal cancer samples (CT = 33.9). Significant expression of this gene is found in many cancer samples from the kidney, colon, metastatic melanoma and kidney. Therefore, therapeutic modulation of this gene product may be beneficial in the treatment of these cancers.

G. CG101386-01: ABC-2 transporter The expression of the gene CG101386-01 was evaluated using the primer-probe set Ag4210 described in Table GA. The results of RTQ-PCR performed are shown in Tables GB, GC, GD and GE.

Continuation of Table GC

Continuation of Table GD

  CNS_Neurodegeneration_v1.0 Summary: Ag4210 This panel confirms that the CG101386-01 gene is expressed at low levels in the brain of an independent population. However, no differential expression of this gene was detected between the postmortem Alzheimer's disease brain and the non-demented brain in this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4210 CG101386-01 gene is detected in the cerebellum (CT = 28.2). Moderate expression of this gene is found primarily in all areas of the central nervous system such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, the expression of this gene can be used to distinguish brain samples from other samples used in this panel. Furthermore, therapeutic modulation of this gene may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  In addition, moderate to low expression of this gene is found in many cancer cell lines derived from brain, lung, breast, ovarian cancer and melanoma. Thus, expression of this gene can be used as a diagnostic marker for detection of these cancers, and therapeutic modulation of this gene can be beneficial in the treatment of these cancers.

  Panel 4.1D Summary: The highest expression of the Ag4210 CG101386-01 gene is detected in anti-CD40 treated dendritic cells and to a lesser extent in non-activated dendritic cells. This gene is also expressed at moderate levels in LPS activated monocytes but not in resting monocytes. Dendritic cells and activated monocytes play an important role in linking innate immunity to adaptive immunity, but the high expression of this gene in these cells indicates that this gene is immune interventional diseases such as autoimmune diseases and other Suggesting that it may have a role in other T cell mediated diseases such as psoriasis, inflammatory bowel disease and / or bacterial or viral infections. Therefore, modulating the activity of the protein encoded by CG101386-01 with small molecule drugs may be beneficial for the prevention or treatment of these diseases. In addition, the gene is expressed at moderate to low levels in LAK cells, lung fibroblasts, Ramos B cells, macrophages, and normal tissues presented by the colon, lung and kidney. Therefore, the expression of this gene can be used to distinguish these samples from the other samples used in this panel.

  General Tumor Screening Panel_v2.4 Summary: Medium level expression of the Ag4210 CG101386-01 gene is detected only in kidney cancer samples (CT = 32.8). This expression is higher in cancer samples (CT = 40) compared to samples from adjacent normal control tissues. Thus, the expression of this gene can be used to distinguish cancer samples from the control kidney tissue and other samples used in this panel and as a marker for detection of kidney cancer. In addition, therapeutic modulation of this gene may be useful in the treatment of kidney cancer.

H. CG101396-01: Glutamate receptor δ-1
The expression of the gene CG101396-01 was evaluated using the primer-probe set Ag4211 described in Table HA. The results of RTQ-PCR performed are shown in Tables HB, HC, HD, HE and HF.

Continuation of Table HC

Continuation of Table HD

Continuation of Table HE

  CNS_Neurodegenesis_v1.0 Summary: Ag4211 This panel shows no differential expression of the CG101396-01 gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4211 CG101396-01 is found in the fetal brain (CT = 29). In addition, the gene is expressed at moderate levels in all regions of the CNS tested. This gene encodes a protein homologous to the δ2 glutamate receptor and is expressed in the cerebellum. This receptor is involved in behavioral learning and coordination, and synaptic plasticity. Based on the prominent expression of this gene product in the CNS, therapeutic regulation of the expression or function of this gene product is associated with CNS disorders including memory deficits such as Alzheimer's disease and aging, and motor impairment and learning after stroke-induced brain injury. Can be useful to.

  Among tissues with metabolic functions, this gene is expressed at moderate to low levels in the pituitary gland, adipocytes, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscles and heart. Extensive expression in these tissues indicates that this gene product has a role in normal neuroendocrine and metabolic functions, and that unregulated expression of this gene is associated with neuroendocrine disorders or metabolic diseases such as obesity and diabetes. It suggests that it can contribute.

  In addition, moderate levels of expression are seen in a group of samples from ovarian, colon, melanoma and lung cancer cell lines. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of ovarian, colon, melanoma and lung cancer.

  This gene is expressed at a much higher level (CT = 30) in fetal lung tissue compared to expression in adult lung tissue (CT = 35.5). Thus, the expression of this gene can be used to distinguish whether this tissue is of fetal or adult origin.

  Panel 4.1D Summary: Ag4211 The highest expression of this gene is found in the kidney (CT = 30.8). Medium levels of expression are also found in lung and untreated lung microvascular endothelial cells. Low but significant levels of expression are found in untreated and treated astrocytes and Ramos B cells and activated pulmonary microvascular endothelial cells. Astrocyte expression is consistent with the prominent CNS expression seen in panel 1.4. This expression suggests that this gene product may be involved in lung and kidney homeostasis. This therapeutic regulation of protein expression can restore or maintain the function of these organs during inflammation.

  Panel CNS_1 Summary: Ag4211 This panel confirms that this gene is present in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General Tumor Screening Panel_v2.4 Summary: Expression of Ag4211 CG101396-01 gene is found in kidney cancer samples (CT = 31). In addition, moderate to low expression of this gene is detected in melanoma and prostate cancer. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene product may be effective in the treatment of melanoma, kidney and lung cancer and the like.

I. CG101543-01: scop
The expression of the gene CG101543-01 was evaluated using the primer-probe set Ag4216 described in Table IA. The results of RTQ-PCR runs are shown in Tables IB, IC, ID, IE and IF.

Continuation of Table IC

Continuation of table ID

Continuation of Table IE

  CNS_Neurodegeneration_v1.0 Summary: Ag4216 This panel confirms that the CG101543-01 gene is expressed at low levels in the brain of an independent population. This gene is found slightly up-regulated in the temporal cortex of Alzheimer patients. Therefore, neuronal cell death can be reduced by therapeutic regulation of the expression or function of this gene, which is useful for the treatment of this disease.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4216 CG101543-01 gene is detected in thalamus and whole brain (CT = 28). High expression of this gene is found in all areas of the central nervous system tested, including tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. The CG101543-01 gene encodes a homologue of rat SCN circadian oscillatory protein (SCOP). In rats, SCOP has been shown to be predominantly expressed in the brain and is implicated in intracellular signaling in SCN (Shimizu et al., 1999, FEBS Lett 458 (3): 363-9, PMID: 10570941). Thus, therapeutic modulation of the SCOP encoded by this gene may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Medium level expression of this gene is also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at intermediate levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Panel 4.1D Summary: The highest expression of the Ag4216 CG101543-01 gene is detected in activated secondary Th2 cells (CT = 28.8). This gene is expressed at high to moderate levels in a wide range of meaningful cell types in immune responses in healthy and diseased states. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. It is a normal tissue presented. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

  Panel CNS_1 Summary: Ag4216 This panel confirms that the CG101543-01 gene is expressed at low levels in the brain of an independent population. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General Tumor Screening Panel_v2.4 Summary: The highest expression of the Ag4216 CG101543-01 gene is detected in squamous cell carcinoma (CT = 28.9). Interestingly, this expression is higher in cancer samples compared to adjacent control lung tissue (CT = 34.4). Therefore, the expression of this gene can be used for discrimination between these two samples and as a marker for detecting lung cancer.

  Moderate expression of this gene is detected in many normal and cancer samples tested including lung, kidney, colon, prostate and melanoma. Therefore, therapeutic modulation of this gene product may be beneficial in the treatment of these cancers.

J. et al. CG10157-01: Phosphoglycerate Mutase The expression of the gene CG101574-1 was evaluated using the primer-probe set Ag4217 described in Table JA. The results of RTQ-PCR execution are shown in Tables JB, JC and JD.

Continuation of Table JB

Continuation of Table JC

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4217 CG101574-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4217 CG10157-01 gene is detected in gastric cancer NCI-N87 cell line (CT = 29.7). Medium to low level expression of this gene is also found in a group of cancer cells from pancreatic, stomach, colon, lung, kidney, breast, ovarian, melanoma and brain cancers. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at medium to low levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, this gene is expressed at intermediate levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  General_Screening_Panel_v1.5 Summary: Ag4217 CG101574-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: Medium level expression of the Ag4217 CG101574-01 gene is detected only in the kidney (CT = 31.3). Therefore, expression of this gene can be used to distinguish kidney samples from other samples used in this panel. In addition, therapeutic modulation of the gene product may be beneficial for the treatment of autoimmune and inflammatory diseases that affect the kidney, such as lupus and glomerulonephritis.

Panel 5 Islet Summary: Expression of the Ag4217 CG10157-01 gene is low / undetectable across all samples on this panel (CT> 35).
General Tumor Screening Panel_v2.4 Summary: Ag4217 CG10157-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

K. CG101596-01: Lysophosphatidic acid acyltransferase-1
The expression of the gene CG101596-01 was evaluated using the primer-probe set Ag6753 described in Table FA.

  CNS_Neurodegeneration_v1.0 Summary: The results of a single experiment with the Ag6753 CG101596-01 gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

  General_Screening_Panel_v1.6 Summary: The results of a single experiment with the Ag6753 CG101596-01 gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

L. CG101673-1: Bispecific protein phosphatase The expression of the gene CG101673-01 was evaluated using the primer-probe set Ag4222 described in Table LA. The results of the RTQ-PCR run are shown in Tables LB and LC.

Continuation of Table LB

Table LC continued

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4222 CG101673-1 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Low expression of the Ag4222 CG101673-1 gene is found only in the gastric cancer NCI-N87 cell line (CT = 34.7). Therefore, the expression of this gene can be used to distinguish this sample from other samples used in this panel and as a marker for gastric cancer detection. Furthermore, therapeutic modulation of this gene may be useful in the treatment of gastric cancer.

  Panel 4.1D Summary: Low expression of the Ag4222 CG101673-1 gene is found only in the kidney (CT = 30.7). Thus, the expression of this gene can be used to distinguish this sample from the other samples used in this panel. Furthermore, therapeutic modulation of the gene product may be useful for the treatment of autoimmune and inflammatory diseases that affect the kidney, such as lupus and glomerulonephritis.

  General Tumor Screening Panel_v2.4 Summary: Expression of Ag4222 CG101673-1 gene is low / undetectable across all samples on this panel (CT> 35).

M.M. CG101826-01: Adenylate kinase isozyme The expression of the gene CG101826-01 was evaluated using the primer-probe set Ag4226 described in Table MA. The results of RTQ-PCR execution are shown in Tables MB, MC, MD, ME and MF.

Continuation of Table MC

Continuation of table MD

  CNS_Neurodegenesis_v1.0 Summary: Ag4226 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: Ag4226 The highest expression of this gene is found in the thalamus (CT = 25.1), and its expression level is high to moderate in all regions of the CNS tested. This gene encodes a putative adenylate kinase 5, which is a brain-specific enzyme involved in the synthesis of adenine nucleotides required for cellular energy metabolism homeostasis (Van Rompay AR, Eur J Biochem 1999 Apr; 261 (2): 509-17). Inouye S et al., J Neurochem 1998 Jul; 71 (1): 125-33) suggests that this enzyme plays an important role in neuronal function and is involved in neuronal maturation and regeneration. (Inouye S. Biochem Biophys Res Commun 1999 Jan 27; 254 (3): 618-22). Based on its prominent expression in the CNS and its homology to adenylate kinase 5, the expression of this gene could be used to differentiate between neural and non-neural tissues. Furthermore, therapeutic modulation of the gene product expression or function may be useful for the treatment of neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Among tissues with metabolic functions, this gene is expressed at low but significant levels in the pituitary gland, adipocytes, adrenal gland, pancreas, fetal liver, and adult and fetal skeletal muscle and heart. Widespread expression in these tissues means that this gene product can have a role in normal neuroendocrine and metabolic function, and uncontrolled expression of this gene is a neuroendocrine disorder or metabolic disease such as obesity and diabetes This suggests that it can contribute to

  In addition, high levels of expression are seen in a group of samples from brain, kidney and melanoma cancer cell lines. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of brain, kidney and melanoma cancer.

  Panel 4.1D Summary: Ag4226 Expression of this gene is essentially restricted to lung and dermal fibroblasts, with the highest expression found in IL-4 treated dermal fibroblasts (CT = 27.4). The expression in these cells suggests that this gene product may be involved in lung and skin inflammatory conditions. Therefore, the expression of this gene could be used as a marker for fibroblasts. Furthermore, therapeutic modulation of the expression or function of the gene product may be useful in alleviating or eliminating symptoms in patients with asthma, allergies, emphysema and psoriasis.

  Panel 5 Islet Summary: Ag4226 The highest expression of this gene is found in the kidney (CT = 27.4). A low but significant level of expression is also observed in adipocytes. Thus, the expression of this gene can be used to distinguish kidney samples from other samples on this panel.

  General Tumor Screening Panel_v2.4 Overview: Ag4226 The highest expression of this gene is found in samples of normal colon tissue adjacent to colon cancer (CT = 33.1). In addition, low but significant expression is found in normal kidney and prostate, lung and melanoma cancers.

N. CG10186-02: Adenylate Kinase The expression of the gene CG101866-2 was evaluated using the primer-probe set Ag5337 described in Table NA. The results of performing RTQ-PCR are shown in Tables NB, NC and ND.

Continuation of Table NC

Continuation of Table ND

  CNS_Neurodegeneration_v1.0 Summary: Ag5337 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.5 Summary: Ag5337 CG10186-02-02 appears to be expressed primarily in brain-derived tissues, with the highest expression found in the cerebral cortex (CT = 25.7). In addition, high levels of expression are found in the thalamus, substantia nigra, thalamus and tonsils. Medium levels of expression are found in the hippocampus, spinal cord, fetal brain, and brain cancer cell lines. This gene encodes a putative adenylate kinase 5, which is a brain-specific enzyme involved in the synthesis of adenine nucleotides required for cellular energy metabolism homeostasis (Van Rompay AR, Eur J Biochem 1999 Apr; 261 (2): 509-17). Inouye S et al., J Neurochem 1998 Jul; 71 (1): 125-33) suggests that this enzyme plays an important role in neuronal function and is involved in neuronal maturation and regeneration. (Inouye S. Biochem Biophys Res Commun 1999 Jan 27; 254 (3): 618-22). Based on its prominent expression in the CNS and its homology to adenylate kinase 5, the expression of this gene could be used to differentiate between neural and non-neural tissues. Furthermore, therapeutic modulation of the expression or function of the gene product may be useful in the treatment of neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Among tissues with metabolic functions, this gene is expressed at low but significant levels in the pituitary gland, adipocytes, adrenal gland, pancreas, fetal liver, and adult and fetal skeletal muscle and heart. Widespread expression in these tissues means that this gene product can have a role in normal neuroendocrine and metabolic function, and uncontrolled expression of this gene is a neuroendocrine disorder or metabolic disease such as obesity and diabetes This suggests that it can contribute to

  In addition, high levels of expression are seen in a group of samples from brain, kidney and melanoma cancer cell lines. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of brain, kidney and melanoma cancer.

  Panel 4.1D Summary: Moderate expression of the Ag5337 gene is essentially restricted to lung and skin fibroblasts, with the highest expression found in IL-4-treated skin fibroblasts (CT = 28. 3). Medium to low levels of expression are also found in other cells from the lung, such as treated and untreated HPAEC, and lung and skin microvascular endothelial cells. Expression in these cells suggests that this gene product may be involved in lung and skin inflammatory conditions. Thus, the expression of this gene could be used as a marker for fibroblasts. Furthermore, therapeutic modulation of the expression or function of this gene product may be useful to alleviate or eliminate symptoms in patients with asthma, allergies, emphysema and psoriasis.

O. CG102061-01: Glucose-6-phosphate dehydrogenase The expression of the gene CG102061-01 was evaluated using the primer-probe set Ag4230 described in Table OA. The results of RTQ-PCR are shown in Tables OB, OC, OD and OE.

Continuation of Table OC

Continuation of table OD

  CNS_Neurodegeneration_v1.0 Summary: Ag4230 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: Ag4230 The highest expression of this gene is found in skeletal muscle (CT = 26.8). In addition, the gene is expressed at moderate to low levels in the pituitary gland, adipocytes, adrenal glands, pancreas, thyroid, fetal liver, and adult and fetal skeletal muscle and heart. Widespread expression in these tissues means that this gene product can have a role in normal neuroendocrine and metabolic function, and uncontrolled expression of this gene is a neuroendocrine disorder or metabolic disease such as obesity and diabetes This suggests that it can contribute to

  This gene is also expressed at intermediate levels in CNS such as hippocampus, thalamus, substantia nigra, tonsils, cerebellum and cerebral cortex. Thus, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Significant levels of expression are also found in a group of samples from brain, prostate, melanoma, ovary, breast and lung cancer cell lines. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of ovarian cancer, breast cancer and lung cancer.

  Panel 4.1D Summary: Ag4230 The highest expression of this gene is found in IFNγ-treated HUVEC (CT = 31.2). Low but significant levels of expression are also seen in other samples on this panel, such as a group of dermal fibroblasts, lung fibroblasts, and treated and untreated samples of HUVEC. Thus, this gene may be involved in pulmonary and skin inflammatory processes.

  General Tumor Screening Panel_v2.4 Summary: Ag4230 The highest expression of this gene is observed in melanoma (CT = 30.3) and moderate expression is also detected in prostate cancer. In addition, the expression of this gene is overexpressed in lung cancer compared to that in normal adjacent tissues. Therefore, the expression of this gene could be used as a marker for lung cancer. In addition, therapeutic modulation of the expression or function of this gene may be effective in the treatment of lung, prostate, and melanoma cancers.

P. CG102102-01, CG102102-02 and CG102102-02: Peroxisomal short chain alcohol dehydrogenase The expression of the genes CG102102-01, CG102102-02 and CG102102-04 was evaluated using the primer-probe sets Ag4428 and Ag5930 listed in Tables PA and PB did. Note that CG102102-0 is a splice variant of CG102102-01 and is only recognized by probe Ag4428. It should also be noted that CG102102-04 represents a full-length physical clone of the CG102102-01 gene and validates the gene sequence prediction. The results of RTQ-PCR are shown in Tables PC, PD, PE and PF.

Continuation of table PC

Continuation of table PD

Continuation of table PE

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4428 CG102102-01 gene is detected in the breast cancer T47D cell line (CT = 24.5). High level expression of this gene is also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, stomach, colon, lung, kidney, breast, ovarian, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at intermediate levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  General_Screening_Panel_v1.5 Summary: The highest expression of Ag5930 CG102102-01 gene is detected in gastric cancer KATOIII cell line (CT = 29.5). High level expression of this gene is also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, stomach, colon, lung, kidney, breast, ovarian, prostate, squamous cell carcinoma, melanoma and brain cancer.

  In tissues with metabolic or endocrine functions, this gene is expressed at moderate to low levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, this gene is expressed at moderate to low levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 5 Islet Summary: Ag4428 / Ag5930 Two experiments with two different probe and primer sets agree very well with the highest expression in human islet cells (Bayer patient 1) and skeletal muscle (CT = 29. 8-33). This gene encodes peroxisomal short-chain alcohol dehydrogenase. Thus, expression of this gene in human islet cells suggests that the peroxisome oxidation pathway may be important in β-cell physiology. Therefore, therapeutic modulation of this enzyme, or other enzymes in the same pathway, may be useful to enhance insulin secretion in type 2 diabetes.

  In addition, medium level expression of this gene is also observed in most tissues with metabolic / endocrine functions tested, such as adipocytes, placenta, uterus, skeletal muscle and small intestine. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  General Tumor Screening Panel_v2.4 Summary: The highest expression of the Ag5930 CG102102-01 gene is detected in kidney samples (CT = 30). Significant levels of expression of this gene are also found in both normal and cancer samples such as from colon, lung, melanoma, prostate, and kidney. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of lung, prostate, melanoma and some kidney and colon cancer.

Q. CG102102-03: Peroxisomal short chain alcohol dehydrogenase The expression of the gene CG102102-03 was evaluated using the primer-probe sets Ag4441 and Ag4428 described in Tables QA and QB. The results of performing RTQ-PCR are shown in Tables QC, QD and QE. The CG102102-03 gene is a splice variant of the CG102102-01 gene, and the probe Ag4441 is specific for this splice variant.

Continuation of Table QC

  General_Screening_Panel_v1.4 Summary: Ag4428 / Ag4441 Two experiments with different probe and primer sets agree very well with the highest expression of the CG102102-03 gene in the breast cancer T47D cell line (CT = 24.5-26). High level expression of this gene is also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, stomach, colon, lung, kidney, breast, ovarian, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at intermediate levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 5 Islet Summary: Ag4428 The highest expression of this gene is found in human islet cells (Bayer patient 1) (CT = 29.8). This gene encodes peroxisomal short-chain alcohol dehydrogenase. Thus, expression of this gene in human islet cells suggests that the peroxisome oxidation pathway may be important in β-cell physiology. Therefore, therapeutic modulation of this enzyme, or other enzymes in the same pathway, may be useful to enhance insulin secretion in type 2 diabetes.

  In addition, medium level expression of this gene is also observed in most tissues with metabolic / endocrine functions tested, such as adipocytes, placenta, uterus, skeletal muscle and small intestine. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  General Tumor Screening Panel_v2.4 Summary: The highest expression of Ag4441 CG102102-03 gene is found in kidney samples (CT = 27). Significant levels of expression of this gene are also found in both normal and cancer samples from the colon, lung, melanoma, prostate, and kidney. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of colon, lung, prostate, melanoma and kidney cancer.

R. CG102554-01: A novel ATPase associated with various cellular activities
The expression of gene CG102554-01 was evaluated using the primer-probe set Ag4237 described in Table RA. The results of RTQ-PCR execution are shown in Tables RB, RC, RD and RE.

Continuation of Table RC

Continuation of Table RD

  CNS_Neurodegeneration_v1.0 Summary: Ag4237 This panel confirms that the CG102554-01 gene is expressed at low levels in the brain of an independent population. However, no differential expression of this gene was detected between the postmortem Alzheimer's disease brain and the non-demented brain in this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4237 CG101330-01 gene is detected in the breast cancer T47D cell line (CT = 26.6) (CT = 26.6). Medium to high level expression of this gene is also found in a group of cancer cell lines derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at intermediate levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Overview: Two experiments with the same probe and primer set agree well with the highest expression of the gene in activated secondary Th2 and IL-9 treated NCI-H292 (CT = 27-27. 9). The gene is expressed at high to moderate levels in a wide range of cell types that are meaningful in immune responses in healthy and diseased states. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Is a normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

  The results of one experiment with this gene (experiment 175226756) are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

  General tumor screening panel_v2.4 Summary: The highest expression of Ag4237 CG101330-01 is detected in metastatic melanoma (CT = 26.3). Medium to high levels of expression of this gene are also found in cancer samples from the colon, bladder, lung, melanoma, prostate, and kidney. The expression of this gene is higher in cancer samples compared to adjacent normal control tissues. Therefore, the expression of this gene could be used as a marker for detecting the presence of these cancers. In addition, therapeutic modulation of the expression or function of this gene may be effective in the treatment of colon, lung, prostate, bladder, melanoma and kidney cancer.

S. CG102605-01: A novel ATPase associated with various cellular activities
The expression of gene CG102605-01 was evaluated using primer-probe set Ag4240 described in Table SA. The results of the RTQ-PCR run are shown in Tables SB, SC and SD.

Continuation of Table SC

Continuation of Table SD

  CNS_Neurodegeneration_v1.0 Summary: Ag4240 Two experiments with the same probe and primer set agree well. These panels confirm that the CG102605-01 gene is expressed at low levels in the brain of an independent population. However, no differential expression of this gene was detected between the postmortem Alzheimer's disease brain and the non-demented brain in this experiment. See Panel 1.4 for a discussion of potential uses of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4240 CG102605-01 gene is detected in the colon cancer HCT-116 cell line (CT = 30.4). Medium level expression of this gene is also found in a group of cancer cell lines derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. The expression of this gene appears to be higher in cancer cell lines than in normal tissues. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer.

  In tissues with metabolic or endocrine functions, this gene is expressed at moderate to low levels in the pancreas, adipocytes, skeletal muscle, fetal liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Interestingly, this gene is expressed at a much higher level in the fetus (CT = 33.7) compared to adult liver (CT = 39.7). This observation suggests that the expression of this gene can be used to distinguish between fetal and adult skeletal muscle. In addition, the relatively overexpression of the gene in fetal liver suggests that the gene product enhances liver growth or development in the fetus and can also act as a regenerative capacity in adults. Therefore, therapeutic modulation of the protein encoded by this gene would be useful in the treatment of liver related diseases.

  In addition, this gene is expressed at low levels in the fetal and cerebellum. Therefore, therapeutic modulation of this gene product may be useful for the treatment of central nervous system disorders such as ataxia and autism.

  Panel 4.1D Summary: The highest expression of the Ag4240 CG102605-01 gene is detected in the kidney (CT = 31). The gene is expressed at low to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages, and members of the peripheral blood mononuclear cell family, and normal tissue presented by skin fibroblasts and lungs, thymus and kidneys. Furthermore, the expression of this gene is promoted in activated primary and secondary Th1, Th2 and Tr1 cells. This expression pattern suggests that the gene product may be involved in homeostatic processes of these and other cell types and tissues. Thus, regulation of gene products by functional therapeutics can alter the functions associated with these cell types, including asthma, allergies, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, and osteoarthritis May lead to improvement of symptoms in patients with various autoimmune and inflammatory diseases.

  General Tumor Screening Panel_v2.4 Summary: Does not include the results of one experiment with the Ag4240 CG102605-01 gene. According to the amp plot, there were experimental difficulties in performing this experiment.

T.A. CG102909-02: Calcium / calmodulin-dependent protein kinase type II delta chain Expression of the gene CG102909-02 was evaluated using the primer-probe set Ag4392 described in Table TA. The results of RTQ-PCR execution are shown in Table TB.

Continuation of table TB

General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4392 CG102909-02 gene is detected in the cerebellum (CT = 30.4). Moderate expression of this gene is found in all areas of the central nervous system tested, including tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. This gene encodes a splice variant of calcium / calmodulin-dependent protein kinase II. Calmodulin (CaM) is the major Ca ^{2+ -binding} protein in the brain and plays an important role in the neural response to changes in Ca ²⁺ concentration in the brain. Calmodulin modulates many Ca ²⁺ -dependent enzymes and is involved in related cellular functions. Among the different CaM-binding proteins, Ca ²⁺ / CaM-dependent protein kinase II and phosphatase calcineurin are particularly important in the brain because of their ubiquitous and involvement in many neural functions (Sola et al ., 2001, Int J Biochem Cell Biol 33 (5): 439-55, PMID: 11331200). Thus, therapeutic modulation of this gene product may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

U. CG102920-01: short chain 3-hydroxyacyl-CoA dehydrogenase precursor The expression of the gene CG102920-01 was evaluated using the primer-probe set Ag5918 described in Table UA. The results of RTQ-PCR implementation are shown in Tables UB, UC, UD and UE.

Continuation of Table UC

Continuation of Table UD

  CNS_Neurodegeneration_v1.0 Summary: Ag5918 This panel confirms that the CG102920-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.5 Summary: The highest expression of Ag5918 CG102920-01 gene is detected in the skeletal muscle pool. In addition, the gene is expressed at high to medium levels in other tissues with metabolic or endocrine functions, such as pancreas, adipocytes, adrenal gland, thyroid, pituitary, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  The CG102920-01 gene encodes a splice variant of short L-3-hydroxyacyl-CoA dehydrogenase (SCHAD). This gene deficiency is associated with fatty acid oxidation (FAO) disorders such as Ley-like disease (hypoketotic hypoglycemia, hyperammonemia and fatty liver) and cardiomyopathy (Schuler AM, Wood PA, 2002, ILAR J 43 (2): 57-65, PMID: 11917157). Therefore, therapeutic modulation of this gene product may be beneficial in the treatment of FAO defects.

  Medium level expression of this gene is also found in a group of cancer cell lines derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer.

  In addition, the gene is expressed at intermediate levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression. The brain (SCHAD) has been shown to catalyze the oxidation of 17β-estradiol and dihydroandrosterone and alcohols. This will inactivate sex steroid hormones, weakening the protective effects of estrogens and producing aldehydes in neurons. Therefore, what we propose here is that high concentrations of this enzyme in the brain can be a potential risk factor for Alzheimer's disease (He et al., 1999, J Biol Chem 274 (21): 15014 -9, PMID: 10329704).

  Panel 4.1D Summary: The highest expression of the Ag5918 CG102920-01 gene is detected in the kidney (CT = 29.9). The gene is expressed at low to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells are presented by T cells, B cells, endothelial cells, macrophages, and members of the peripheral blood mononuclear cell family, as well as epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. In addition, the expression of this gene is promoted in activated primary and secondary Th1, Th2 and Tr1 cells, PWM / PHA-L treated cells, and PMA / ionomycin treated eosinophils. Thus, modulation of this gene product by functional therapeutics can alter the functions associated with these cell types, including asthma, allergies, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, and osteoarthritis Can lead to improvement of symptoms in patients with autoimmune and inflammatory diseases such as

  Panel 5 Islet Summary: The highest expression of the Ag5918 CG102920-01 gene is detected in islet cells (Bayer patient 1) (CT = 28.4). In addition, medium to low levels of expression of this gene are also found in samples from skeletal muscle, adipocytes, uterus, placenta, liver, heart, and small intestine. Short chain hydroxyacyl CoA dehydrogenase deficiency in humans has been linked to hyperinsulinism (Clayton PT. J Clin Invest. 2001 Aug; 108 (3): 457-65). Therefore, by increasing the activity of this gene product, it is possible to treat hyperinsulinemia, such as diseases of insulin resistance syndrome and type 2 diabetes. See panel 1.4 for a discussion of the use of this gene.

V. CG103051-01: Human ortholog of rat potassium channel The expression of the gene CG103051-01 was evaluated using the primer-probe set Ag4255 described in Table VA. The results of the RTQ-PCR run are shown in Tables VB, VC and VD.

Continuation of Table VC

Continuation of Table VD

  CNS_Neurodegeneration_v1.0 Summary: Ag4255 This panel confirms that the CG103051-1 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of potential uses of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4255 CG103051-01 gene is detected in the cerebellum (CT = 27.3). In addition, the gene is expressed at moderate to high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression. The CG103051-01 gene encodes a homologue of the rat potassium channel subunit. Mutations in the potassium channel subunit KCNQ2, a protein belonging to the potassium channel family, have been implicated in benign familial neonatal convulsions, a dominant inherited form of generalized epilepsy. Thus, similarly, the potassium channel subunit encoded by this gene may have a role in the pathology of neonatal convulsions, and therapeutic regulation of this gene product may be beneficial in the treatment of this disorder .

  In addition, moderate to low expression of this gene is also found in tissues with metabolic or endocrine functions such as adipocytes, pituitary, skeletal muscle, and heart. Thus, therapeutic modulation of this gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Significant expression of this gene is also detected in many cancer cell lines, such as colon cancer, lung cancer and ovarian cancer cell lines. Thus, therapeutic modulation of this gene may be useful in the treatment of these cancers.

  Panel 4.1D Summary: Medium level expression of Ag4255 CG103051-01 is detected only in the kidney (CT = 30.7). Therefore, expression of this gene can be used to distinguish kidneys from other samples used in this panel. Furthermore, therapeutic regulation of this gene may be beneficial in the treatment of autoimmune and inflammatory diseases that affect the kidney, such as lupus and glomerulonephritis.

  Panel 5 Islet Summary: Ag4255 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

W. CG103061-01: Potassium channel The expression of the gene CG103061-01 was evaluated using the primer-probe set Ag4256 described in Table WA.

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4256 CG103061-01 gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4256 CG103061-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: Expression of the Ag4256 CG103061-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  Panel CNS_1 Summary: Expression of Ag4256 CG103061-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Ag4256 CG103061-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

X. CG103061-03: Potassium channel subunit protein The expression of the gene CG103061-03 was evaluated using the primer-probe set Ag5852 described in Table XA.

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag5852 CG103061-03 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: Ag5852 CG103061-03 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: Expression of the Ag5852 CG103061-03 gene is low / undetectable across all of the samples on this panel (CT> 35).

Y. CG103061-05: Potassium channel subunit protein Expression of the gene CG103061-05 was evaluated using the primer-probe set Ag5854 described in Table YA.

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag5854 CG103061-05 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: Ag5854 CG103061-05 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: Expression of the Ag5854 CG103061-05 gene is low / undetectable across all of the samples on this panel (CT> 35).

Z. CG103215-01: Sodium-phosphate cotransporter IIA / C-terminal associated protein 2
The expression of gene CG103215-01 was evaluated using the primer-probe set Ag4257 described in Table ZA.

  CNS_Neurodegeneration_v1.0 Summary: Ag4257 CG103215-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Overview: Ag4257 103215-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag4257 CG103215-01 gene expression is low / undetectable across all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Ag4257 C103215-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

AA. CG103229-01, CG103229-02, and CG103229-06; splice variant of CG103229-01, exon 12-15 deleted, submitted to DDSMT study by cpena on 10/03/01; clone status = FIS; novelty = update variant; ORF start = 12, ORF stop = 1059, frame = 3; 1083 bp
The expression of genes CG103229-01, CG103229-02, and CG103229-06 was evaluated using the primer-probe sets Ag4258, Ag5424 and Ag5429 described in Tables AAA, AAB and AAC. Note that probe Ag5429 is specific for CG103229-02 and CG103229-06 mutants, whereas probe Ag5424 is specific only for mutant CG103229-06. The results of RTQ-PCR performed are shown in Tables AAD and AAE.

Continuation of Table AAE

CNS_Neurodegeneration_v1.0 Summary: Ag5429 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. This gene encodes a homologue of adenylate kinase and is implicated in neuronal function, differentiation, and regeneration (Inouye S, Biochem Biophys Res Commun 1999 Jan 27; 254 (3): 618-22). Thus, therapeutic modulation of the expression or function of the gene product may be effective in the treatment of neurodegenerative disorders including Alzheimer's disease and Parkinson's disease.
Ag4258 / Ag5425 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4258 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).

General_Screening_Panel_v1.5 Summary: Ag5429 The expression of this gene is highest in breast cancer cell lines (CT = 31.6). Medium levels of expression are also found in cell lines derived from ovarian and lung cancer. Thus, modulation of the expression or function of the gene product may be effective in the treatment of these cancers.
The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

Panel 4.1D Summary: Ag4258 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).
The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).
Ag5429 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

AB. CG103229-03: splice variant of CG103229-01 The expression of the gene CG103229-03 was evaluated using the primer-probe set Ag5427 described in Table ABA.

  CNS_Neurodegeneration_v1.0 Summary: Ag5427 Expression of this gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: Ag5427 The expression of this gene is low / undetectable across all of the samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag5427 The expression of this gene is low / undetectable across all samples on this panel (CT> 35).

AC. CG103229-04: splice variant of CG103229-01 The expression of the gene CG103229-04 was evaluated using the primer-probe sets Ag4258 and Ag5426 described in Tables ACA and ACB.

  CNS_Neurodegeneration_v1.0 Summary: Ag5423 / Ag5426 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4258 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: Ag5423 / Ag5426 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag5423 / Ag5426 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Expression of Ag4258 This gene is low / undetectable in all of the samples on this panel (CT> 35).

AD. CG103229-05: splice variant of CG103229-01 The expression of the gene CG103229-05 was evaluated using the primer-probe sets Ag5423 and Ag5426 described in Tables ADA and ADB.

  CNS_Neurodegeneration_v1.0 Summary: Ag5423 / Ag6426 Expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: Ag5423 / Ag6426 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag5423 / Ag5426 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

AE. CG103229-07: splice variant of CG103229-01 The expression of the gene CG103229-07 was evaluated using the primer-probe sets Ag4258, Ag5428 and Ag5429 described in Tables AEA, AEB and AEC. The results of RTQ-PCR runs are shown in Tables AED and AEE.

Continuation of Table AEE

CNS_Neurodegeneration_v1.0 Summary: Ag5429 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. This gene encodes a homologue of adenylate kinase and is implicated in neuronal function, differentiation, and regeneration (Inouye S, Biochem Biophys Res Commun 1999 Jan 27; 254 (3): 618-22). Thus, therapeutic modulation of the expression or function of the gene product may be effective in the treatment of neurodegenerative disorders including Alzheimer's disease and Parkinson's disease.
Ag4258 / Ag5428 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4258 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).

General_Screening_Panel_v1.5 Summary: Ag5429 The expression of this gene is highest in breast cancer cell lines (CT = 31.6). Medium levels of expression are also found in cell lines derived from ovarian and lung cancer. Thus, modulation of the expression or function of the gene product may be effective in the treatment of these cancers.
Ag5428 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

Panel 4.1D Summary: Ag4258 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).
Ag5428 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).
Ag5429 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Expression of Ag4258 This gene is low / undetectable in all of the samples on this panel (CT> 35).

AF. CG103229-08: CG103229-01 splice variant The expression of the gene CG103229-08 was evaluated using the primer-probe sets Ag4258 and Ag5425 described in Tables AFA and AFB. The results of RTQ-PCR runs are shown in Tables AFC and AFD.

Continuation of Table AFD

CNS_Neurodegeneration_v1.0 Summary: Ag5429 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. This gene encodes a homologue of adenylate kinase and is implicated in neuronal function, differentiation, and regeneration (Inouye S, Biochem Biophys Res Commun 1999 Jan 27; 254 (3): 618-22). Thus, therapeutic modulation of the expression or function of the gene product may be effective in the treatment of neurodegenerative disorders including Alzheimer's disease and Parkinson's disease.
Ag4258 / Ag5425 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4258 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: The highest expression of Ag5425 CG103229-08 is detected in the lung cancer LX-1 cell line (CT = 34.2). Low but significant levels of expression of this gene are also found in some cancer cell lines derived from fetal lung, trachea, and pancreas, lung and ovarian cancer. Thus, therapeutic regulation of this gene may be useful for the treatment of pancreatic, lung and ovarian cancer.

  Interestingly, this gene is expressed at a much higher level (CT = 34.6) in the fetus compared to the adult lung (CT = 40). This observation suggests that this gene can be used to distinguish between adult and fetal lungs. In addition, the relatively overexpression of this gene in the fetal lung suggests that the protein product can enhance fetal lung growth and development and may also affect adult regenerative capacity. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in the treatment of lung related diseases.

Panel 4.1D Summary: Ag4258 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).
Ag5425 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).
Ag5429 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Summary: Expression of Ag4258 This gene is low / undetectable in all of the samples on this panel (CT> 35).

AG. CG103229-09: CG103229-01 splice variant The expression of the gene CG103229-09 was evaluated using the primer-probe set Ag6120 described in Table AGA. The results of RTQ-PCR are shown in Tables AGB and AGC.

Continuation of Table AGC

  CNS_Neurodegeneration_v1.0 Summary: Ag6120 This panel confirms the presence of CG103229-09 variants at low but significant levels in the brain. For a discussion of the use of this putative adenylate kinase in the central nervous system, see CG103229-06.

  General_Screening_Panel_v1.5 Summary: High expression of Ag6120 CG103229-09 gene is found in fetal kidney (CT = 32). Therefore, expression of this gene could be used to distinguish between adult kidney (CT = 35.4) and fetal kidney. Low but significant levels of expression are also found in other samples such as pancreas, adrenal gland, fetal and adult lung and pancreatic cancer, kidney, lung and ovarian cancer cell lines. Expression in multiple cancer cell lines suggests that this gene product may be involved in cell growth and / or proliferation.

  Panel 4.1D Summary: Ag6120 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

AH. CG103285-01, CG103285-02 and CG103285-03: Ubiquitin carboxyl terminal hydrolase Genes CG103285-01, CG103285-02 and CG103285-03 are expressed using the primer-probe sets Ag1472 and Ag963 described in Tables AHA and AHB And evaluated. The results of performing RTQ-PCR are shown in Tables AHC, AHD, AHE and AHF.

Continuation of Table AHD

Continuation of Table AHE

  CNS_Neurodegenesis_v1.0 Summary: Ag 1472 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: Ag1472 This gene is expressed at high to medium levels in all of the samples in this panel. The highest expression is detected in gastric cancer cell lines (CT = 24). This gene is also highly expressed in all cell lines on this panel, with higher expression being observed in cancer cell lines than in normal tissue samples. This ubiquitous expression pattern suggests a role for this protein product in cell survival and proliferation. Therapeutic modulation of this gene or its protein product may be useful for the treatment of cancer.

  Among tissues with metabolic functions, this gene is expressed at high to medium levels in the pituitary gland, adipocytes, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. Widespread expression in these tissues indicates that this gene product has a role in normal neuroendocrine and metabolic functions, and uncontrolled expression of this gene is associated with neuroendocrine disorders or metabolic diseases such as obesity and diabetes. It suggests that it can contribute.

  This gene is also highly expressed in CNS such as hippocampus, thalamus, substantia nigra, tonsils, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful for the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  In addition, this gene is expressed at a much higher level (CT = 25.8) in the fetal lung compared to expression in the adult lung (CT = 28.8). Thus, the expression of this gene can be used to distinguish whether this tissue is of fetal or adult origin.

  Tumor_cell_strain_screening_panel_v3.1 Summary: Ag1472 The highest expression is found in the medulloblastoma cell line (CT = 24.7). Overall, the expression in this panel is ubiquitous and is consistent with the expression in panel 1.4. See this panel for a discussion of this gene's use in cancer.

  General Tumor Screening Panel_v2.4 Summary: Ag1472 The highest expression of this gene is found in lung cancer (CT = 25.3). In addition, this gene is overexpressed in lung and kidney cancers compared to expression in normal adjacent tissues. This gene is also highly expressed in melanoma and prostate cancer. Therefore, expression of this gene could be used as a marker for these cancers. Therapeutic modulation of the expression or function of the gene product may also be effective in the treatment of lung, kidney, prostate, and melanoma cancers.

AI. CG103374-01: Kinesin light chain 3 (KLC3)
The expression of gene CG103374-01 was evaluated using the primer-probe set Ag4259 described in Table AIA.

  CNS_Neurodegeneration_v1.0 Summary: Ag4259 Expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4259 Expression of this gene is low / undetectable in all samples on this panel (CT> 35).

  Panel 4.1D Summary: Ag4259 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General Tumor Screening Panel_v2.4 Overview: Ag4259 Expression of this gene is low / undetectable across all samples on this panel (CT> 35).

AJ. CG103415-01 and CG103415-02: Novel ion transporter proteins The expression of the genes CG103415-01 and CG103415-02 was evaluated using the primer-probe set Ag4260 described in Table AJA. The results of RTQ-PCR execution are shown in Table AJB. Note also that CG103415-02 represents a full-length physical clone of the CG103415-01 gene, validating the prediction of the gene sequence.

Continuation of Table AJB

  CNS_Neurodegeneration_v1.0 Summary: Ag4260 Expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4260 The expression of this gene is low / undetectable in all the samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag4260 Expression of this gene is restricted to memory T cells (CT = 32.9) and IFNγ stimulated skin fibroblasts (CT = 39.4). Thus, the expression of this gene could be used to distinguish these samples from other samples on this panel. In addition, therapeutic modulation of this gene may be useful in the treatment of asthma, emphysema, IBD, lupus or arthritis, psoriasis, and other diseases in which T cells are activated.

  General Tumor Screening Panel_v2.4 Summary: Ag4260 The expression of this gene is low / undetectable across all samples on this panel (CT> 35).

AK. CG103481-01 and CG103481-02: Expression of the novel Meis3-lipocalin genes CG103481-01 and CG103481-01 was evaluated using the primer-probe set Ag4263 described in Table AKA. Note also that CG103481-02 represents a full-length physical clone of the CG103481-01 gene, validating the prediction of the gene sequence.

  CNS_Neurodegeneration_v1.0 Summary: Ag4263 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35). The amplifier plot suggests that there is a high probability of probe failure.

  General_Screening_Panel_v1.4 Summary: Ag4263 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35). The amplifier plot suggests that there is a high probability of probe failure.

  Panel 4.1D Summary: Ag4263 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35). The amplifier plot suggests that there is a high probability of probe failure.

  Panel CNS_1 Summary: Ag4263 The expression of this gene is low / undetectable across all samples on this panel (CT> 35). The amplifier plot suggests that there is a high probability of probe failure.

AL. CG103900-01: FYVE finger-containing phosphoinositide kinase The expression of the gene CG103900-01 was evaluated using the primer-probe set Ag4265 described in Table ALA. The results of the RTQ-PCR run are shown in Tables ALB, ALC and ALD.

Continuation of Table ALC

Continuation of Table ALD

  CNS_Neurodegeneration_v1.0 Summary: Ag4265 This panel confirms that the CG103900-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4265 CG103900-01 gene is detected in CNS cancer (Glio) SF-295 and melanoma SK-MEL-5 cell lines (CT = 26.3) . High level expression of this gene is found in a group of cancer cells derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at high levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes. The CG103900-01 gene encodes a homologue of mouse FYVE finger-containing phosphoinositide kinase (PIKfyve). Mouse PIKfyve was first isolated from a mouse adipocyte cDNA library. It has been found to be abundant in insulin-sensitive cells and tissues and is involved in the insulin-signaling pathway (Shisheva et al., 1999, Mol Cell Biol 19 (1): 623-34, PMID : 9858586; Sbrissa et al, 2001, Mol Cell Endocrinol 181 (1-2): 35-46, MPID: 11476939). Therefore, like the mouse protein, the PIKfyve protein encoded by this gene may have a role in the insulin-signaling pathway.

  Interestingly, this gene is expressed at a much higher level in fetuses (CT = 27-28.6) compared to adult lung and liver (CT = 30-33). This observation suggests that the expression of this gene can be used to distinguish fetal lung and liver from adults. In addition, the relative overexpression of this gene in fetal tissue suggests that the protein product promotes lung and liver growth and development in the fetus and may also affect the regenerative capacity in adults. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in the treatment of liver and lung related diseases.

  In addition, expression of this gene is found at high levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4265 CG103900-01 gene is detected in PMA / ionomycin treated basophils (CT = 27.8). The gene is expressed at high to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Is a normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change functions associated with these cell types, such as asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

AM. CG104220-01: equilibrium nucleoside transporter 3
The expression of gene CG104220-01 was evaluated using the primer-probe set Ag5919 described in Table AMA. The results of RTQ-PCR performed are shown in Tables AMB and AMC.

Continuation of Table AMB

  General_Screening_Panel_v1.5 Summary: The Ag5919 CG104220-01 gene is widely expressed in this panel, with the highest expression found in liver cancer cell lines (CT = 28.8). A medium level of expression is seen in all cell lines on this panel. This gene encodes a putative equilibrium nucleoside transporter that mediates cellular uptake of anticancer nucleoside analogs. Since this gene is expressed in cancer cell lines, therapeutic modulation of the function of this gene product may be useful in the treatment of these cancers.

  Among tissues with metabolic functions, this gene is expressed at low but significant levels in adipocytes, adrenal glands, pancreas, thyroid, and adult and fetal skeletal muscle, heart and liver. Extensive expression in these tissues suggests that this gene product may have a role in normal neuroendocrine and metabolic functions. Parodi et al. Demonstrated that inhibition of this transporter could contribute to the vasodilation seen in diabetes (Circ Res 2002 Mar 22; 90 (5): 570-7). Furthermore, this transporter can be down-regulated in some diabetes (Osses N. Reprod Fertil Dev 1995; 7 (6): 1499-503). Thus, therapeutic modulation of the expression or function of this protein may be effective in the treatment of neuroendocrine disorders or metabolic diseases such as obesity and diabetes.

  In addition, this gene is expressed at a medium to low level in CNS such as hippocampus, thalamus, substantia nigra, tonsils, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful for the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Panel 5 Isle Summary: Ag5919 The highest expression of this gene is found in the hepatocyte cell line (CT = 29.8). Thus, the expression of this gene could be used to distinguish this sample from other samples on this panel. In addition, a low but significant level of expression is observed in adipocytes. See panel 1.5 for a discussion of the use of this gene in metabolic diseases.

AN. CG104220-02: Equilibrium nucleoside transporter 3
The expression of gene CG104220-02 was evaluated using the primer-probe set Ag5931 described in Table ANA. The results of RTQ-PCR implementation are shown in Tables ANB, ANC, AND and ANE.

Continuation of Table ANC

Continuation of table AND

  CNS_Neurodegeneration_v1.0 Summary: Ag5931 This panel does not show differential expression of this gene in Alzheimer's disease. However, this panel shows that this gene is expressed at low levels in the brain. Thus, this gene is useful for distinguishing brain tissue from non-neural tissue and may be useful as a drug target in neurodegenerative diseases.

  General_Screening_Panel_v1.5 Summary: The expression of the Ag5931 CG104220-02 gene is much higher on this panel in brain cancer cell lines than in other samples (CT = 26). Therefore, the expression of this gene could be used to distinguish this sample from other samples on this panel. Furthermore, therapeutic modulation of the expression or function of this gene could be used to treat brain cancer.

  Panel 4.1D Summary: Ag5931 The highest expression of this mutant is found in untreated dendritic cells (CT = 33). Low but significant levels of expression are also observed in resting macrophages, LAK cells, and treated and untreated NCI-H292 cells. The expression of this gene in these lineages of resting cells suggests that the protein encoded by this transcript may be involved in normal immune processes associated with immune homeostasis. This gene encodes a protein involved in macrophage activation and proliferation, a putative equilibrium nucleotide transporter (Soler C. FASEB J 2001 Sep; 15 (11): 1979-88). Therefore, agonist (ligand-like) therapeutics designed with this protein product activate / stimulate the immune response and improve the effectiveness of vaccines and antiviral or antimicrobial treatments. In addition, expression in dendritic cells indicates that the therapeutic use of the protein encoded by this transcript is immunomodulation, organ / bone marrow transplantation, and antigen presentation is a function of mature dendritic cells asthma, rheumatoid arthritis Suggests that it may be important in the treatment of diseases when it plays important roles such as IBD, allergies, and psoriasis.

  Panel 5 Islet Summary: Ag5931 CG104220-02 mutant is detected only in hepatocyte cell lines (CT = 32.8). Thus, the expression of this gene could be used on this panel to distinguish this sample from other samples. In addition, therapeutic modulation of this gene product may be useful in the treatment of liver related diseases.

AO. CG104693-01 and CG104693-02: Adenylate Kinase The expression of the genes CG104693-01 and CG104693-02 was evaluated using the primer-probe sets Ag4271 and Ag5975 described in Tables AOA and AOB. Note that probe Ag5975 is specific for mutant CG10469-02. The results of RTQ-PCR execution are shown in Tables AOC, AOD, AOE and AOF.

Continuation of Table AOC

Continuation of Table AOE

Continuation of Table AOF

  AI_Comprehensive Panel_v1.0 Summary: Expression of the Ag5975 CG10469-03-02 gene is limited to a few samples of normal tissue adjacent to either psoriasis or Crohn's disease (CT = 34.6). Expression in the other samples was low / undetectable. See Panel 4.1D for a discussion of the use of this gene in inflammation.

  CNS_Neurodegenesis_v1.0 Summary: Ag4271 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the low level presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

General_Screening_Panel_v1.4 Summary: Ag4271 The highest expression of this gene is found in breast cancer cell lines (CT = 28) and significant expression is also detected in ovarian cancer cell lines.
Therefore, the expression of this gene could be used on this panel to distinguish breast cancer cell lines from other samples and as a marker for breast cancer. Therapeutic modulation of the expression or function of this gene may be effective in the treatment of breast and ovarian cancer.

  This gene is also expressed at moderate to low levels in CNS such as hippocampus, thalamus, substantia nigra, tonsils, cerebellum and cerebral cortex. This gene encodes a putative adenylate kinase and may be involved in neuronal maturation and regeneration (Inouye S. Biochem Biophys Res Commun 1999 Jan 27; 254 (3): 618-22). Thus, therapeutic modulation of the expression or function of this gene may be useful for the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Among tissues with metabolic functions, this gene is expressed at moderate to low levels in the pituitary gland, adipocytes, adrenal gland, pancreas, thyroid, fetal skeletal muscle, and adult and fetal heart. Expression in these tissues indicates that this gene product may have a role in normal neuroendocrine and metabolic functions, and uncontrolled expression of this gene contributes to neuroendocrine disorders or metabolic diseases such as obesity and diabetes This suggests that it may be.

Panel 4.1D Summary: Ag4271 The highest expression of this gene is found in the KU-812 basophil cell line (CT = 31.2). In addition, low to moderate expression is found in the kidney and thymus. Therefore, the expression of this gene could be used on this panel to distinguish basophil samples from other samples and as a potential marker for these cells.
The expression of Ag5975 CG10469-02 is detected exclusively in the kidney (CT = 34.5). Therefore, this gene may be involved in the homeostasis of this organ.

AP. CG104790-01: vacuolar ATP synthase-like protein The expression of the gene CG104790-01 was evaluated using the primer-probe set Ag4272 described in Table APA. The results of RTQ-PCR are shown in Table APB.

Continuation of table APB

CNS_Neurodegeneration_v1.0 Summary: Ag4272 The expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).
General_Screening_Panel_v1.4 Summary: Ag4272 Expression of this gene is restricted to fetal kidney and colon and liver cancer cell lines (CT = 30-33). Therefore, the expression of this gene could be used on this panel to distinguish these samples from other samples and to distinguish between fetal kidney tissue and adult kidney tissue.
Panel 4.1D Summary: Ag4272 The results of one experiment with this gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

AQ. CG104800-01 and CG104800-02: Polypeptide N-acetylgalactosaminyltransferase The expression of the genes CG104800-01 and CG104800-02 uses the primer-probe sets Ag3484, Ag4911 and Ag4910 described in Tables AQA, AQB and AQC. And evaluated. The results of performing RTQ-PCR are shown in Tables AQD, AQE, AQF, AQG and AQH. Note also that CG104800-02 represents a full-length physical clone of the CG104800-01 gene, validating the prediction of the gene sequence.

Continuation of table AQE

Continuation of table AQF

Continuation of table AQG

Continuation of table AQG

Continuation of Table AQH

  CNS_Neurodegenesis_v1.0 Summary: Ag3484 / Ag4911 Two experiments with two different probe and primer sets give very consistent results. This panel shows no differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag3484 CG104800-01 gene is found in brain cancer cell lines (CT = 28.4). Medium levels of expression are also found in cell lines derived from lung cancer, breast cancer and melanoma. Therefore, the expression of this gene could be used on this panel to distinguish brain cancer samples from other samples and as a marker for brain cancer. In addition, therapeutic modulation of the expression or function of the gene product may be useful in the treatment of these cancers.

  Low but significant levels of expression are detected in the pancreas, adipocytes, adrenal gland and pituitary gland. This expression suggests that the gene product may be involved in the pathogenesis and / or treatment of metabolic disorders such as obesity and diabetes.

  This gene is also expressed at medium levels in all regions of the CNS tested. This gene encodes a putative N-acetylgalactosaminyltransferase that can catalyze O-glycosylation in the brain. Therefore, therapeutic modulation of the expression or function of this gene may be useful for the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  General_Screening_Panel_v1.5 Summary: Ag4910 / Ag4911 Two experiments give very consistent results. The highest expression is found in brain-derived tissue, with the first experiment being the highest in the cerebellum (CT = 27.4) and the second experiment being the highest in the brain cancer cell line (CT = 26). Expression is consistent with that of panel 1.4. See 1.4 for a discussion of the use of this gene in cancer, metabolic diseases and the central nervous system.

  Tumor_Cell_Strain_Screening_Panel_v3.1 Summary: Ag4910 / Ag4911 Two experiments with the same probe and primer give very consistent results. The highest expression of this gene is found in samples from the cerebellum (CT = 27). In addition, a significant level of expression is observed in a group of cell lines derived from lung cancer. See panel 1.4 for a discussion of the use of this gene in cancer.

  Panel 4D Summary: Ag3484 The highest expression of this gene is found in TNF-α and IL-1β stimulated astrocytes (CT = 30.7). Medium levels of expression are also found in the colon. Low but significant levels of expression are found in keratinocytes, NCI-H292 cells, lung, and Crohn's disease. The expression of this gene appears to be up-regulated in TNF-α / IL-1b treated astrocytes compared to that in resting astrocytes. Thus, this transcript can function in astrocyte differentiation and activation. Therefore, therapeutic regulation of this transcript or the design of therapeutic agents with encoded proteins would be important for the treatment of multiple sclerosis or other inflammatory diseases of the CNS.

  General Tumor Screening Panel_v2.4 Summary: Ag3484 One experimental result with this gene is not included. According to the amp plot, there were experimental difficulties in performing this experiment.

AR. CG104813-1: Phosphatidylethanolamine binding protein-like protein The expression of the gene CG104813-1-01 was evaluated using the primer-probe set Ag4277 described in Table ARA. The results of RTQ-PCR runs are shown in Tables ARB, ARC and ARD.

Continuation of Table ARC

  CNS_Neurodegeneration_v1.0 Summary: Ag4277 This panel confirms that the CG104813-1 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4277 CG104813-1-01 gene is detected in the thymus (CT = 29.5). In addition, high expression of this gene is also observed in the spleen (CT = 29.7). Thus, the expression of this gene can be used in this panel to distinguish this sample from other samples. In addition, the expression of this gene in the thymus and spleen suggests that it may play an important role in T and B cell development. Therefore, therapeutic regulation of this gene may be useful for the treatment of allergies, autoimmune diseases, and inflammatory diseases.

  Medium level expression of this gene is also detected in colon cancer, CNS cancer and colon cancer cell lines. Thus, therapeutic modulation of this gene through the use of small molecule drugs may be beneficial for the treatment of colorectal and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at low to medium levels in the pancreas, adipocytes, heart, fetal liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, this gene is expressed at moderate to low levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression. The CG104813-1 gene encodes a homologue of phosphatidylethanolamine binding protein (PEBP) (hippocampal cholinergic neurostimulatory peptide) (HCNP) (RAF kinase inhibitor protein) (RKIP). PEBP is involved in septic hippocampal phenotypic development, and PEBP mRNA down-regulation is associated with the pathogenic process behind Alzheimer's disease (AD) (Maki et al., 2002, J Neuropathol Exp Neurol 61 (2): 176-85; PMID: 11853019). Therefore, based on homology, the PEBP-like protein encoded by this gene has a role in the development of the septic hippocampal system, and therapeutic regulation that increases its expression and function may be useful in the treatment of AD.

  General Tumor Screening Panel_v2.4 Summary: The highest expression of Ag4277 CG104813-1-01 gene is detected in kidney cancer (CT = 30.8). In addition, significant expression of this gene is also found in kidney, lung, colon, melanoma and prostate cancer. The expression of this gene is higher in cancer compared to adjacent normal tissue samples. Therefore, expression of this gene can be used as a diagnostic marker to detect these cancers, and therapeutic regulation of this gene is beneficial for the treatment of kidney, lung, colon, melanoma, and prostate cancer It can be.

AS. CG104892-1 and CG104892-02: haloacid dehalogenase The expression of the genes CG104892-01 and CG104892-02 was evaluated using the primer-probe set Ag4273 described in Table ASA. The results of performing RTQ-PCR are shown in Tables ASB, ASC and ASD. Note also that CG104892-02 represents a full-length physical clone of the CG104892-01 gene, validating the prediction of the gene sequence.

Continuation of Table ASC

Continuation of Table ASD

  CNS_Neurodegeneration_v1.0 Summary: Ag 4273 This panel confirms that the CG104892-1 gene is expressed at low levels in the brain of an independent population. This gene is found to be slightly up-regulated in the temporal cortex of Alzheimer's disease patients. Therefore, therapeutic modulation of the expression or function of this gene reduces neuronal cell death and may be useful in the treatment of this disease.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4273 CG104892-1 gene is detected in the CNS cancer SNB-75 cell line (CT = 27.7). High to moderate expression of this gene is also found in a group of cancer cells derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at a medium level in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Interestingly, this gene is expressed at a much higher level in the fetus (CT = 30) than in the adult liver (CT = 34.7). This observation suggests that expression of this gene can be used to discriminate between fetus and adult liver. In addition, the relatively overexpression of this gene in fetal liver suggests that this protein product enhances fetal liver growth and development and, as a result, can act as a regenerative capacity in adults. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in the treatment of liver related diseases.

  In addition, this gene is expressed at moderate to low levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4273 CG10489-01 gene is detected in activated CD45RO CD4 lymphocytes (CT = 31.3). The gene is expressed at high to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells are presented by T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, as well as epithelial and fibroblast types from lung and skin, and lung, thymus and kidney Normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

AT. CG104955-01: 10-formyltetrahydrofolate dehydrogenase The expression of the gene CG104955-01 was evaluated using the primer-probe set Ag4303 described in Table ATA. The results of the RTQ-PCR run are shown in Tables ATB, ATC and ATD.

Continuation of table ATB

  General_Screening_Panel_v1.4 Summary: Ag4303 The highest expression of this gene is detected in CNS cancer SF-295 cell line (CT = 25). In addition, moderate to high expression of this gene is also found in a group of cancer cells derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at a medium level in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, fetal liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Interestingly, this gene is expressed at a much higher level in the fetus (CT = 34) than in the adult liver (CT = 40). This observation suggests that expression of this gene can be used to discriminate between fetus and adult liver. In addition, the relatively overexpression of this gene in fetal liver suggests that this protein product enhances fetal liver growth and development and, as a result, can act as a regenerative capacity in adults. Therefore, therapeutic modulation of the protein encoded by this gene could be useful in the treatment of liver related diseases.

  In addition, the gene is expressed at intermediate levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 5 Islet Summary: Ag4303 The highest expression of this gene is detected in adipocytes (CT = 28.4). High to moderate expression of this gene is found in skeletal muscle, islet cells, adipocytes, small intestine and mesenchymal stem cells. Therefore, therapeutic modulation of this gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

General Tumor Screening Panel_v2.4 Summary: Ag4303 The highest expression of this gene is detected in prostate cancer (CT = 29.3). For some cancer samples from patients with kidney, lung, melanoma, bladder and prostate cancer, this gene has a higher expression compared to normal adjacent tissues of these patients.
Therefore, the expression of this gene can be used as a cancer diagnostic marker. Furthermore, therapeutic modulation of this gene using small molecules alone or in combination may be beneficial for the treatment of these cancers.

AU. CG1050556-01: 17β-hydroxysteroid dehydrogenase type VII Expression of the gene CG1050656-01 was evaluated using the primer-probe set Ag5926 described in Table AUA. The results of RTQ-PCR execution are shown in Tables AUB, AUC, AUD and AUE.

Continuation of table AUC

Continuation of Table AUD

  CNS_Neurodegenesis_v1.0 Summary: Ag5926 Two experiments with the same probe and primer set agree very well. This gene appears to be upregulated in the temporal cortex of Alzheimer patients. Thus, neuronal cell death can be reduced by therapeutic regulation of the expression or function of this gene and is useful in the treatment of this disease.

  General_Screening_Panel_v1.5 Summary: Ag5926 This gene is widely expressed at low but significant levels in this panel and is most expressed in liver cancer cell lines (CT = 32). Significant levels of expression are also found in breast and colon cancer cell lines. This gene encodes an enzyme involved in estrogen metabolism, a putative 17β hydroxysteroid dehydrogenase. Gunnarsson et al. Suggest that altered expression of this enzyme may have a role in breast cancer progression (Cancer Res 2001 Dec 1:61 (23): 8448-51). Thus, therapeutic modulation of the expression or function of this gene may be useful for the treatment of these cancers.

  Among tissues with metabolic functions, this gene is expressed at low but significant levels in the pituitary gland, adrenal gland, pancreas, thyroid, skeletal muscle and fetal heart and liver. Expression in these tissues means that this gene product has a role in normal neuroendocrine and metabolic functions, and uncontrolled expression of this gene contributes to neuroendocrine disorders or metabolic diseases such as obesity and diabetes. It may indicate that there is a possibility.

  In addition, this gene is expressed at low levels in all regions of the CNS tested. Steroid treatment is used in many clinical conditions such as Alzheimer's disease (estrogens), menopause-related symptoms (estrogens), multiple sclerosis (glucocorticoids), and spinal cord injury (methylprednisolone). Treatment with an antagonist of the gene product, or a reduction in the level of the gene product, can slow down steroid degradation, reduce the therapeutically effective requirement and, as a result, reduce peripheral side effects. (Holinka CF. Ann NY Acad Sci 2001 Sep; 943: 89-108; Matsumoto T. Spine 2001 Feb 15; 26 (4): 426-30; Gaillard PJ. Neuroreport 2001 Jul 20; 12 (10): 2189-93 ).

  Panel 4.1D Summary: The highest expression of the Ag5926 CG105656-01 gene is found in chronically activated Th1 cells (CT = 34.1). Therefore, therapeutic agents designed with the transcripts are important for the treatment of diseases such as asthma, IBD, psoriasis and arthritis where antigen presentation is reduced or inhibited and T cells are chronically stimulated. Let's go.

  Panel 5 Islet Summary: Ag5926 The expression of this new steroid dehydrogenase-like gene is found exclusively in the liver HepG2 cell line (CT = 33.1). Expression in hepatocytes suggests that the role of this new steroid dehydrogenase may be the same as that of other steroid dehydrogenases in steroid and bile acid metabolism. Therefore, small molecule therapeutics that counter this gene product may be effective in diseases where the expression of this gene is not controlled.

AV. CG105201-1: Hexokinase III splice variant The expression of the gene CG105201-01 was evaluated using the primer-probe set Ag4282 described in Table AVA. The results of RTQ-PCR implementation are shown in Tables AVB, AVC, AVD, AVE and AVF.

Continuation of Table AVB

Continuation of Table AVD

Continuation of Table AVD

Continuation of Table AVE

  AI_Comprehensive Panel_v1.0 Summary: The highest expression of the Ag4282 CG105201-01 gene is detected in samples from rheumatoid arthritis (RA) bone (CT = 27). High to moderate expression of this gene is associated with osteoarthritis (OA) bone and adjacent bone, OA cartilage, OA synovial and OA synovial fluid samples, and RA patient cartilage, bone, It is also observed in samples from membrane and synovial fluid samples. Also, low-level expression is normal lung sample, CODP lung, emphysema, atopic asthma, asthma, allergy, Crohn's disease (normal matching control and disease target), ulcerative colitis (normal matching control and disease) Subject) and samples from psoriasis (normal matched controls and disease subjects). Thus, therapeutic modulation of the gene product may improve symptoms / symptoms associated with autoimmune and inflammatory disorders such as psoriasis, allergies, asthma, inflammatory bowel disease, rheumatoid arthritis and osteoarthritis.

  CNS_Neurodegeneration_v1.0 Summary: Ag4282 This panel confirms that the CG105201-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: Three experiments with the same set of Ag4282 probes and primers agree very well and the highest expression of CG105201-01 gene is found in spleen, colon cancer and bladder (CT = 29-30.8). Therefore, the expression of this gene can be used in this panel to distinguish this sample from other samples and also as a marker for colorectal cancer detection. Furthermore, therapeutic modulation of this gene may be useful for the treatment of colon cancer, allergies, autoimmune diseases, and inflammatory diseases.

Among tissues with metabolic or endocrine functions, this gene is expressed at low to medium levels in the pancreas, adipocytes, adrenal gland, thyroid, skeletal muscle, heart, and liver. Glucose phosphorylation catalyzed by hexokinase is the first triggering reaction in skeletal muscle and adipocyte glucose uptake. Glucose uptake into these tissues is impaired in insulin resistant type 2 diabetes associated with obesity. Pharmacological enhancement of hexokinase activity can be a treatment for the prevention and / or treatment of type 2 diabetes (Jimenez-Chillaron JC, Metabolism, 2002 Jan; 51 (1): 121-6). Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.
Ag4282 One experiment result (experiment 219274018) with this gene is not included. According to the amp plot, there were experimental difficulties in performing this experiment.

  Panel 4.1D Summary: The highest expression of the Ag4282 CG105201-01 gene is detected in resting macrophages (CT = 26). High to moderate expression of this gene is also found in neutrophils, macrophages, monocytes, dendritic cells, two-way MLR, PBMC, and LAK cells. Therefore, the therapeutic regulation of this gene through the use of small molecule drugs is such as Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive pulmonary disease, asthma, emphysema, rheumatoid arthritis, lupus, or psoriasis It may be useful for the treatment of autoimmune and inflammatory diseases.

  General Tumor Screening Panel_v2.4 Summary: Ag4282 The highest expression of this gene is detected in kidney cancer (CT = 31.6). The expression of this gene is found higher in colon, lung, kidney, prostate and metastatic melanoma cancer samples compared to its adjacent normal tissue. Thus, expression of this gene can be used as a marker to detect these cancers, and therapeutic modulation of this gene can be beneficial in the treatment of colon, lung, kidney, prostate and metastatic melanoma cancers .

AW. CG105294-01: Human orthologue of mouse kinesin KIF21B The expression of the gene CG105294-01 was evaluated using the primer-probe set Ag 4283 described in Table AWA. The results of RTQ-PCR execution are shown in Tables AWB, AWC and AWD.

Continuation of Table AWC

Continuation of Table AWD

  CNS_Neurodegenesis_v1.0 Summary: Ag 4283 This panel shows no differential expression of this gene in Alzheimer's disease. This gene encodes a homologue of kinesin, a microtubule-based motor protein involved in organelle transport. APP axonal transport in nerve cells is mediated by binding to kinesin. (Gunewardena S, Neuron 2001 Nov 8:32 (3): 389-401). Kamal et al. Suggest that impaired APP transport increases axon production and precipitates Abeta, thereby disrupting neurotrophic signaling and causing neurodegeneration (Nature 2001). Dec 6; 414 (6864): 643-8). Thus, therapeutic modulation of the expression or function of this gene may be useful for the treatment of Alzheimer's disease or other neurodegenerative disorders.

  General_Screening_Panel_v1.4 Overview: Ag4283 This gene is overexpressed in the fetal brain on this panel compared to expression in other samples (CT = 25.6). Thus, the expression of this gene could be used to distinguish between fetal and adult brain tissue. Rapid organelle transport is required to grow and establish specific structures during neuronal development. Morfini G et al. Showed that kinesin is abundant in growing neurons (Dev Neurosci 2001; 23 (4-5): 364-76). Thus, high levels of expression in the fetal brain suggest that this gene product (kinesin homolog) may be involved in brain development. In addition, this gene is expressed at moderate levels in all regions of the CNS studied. Thus, therapeutic modulation of the expression or function of this gene may be useful for the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Among tissues with metabolic functions, this gene is expressed at low levels in adipocytes, adrenal glands, pancreas, and fetal liver. Expression in these tissues means that this gene product has a role in normal neuroendocrine and metabolic functions, and uncontrolled expression of this gene contributes to neuroendocrine disorders or metabolic diseases such as obesity and diabetes This suggests that it may be.

  Panel 4.1D Summary: Ag 4283 This gene appears to be expressed primarily in hematopoietic cells, with the highest expression being in IL-2 treated LAK cells. This transcript is also found on T cells, particularly chronically activated Th1, Th2 and Tr1 cells. Macrophages, B cells, eosinophils, neutrophils and dendritic cells also express this transcript. The only non-hematopoietic cell type that expresses transcripts at moderate levels is dermal fibroblasts. The thymus and kidney also express the transcript at low levels. Thus, this transcript or the protein it encodes could be used to detect hematopoietic derived cells. In addition, therapeutics designed with the protein encoded by this transcript are important in regulating the function of antigen presenting cells (macrophages and dendritic cells) or T cells, and also asthma, emphysema, psoriasis, arthritis, And important for treatment such as IBD.

AX. CG105334-01: Peptidyl-prolyl cis-trans isomerase The expression of the gene CG105334-01 was evaluated using the primer-probe set Ag4286 described in Table AXA. The results of RTQ-PCR performed are shown in Tables AXB, AXC and AXD.

Continuation of Table AXC

Continuation of Table AXD

  CNS_Neurodegeneration_v1.0 Summary: Ag4286 This panel confirms that the CG105334-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of potential uses of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4286 CG105334-01 gene is detected in the breast cancer T47D cell line (CT = 32.6). Significant expression of this gene is also found in ovarian cancer cell lines. Thus, the expression of this gene can be useful as a marker for detecting the presence of this cancer, and therapeutic modulation of the expression or function of this gene can be useful in the treatment of breast and ovarian cancer.

  In addition, this gene is expressed at low levels in the substantia nigra, thalamus, and cerebral cortex. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4286 CG105334-01 gene is detected in the kidney (Ct = 32.2). In addition, low level expression of this gene is also detected in the thymus. Therefore, the expression of this gene can be used in this panel to distinguish between these two samples and other samples. In addition, therapeutic modulation of the gene product may be beneficial in the treatment of autoimmune and inflammatory diseases that affect the kidney, such as lupus and glomerulonephritis.

AY. CG105503-01: Drosophila melanogaster isolation anxiety protein The expression of the gene CG105503-01 was evaluated using the primer-probe set Ag4289 described in Table AYA. Table AYB shows the results of RTQ-PCR.

Continuation of Table AYB

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4289 CG105503-01 gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Ag4289 CG105503-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: Medium level expression of the Ag4289 CG105503-01 gene is detected only in the kidney (CT = 31.9). Thus, the expression of this gene can be used in this panel to distinguish the kidney from other samples. In addition, small molecule therapeutics designed with the protein encoded by this gene can modulate renal function and are important for the treatment of inflammatory or autoimmune diseases that affect the kidney, such as lupus and glomerulonephritis. possible.

AZ. CG105550-01: Kinesin 12
The expression of the gene CG105550-01 was evaluated using the primer-probe set Ag4291 described in Table AZA. The results of RTQ-PCR performed are shown in Tables AZB and AZC.

Continuation of Table AZB

Continuation of Table AZC

CNS_Neurodegeneration_v1.0 Summary: Ag4291 Expression of this gene is low / undetectable in all of the samples on this panel (CT> 35).
General_Screening_Panel_v1.4 Summary: Ag4291 The highest expression of this gene is found in breast cancer cell lines (CT = 30.9). Medium levels of expression are also found in brain, kidney, ovary and colon cancer cell lines. In addition, higher expression of this gene (CT = 31.4) is detected in fetal kidney compared to expression in adult kidney (CT = 37). Thus, the expression of this gene can be used to distinguish between fetal and adult kidneys and as a marker for these cancers. Expression at significant levels in cancer cell lines and fetal tissues also suggests that this gene product is required for cell growth. Kinesin family members are involved in the assembly and dynamics of mitotic spindles (Blangy A. Cell 1995 Dec 29; 83 (7): 1159-69). The component of the mitotic apparatus is the target of drugs used in cancer therapy. Therefore, based on the homology of this gene with kinesin and its expression in cancer cell lines, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of brain, kidney, breast and colon cancer.

  Panel 4.1D Summary: Ag4291 This gene is expressed at a detectable level in the kidney (CT = 34.4). Therefore, antibodies or small molecule therapies designed with the protein encoded by this gene are important for the treatment of autoimmune and inflammatory diseases that modulate kidney function and affect the kidney such as lupus and glomerulonephritis. obtain.

BA. CG105597-01: Putative 4 repeat ion channel The expression of the gene CG105597-01 was evaluated using the primer-probe sets Ag1090, Ag4292 and Ag536 described in Tables BAA, BAB and BAC. The results of RTQ-PCR performed are shown in Tables BAD, BAE, BAF, BAG, BAH and BAI.

Continuation of Table BAE

Continuation of Table BAF

Continuation of Table BAG

Continuation of Table BAI

  CNS_Neurodegenesis_v1.0 Summary: Ag1090 / Ag4292 Two experiments with different probe and primer sets agree very well. This panel confirms that the CG105597-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4292 CG105597-01 gene, ie the putative ion channel protein, is detected in the whole brain (CT = 27). High expression of this gene is found in all areas of the central nervous system tested, including tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. All mutations known to cause seizure disorders have been found in ion channels, and so far ion channels are the main target of antiepileptic drugs. Thus, this gene can be used to select small molecules that potentially act as small molecule targets for treatment of seizure disorders (including epilepsy) and as anti-epileptic drugs.

  Medium to low levels of expression of this gene are also found in many cancer cell lines derived from pancreatic, lung, breast, ovarian, prostate, squamous cell carcinoma, melanoma and brain cancer. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, lung, breast, ovarian, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at moderate to low levels in adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Panel 1.1 Summary: The highest expression of the Ag536 CG105597-01 gene is detected in the whole brain (CT = 24.5). High expression of this gene is found in all areas of the central nervous system tested, including tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Medium level expression of this gene is also found in many cancer cell lines derived from pancreas, kidney, stomach, lung, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer. Therefore, the expression of this gene can be used as a marker for detecting the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, lung, breast, ovarian, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues having metabolic or endocrine functions, this gene is expressed at moderate to low levels in adipocytes, pancreas, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Panel 4.1D Summary: The highest expression of the Ag4292 CG105597-01 gene is detected in IL-9 treated NCI-H292 (CT = 29.9). In addition, moderate to low level expression of this gene is also observed in endothelial cells, microtubule lung and skin endothelial cells, TNFα + IL-1β treated astrocytes, keratinocytes and NCI-H292 cells presented by HUVEC . Thus, the therapeutic modulation of this gene product is the treatment of multiple sclerosis or other CNS inflammatory diseases, lupus, asthma, emphysema, Crohn's disease, ulcerative colitis, rheumatoid arthritis, osteoarthritis and psoriasis Can be beneficial to.

  Panel 5 Islet Summary: The highest expression of the Ag4292 CG105597-01 gene is detected in adipocytes (CT = 32.4). The novel quadruple ion channel is up-regulated during adipocyte differentiation. The expression profile of the quadruplicate ion channel is very similar to that of PPARg.

  PPARg is necessary and sufficient for adipogenesis (Lee at al., 2002, J Biol Chem 2002 Mar 4, PMID: 11877444 [epub ahead of print]). A novel quadruple ion channel can act downstream of PPARg in promoting adipogenesis. Inhibition of this novel quadruple ion channel delays adipocyte differentiation leading to weight loss. Thus, novel antagonists to the 4-fold ion channel may be beneficial in the treatment of obesity.

  Panel CNS_1 Summary: Ag1090 This panel confirms that the CG105597-01 gene is expressed at low levels in the brain of an independent population. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

BB. CG105627-01 and CG105627-04: C-C chemokine receptor type 5 Expression of genes CG105627-01 and CG105627-04 was evaluated using the primer-probe sets Ag4309 and Ag6626 described in Tables BBA and BBB. The results of RTQ-PCR performed are shown in Tables BBC, BBD, BBE and BBF.

Continuation of Table BBD

Continuation of Table BBE

Continuation of Table BBF

  CNS_Neurodegeneration_v1.0 Summary: Ag4309 This panel confirms that the CG105627-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4309 CG105627-01 gene is detected in the bladder (CT = 30.5). Significant expression of this gene is also observed in tissues having metabolic / endocrine functions such as pancreas, adipocyte, adrenal gland, thyroid, pituitary, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Medium level expression of this gene is also observed in colorectal cancer samples. Therefore, the expression of this gene can be used as a marker to detect the presence of colorectal cancer, and therapeutic regulation of this gene may be beneficial for the treatment of colorectal cancer.

  In addition, the gene is expressed at moderate levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  General_Screening_Panel_v1.6 Summary: The highest expression of the Ag6626 CG105627-01 gene is detected in the bladder (CT = 29.4). Significant expression of this gene is also observed in tissues with metabolic / endocrine functions such as pancreas, adipocytes, adrenal gland, thyroid, pituitary, heart and liver. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Medium level expression of this gene is also observed in colorectal cancer samples. Therefore, the expression of this gene can be used as a marker to detect the presence of colorectal cancer, and therapeutic regulation of this gene may be beneficial for the treatment of colorectal cancer.

  In addition, the gene is expressed at moderate levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: Ag4309 / Ag6626 Two experiments with different probe and primer sets are in very good agreement and CG105627-01 is best expressed in LPS-treated monocytes (CT = 25-25.7) . The CG105627-01 gene encodes a CCR5-like molecule that is a co-receptor for HIV and a chemokine receptor important for moving T cells and mononuclear cells to the site of inflammation. Therefore, a strategy towards targeted CCR5 is important for controlling HIV entry and for the migration of inflammatory cells to the site of tissue injury. Thus, small molecule drugs or antibodies that antagonize the function of this gene have been identified in several types of autoimmune and inflammatory diseases such as lupus, Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive lung Patient symptoms such as disease, asthma, emphysema, rheumatoid arthritis, or psoriasis may be alleviated or eliminated.

BC. CG106074-01: Novel mitochondrial enzyme containing oxidoreductase domain The expression of the gene CG106074-01 was evaluated using the primer-probe set Ag4316 described in Table BCA. The results of RTQ-PCR runs are shown in Tables BCB, BCC and BCD.

Continuation of Table BCC

Continuation of Table BCD

  CNS_Neurodegeneration_v1.0 Summary: Ag4316 This panel confirms that the CG106074-1 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of Ag4316 CG106074-01 gene is detected in kidney cancer TK-10 cell line (CT = 28). Medium level expression of this gene is found in a group of cancer cell lines derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. Furthermore, therapeutic regulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at a medium level in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Interestingly, this gene is expressed at a much higher level in the fetus (CT = 30) compared to adult liver (CT = 35). This observation suggests that the expression of this gene can be used to distinguish between fetal and adult liver. In addition, the relatively overexpression of this gene in fetal liver suggests that the protein product enhances liver growth or development in the fetus and can also act as a regenerative capacity in adults. Therefore, therapeutic modulation of the protein encoded by this gene would be useful in the treatment of liver related diseases.

  In addition, the gene is expressed at moderate levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4316 CG106074-01 gene is detected in basophils (CT = 30.5). The gene is expressed at high to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Is a normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

BD. CG106166-01 and CG106166-02: Novel protein kinases The expression of the genes CG106166-01 and CG106166-02 was evaluated using the primer-probe set Ag4415 described in Table BDA. The results of RTQ-PCR implementation are shown in Tables BDB, BDC, BDD and BDE. Note also that CG106166-02 represents a full-length physical clone of the CG106166-01 gene, validating the prediction of the gene sequence.

Continuation of Table BDB

Continuation of Table BDD

Continuation of Table BDE

  AI_Comprehensive Panel_v1.0 Summary: The highest expression of the Ag4415 CG106166-01 gene is detected in samples from matched controls in patients with psoriasis (CT = 29). Medium level expression of this gene indicates normal and true arthritis / rheumatoid arthritic bone and its adjacent bone, cartilage, synovium and synovial fluid samples, as well as normal lung samples, CODP lungs, emphysema, atopic asthma , Asthma, allergies, Crohn's disease (normal matched controls and disease subjects), ulcerative colitis (normal matched controls and disease subjects), and psoriasis (normal matched controls and disease subjects) Is recognized. Thus, therapeutic modulation of the gene product may improve symptoms / symptoms associated with autoimmune and inflammatory disorders such as psoriasis, allergies, asthma, inflammatory bowel disease, rheumatoid arthritis and osteoarthritis.

  CNS_Neurodegeneration_v1.0 Summary: Ag4415 This panel confirms that the CG106166-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_1.4 Overview: The highest expression of the Ag4415 CG106166-01 gene is detected in the melanoma M14 cell line (CT = 25). Medium to high levels of expression of this gene are also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of pancreatic, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at intermediate levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Interestingly, this gene is expressed at a much higher level in the fetus (CT = 29.7) compared to adult liver (CT = 34.8). This observation suggests that the expression of this gene can be used to distinguish between fetal and adult liver. In addition, the relatively overexpression of the gene in fetal liver suggests that the gene product enhances liver growth or development in the fetus and can also act as a regenerative capacity in adults. Therefore, therapeutic modulation of the protein kinase encoded by this gene would be useful in the treatment of liver related diseases.

  In addition, the gene is expressed at moderate levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Thus, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4415 CG106166-01 gene is detected in activated secondary Th2 cells (CT = 29). The gene is expressed at high to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Normal tissue presented by This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

BE. CG106773-01: Calmodulin-dependent protein kinase II-δ
The expression of the gene CG106773-01 was evaluated using the primer-probe set Ag4336 described in Table BEA. The results of RTQ-PCR runs are shown in Tables BEB, BEC, BED and BEE.

Continuation of Table BEC

Continuation of Table BED

  CNS_Neurodegeneration_v1.0 Summary: Ag4336 This panel confirms that the CG106773-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4336 CG106773-01 gene is detected in the breast cancer BT549 cell line (CT = 25). Medium to high levels of expression of this gene are also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. The CG106773-01 gene encodes a splice variant of the calcium / calmodulin-dependent protein kinase type II delta chain (CaMKII). CaMKII has been implicated in various neuronal and non-neuronal cell functions such as control of cell proliferation. A splice variant of CaMKII has been shown to be differentially expressed on tumor cells, particularly neuroblastoma and breast cancer cells (Tombes RM, Krystal GW., 1997, Biochim Biophys Acta 1355 (3): 281- 92, PMID: 9060999). Thus, therapeutic modulation of this gene through the use of small molecule drugs may be beneficial for the treatment of neuroblastoma and breast tumors.

  Among tissues with metabolic or endocrine functions, this gene is expressed at a medium level in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. CaMK-II is highly expressed in pancreatic islets and is known to be associated with insulin-secreting vesicles. Moreover, suppression of CaMKII disrupts insulin secretion and insulin gene expression (Rochlitz et al., 2000, Diabetologia 43 (4): 465-73, PMID: 10819240). Activation of CaMKII and associated phosphorylation of synapsin I contribute to insulin secretion from pancreatic β cells (Tabuchi et al., 2000, Endocrinology 141 (7): 2350-60, PMID: 10875234). Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Calmodulin (CaM) is a major Ca 2+ binding protein in the brain and plays an important role in responding to neurons against changes in intracellular Ca 2+ concentration. Calmodulin modulates many Ca2 + -dependent enzymes and is involved in related cellular functions. Among the different CaM-binding proteins, Ca2 + / CaM dependent protein kinase II and phosphatase calcineurin are particularly important in the brain because of their abundance and involvement in many neuronal functions (Sola et al. , 2001, Int J Biochem Cell Biol 33 (5); 439-55, PMID: 11331200). In addition, α-CaMKII knockout mice and transgenic mice expressing mutant forms of CaMKII clearly show that CaMKII plays a prominent role in hippocampal LTP and hippocampal-dependent memory (Fukunaga K, Miyamoto E , 2000, Neurosci Res 38 (1): 3-17, PMID: 10997573). Therefore, therapeutic modulation of this gene product may be useful in the treatment of neuronal disorders such as memory loss, Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of the Ag4336 CG106773-01 gene is detected in IL-9 treated lung fibroblasts (CT = 27.3). This gene is expressed at high to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Normal tissue presented by This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, And can improve symptoms in patients with autoimmune and inflammatory diseases such as osteoarthritis.

  Panel 5 Islet Summary: The highest expression of the Ag4336 CG106773-01 gene is detected in skeletal muscle (CT = 29.5). In addition, moderate expression of this gene is also found in islet cells, adipocytes, skeletal muscle, uterus, placenta, kidney and small intestine. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes. See panel 1.4 for further discussion and discussion of the use of this gene.

BF. CG108211-01: ECT2
The expression of the gene CG108211-01 was evaluated using the primer-probe set Ag4353 described in Table BFA.

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4353 CG108211-01 gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: Expression of the Ag4353 CG108211-01 gene is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Summary: Expression of the Ag4353 CG108211-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

BG. CG108351-01: Thioredoxin reductase TR2
The expression of gene CG108351-01 was evaluated using the primer-probe set Ag4354 described in Table BGA. The results of RTQ-PCR are shown in Tables BGB and BGC.

Continuation of Table BGB

Continuation of Table BGC

  CNS_Neurodegenesis_v1.0 Summary: Expression of Ag4354 CG108351-01 gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4354 CG108351-01 gene is detected in the CNS cancer (star) SNB-75 cell line (CT = 27.9). Medium to high level expression of this gene is also found in a group of cancer cell lines derived from pancreas, stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. The CG108351-01 gene encodes thioredoxin reductase (TR). TR is selenocysteine containing a flavin enzyme that catalyzes the NADPH-dependent reduction of the redox protein thioredoxin. Thioredoxin is overexpressed in many human tumors. Experimental studies have shown that thioredoxin is involved in the growth and transformation phenotype of human cancer cells in part. Thus, thioredoxin reductase provides an attractive target for the development of anticancer drugs to control the activity of the thioredoxin system (Engman et al., 1997, Anticancer Res 17 (6D): 4599-605, PMID: 9494575). Therefore, therapeutic regulation of TR expression or function encoded by this gene is effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer obtain.

  Among tissues with metabolic or endocrine functions, this gene is expressed at a medium level in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, such as tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Panel 4.1D Summary: The highest expression of Ag4354 CG108351-01 gene is detected in kidney and IFNγ treated NCI-H292 (CT = 31.7). Low to moderate expression of this gene is due to endothelial cells derived from the skin and lung epithelium, epithelial cells, astrocytes, lung and skin fibroblasts, keratinocytes, basophils, dendritic cells, activation It is found in lymphocytes, activated primary and secondary T cells, and normal tissues presented by the lungs and thymus. Therefore, therapeutic regulation of this gene may be useful for the treatment of asthma, allergies, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, and osteoarthritis.

BH. CG108462-01: Novel FKBP-type peptidyl-prolyl cis-trans isomerase The expression of the gene CG108462-01 was evaluated using the primer-probe set Ag4364 described in Table BHA. The results of RTQ-PCR performed are shown in Tables BHB, BHC and BHD.

Continuation of Table BHC

Continuation of Table BHD

  CNS_Neurodegeneration_v1.0 Summary: Ag4364 This panel confirms that this gene is expressed at low levels in the brain of an independent population. This gene is found slightly up-regulated in the temporal cortex of Alzheimer patients. Therefore, neuronal cell death can be reduced by therapeutic regulation of the expression or function of this gene, which is useful for the treatment of this disease.

  General_Screening_Panel_v1.4 Summary: Ag4364 The highest expression of this gene is detected in brain cancer cell lines (CT = 27). A significant level of expression of this gene is also observed in a group of brain cancer cell lines. The gene is widely expressed in this panel, with moderate expression also found in brain, colon, stomach, lung, breast, ovary, pancreas and melanoma cancer cell lines. This expression profile suggests a role for this gene product in cell survival and proliferation. The therapeutic modulation of this gene product may be useful for the treatment of cancer.

  Among tissues with metabolic functions, this gene is expressed at low to significant levels in the pituitary gland, adipocytes, adrenal glands, pancreas, thyroid, and adult and fetal skeletal muscles, heart and liver. Widespread expression in these tissues indicates that this gene product has a role in normal neuroendocrine and metabolic functions, and that unregulated expression of this gene is associated with neuroendocrine disorders or metabolic diseases such as obesity and diabetes. It suggests that it can contribute.

  This gene is expressed at intermediate levels in CNS such as hippocampus, thalamus, substantia nigra, tonsils, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Panel 4.1D Summary: Ag4365 The highest expression of this gene is found in resting monocytes (CT = 27.9). The gene is expressed at high to moderate levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Is a normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.4, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change functions associated with these cell types, such as asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

BI. CG108723-01, CG108723-02 and CG108723-03: Sodium and chloride-dependent GABA transporter The expression of the genes CG108723-01, CG108723-02 and CG108723-03 is expressed in the primer-probe set Ag4408 described in Tables BIA, BIB and BIC , Ag5971 and Ag6397. Note that probe Ag6397 is specific for mutant CG108723-03. It should also be noted that CG108723-02 represents a full-length physical clone of the CG108723-01 gene, validating the prediction of the gene sequence. The results of RTQ-PCR are shown in Table BID.

CNS_Neurodegenesis_v1.0 Summary: Ag4408 Two experiments with the same probe and primer set agree well. This panel confirms that the CG108723-01 gene is expressed at low levels in the brain of an independent population. This gene is found slightly down-regulated in the temporal cortex of Alzheimer's disease patients. Thus, upregulation of this gene or its protein, or treatment with a specific agonist for this receptor, is useful in reversing dementia, memory loss, and neuronal cell death associated with the disease.
Ag6397 This probe is specific for CG108723-03 and the expression of this gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.6 Summary: Ag6397 CG108723-03 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 4.1D Overview: Ag6397 CG108723-03 gene expression is low / undetectable across all samples on this panel (CT> 35).

Panel 5 Islet Summary: Ag4408 Ag4408 CG108723-01 gene expression is low / undetectable across all samples on this panel (CT> 35).
Ag6397 This probe is specific for CG108723-03 and the expression of this gene is also low / undetectable across all the samples on this panel (CT> 35).

  Panel 5D Summary: Ag5971 CG108723-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel CNS_1 Summary: The results of one experiment with the Ag4408 CG108723-01 gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

BJ. CG108870-01: HS1 binding protein 3
The expression of gene CG108870-01 was evaluated using the primer-probe set Ag4382 described in Table BJA. The results of RTQ-PCR are shown in Tables BJB, BJC and BJD.

Continuation of Table BJC

Continuation of Table BJD

  CNS_Neurodegeneration_v1.0 Summary: Ag 4382 This panel confirms that the CG108870-01 gene is expressed at low levels in the brain of an independent population. This gene appears to be slightly up-regulated in the temporal cortex of Alzheimer's disease patients. Therefore, therapeutic modulation of the expression or function of this gene reduces neuronal cell death and may be useful in the treatment of this disease.

  General_Screening_Panel_1.4 Summary: Ag4382 The highest expression is detected in breast cancer cell lines (CT = 27). High to medium levels of expression are found in cell lines derived from brain, colon, liver, lung, breast, ovary, and skin cancers. In addition, this gene is expressed at much higher levels in the fetal lung (CT = 30) than in the adult lung (CT = 33). Thus, the expression of this gene can be used to distinguish whether this tissue is of fetal or adult origin. High levels of expression in fetal tissues and cancer cell lines suggests that this gene has a role in cell survival and proliferation. This therapeutic modulation of the gene product may be useful in the treatment of cancer.

  Among tissues with metabolic functions, this gene is expressed at moderate to low levels in the pituitary gland, adipocytes, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. Extensive expression in these tissues indicates that this gene product has a role in normal neuroendocrine and metabolic functions, and that unregulated expression of this gene is associated with neuroendocrine disorders or metabolic diseases such as obesity and diabetes. It suggests that it can contribute.

  In addition, this gene is expressed at a medium level in CNS such as hippocampus, thalamus, substantia nigra, tonsils, cerebellum and cerebral cortex. Therefore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

  Panel 4.1D Summary: Ag4382 The highest expression of this gene is found in activated skin fibroblasts (CT = 29.7). The gene is expressed at moderate to low levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells are members of the T cell, B cell, endothelial cell, and peripheral blood mononuclear cell families, as well as epithelial and fibroblast types from the lung and skin, and normal tissues presented by the colon, lung, thymus and kidney Etc. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.5, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change functions associated with these cell types, such as asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

BK. CG109487-01: voltage-dependent anion channel 2
The expression of gene CG109487-01 was evaluated using the primer-probe set Ag4385 described in Table BKA. The results of RTQ-PCR performed are shown in Tables BKB and BKC.

Continuation of Table BKB

Continuation of Table BKC

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4385 CG109487-01 gene is low / undetectable across all samples on this panel (CT> 35).

General_Screening_Panel_v1.4 Summary: The highest expression of Ag4385 CG109487-01 gene is detected in the trachea (CT = 32.9). Therefore, therapeutic regulation of this gene may be useful in the treatment of trachea-related diseases.
In addition, significant expression of this gene is also found in many cancer cell lines, including CNS, colon, stomach, and breast cancer cell lines. Therefore, the expression of this gene can be used as a diagnostic marker to detect the presence of these cancers, and the therapeutic modulation of this gene through the use of small molecule targets has been demonstrated in the CNS, colon, stomach, and breast. May be beneficial in the treatment of cancer.

  Panel 4.1D Summary: The highest expression of the Ag4385 CG109487-01 gene is detected in the kidney (CT = 30.4). Therefore, the expression of this gene can be used to distinguish the kidney from other samples used in this panel. In addition, therapeutic regulation of this gene may be beneficial in the treatment of autoimmune or inflammatory diseases that affect the kidney, such as lupus and glomerulonephritis.

  Medium to low level expression of this gene is also observed in thymus, basophils, eosinophils and cytokine-treated LAK cells. Therefore, therapeutic modulation of this gene product may be beneficial in the treatment of asthma, allergies, irritable reactions, inflammatory bowel disease, viral infections and autoimmune diseases.

  Panel CNS_1 Summary: Expression of the Ag4385 CG109487-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  Panel CNS_1.1 Summary: Expression of the Ag4385 CG109487-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

BL. CG109594-01: Phospholipase The expression of the gene CG109594-01 was evaluated using the primer-probe set Ag4393 described in Table BLA. The results of RTQ-PCR are shown in Table BLB.

Continuation of table BLB

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4393 CG109594-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4393 CG109594-01 gene is detected only in the lung cancer A549 cell line (CT = 26.3). Therefore, the expression of this gene could be used on this panel to distinguish this sample from other samples and as a marker to detect the presence of lung cancer. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of lung cancer.

  Panel 4.1D Summary: Expression of the Ag4393 CG109594-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

  Panel 5 Islet Summary: The results of one experiment with the Ag4393 CG109594-01 gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

  General Tumor Screening Panel_v2.4 Summary: Expression of the Ag4393 CG109594-01 gene is low / undetectable across all samples on this panel (CT> 35).

BM. CG109733-01: Guanyl kinase-like gene Expression of CG109733-01 was evaluated using the primer-probe set Ag4401 described in Table BMA.

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag4401 CG109733-01 gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.4 Summary: The results of one experiment with the Ag4401 CG109733-01 gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

  Panel 4.1D Overview: Ag4401 CG109733-01 gene expression is low / undetectable across all of the samples on this panel (CT> 35).

  Panel CNS_1 Summary: Ag4401 CG109733-01 gene expression is low / undetectable across all of the samples on this panel (CT> 35).

  Panel CNS_1.1 Summary: Expression of the Ag4401 CG109733-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

BN. CG109835-01: GTP-binding protein REM 2
The expression of gene CG109835-01 was evaluated using the primer-probe set Ag4385 described in Table BNA. The results of RTQ-PCR are shown in Tables BNB, BNC, BND, BNE and BNF.

Continuation of Table BNC

Continuation of Table BND

Continuation of Table BNE

Continuation of Table BNF

  CNS_Neurodegeneration_v1.0 Summary: Ag4405 This panel confirms that the CG109835-01 gene is expressed at low levels in the brain of an independent population. However, differential expression of this gene was not detected between the postmortem brain of Alzheimer's disease and the non-demented control of this experiment. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  General_Screening_Panel_v1.4 Summary: The highest expression of the Ag4405 CG109835-01 gene is detected in the cerebellum (28.5). High to moderate expression of this gene is found in all areas of the central nervous system tested, including tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. The CG109835-01 gene encodes a homologue of the rat GTP binding protein REM 2 (REM2). Rat rem 2 was identified based on homology with the Rem, Rad, Gem and Kir (RGK) families of Ras-related small GTP binding proteins. Rem2 mRNA was detected in rat brain and kidney (Finlin et al., 2000, Biochem J 347 Pt 1: 223-31, PMID: 10727423). Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  Also, moderate to low level expression of this gene is found in a group of cancer cells derived from stomach, colon, lung, kidney, breast, ovary, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. Furthermore, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues having metabolic or endocrine functions, this gene is expressed at a medium to low level in pancreas, adipocyte, pituitary, skeletal muscle, heart, fetal liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  Interestingly, this gene is expressed at a much higher level in the fetus (CT = 32.7) compared to adult liver (CT = 40). This observation suggests that the expression of this gene can be used to distinguish between fetal and adult livers. In addition, the relatively overexpression of the gene in fetal liver suggests that the protein product enhances liver growth or development in the fetus and can also act as a regenerative capacity in adults. Therefore, therapeutic modulation of the rem2 protein encoded by this gene would be useful in the treatment of liver related diseases.

  Panel 4.1D Summary: The highest expression of the Ag4405 CG109835-01 gene is detected in resting neutrophils. Medium level expression of this gene is also observed in TNFα + LPS treated neutrophils. Therefore, expression of this gene can be used in this panel to distinguish neutrophils from other samples. Also, low level expression of this gene is found in thymus, IL-4 treated dermal fibroblasts, PMA / ionomycin treated basophils, dendritic cells, monocytes, CD40L and IL-4 activated B lymphocytes, NK cells. , LAK cells, anti-CD95CH11 treated secondary Th1 / Th2 / Tr1 cells, and CD4 lymphocytes. Therefore, small molecule drugs that antagonize the function of this gene product have several types of autoimmune and inflammatory diseases such as lupus, Crohn's disease, ulcerative colitis, multiple sclerosis, chronic obstructive lung Patient symptoms such as disease, asthma, emphysema, rheumatoid arthritis, or psoriasis may be alleviated or eliminated.

  Panel CNS_1 Summary: Ag4405 This panel confirms that the CG109835-01 gene is expressed at low levels in the brain of an independent population. See Panel 1.4 for a discussion of the potential use of this gene in the treatment of central nervous system disorders.

  Panel CNS_1.1 Summary: Ag4405 This panel confirms that the CG109835-01 gene is expressed at low levels in the brain of an independent population. See panel 1.4 for a discussion of potential uses of this gene in the treatment of central nervous system disorders.

BO. CG110114-01 and CG110114-02: aldose reductase related proteins The expression of the genes CG110114-01 and CG110114-02 was evaluated using the primer-probe set Ag5947 described in Table BOA. The results of RTQ-PCR are shown in Table BOB. Note also that CG110114-02 represents a full-length physical clone of the CG110114-01 gene, validating the prediction of the gene sequence.

Continuation of Table BOB

  CNS_Neurodegeneration_v1.0 Summary: Expression of the Ag5947 CG110114-01 gene is low / undetectable across all samples on this panel (CT> 35).

  General_Screening_Panel_v1.5 Summary: The highest expression of Ag5947 CG110114-01 gene is detected in lung cancer NCI-H460 cell line (CT = 32.6). In addition, medium level expression of this gene is also observed in lung cancer A549 and kidney cancer A498 cell lines. Therefore, the expression of this gene can be used on this panel to distinguish these samples from other samples and as a diagnostic marker for detecting these cancers. Furthermore, therapeutic regulation of this gene through the use of small molecule drugs may be beneficial for the treatment of these cancers.

  The CG110114-01 gene encodes a homologue of aldo-keto reductase. Aldo-keto reductase (AKR) is a family of monomeric oxidoreductases with a molecular weight range of 35-40 kDa, currently over 60 types. The enzyme is expressed in a variety of tissues and catalyzes the NADP-dependent reduction of various aliphatic and aromatic aldehydes and ketones. Members of the AKR family are overexpressed in hepatocellular carcinoma (HCC) and have been shown to contribute to drug resistance (Scuric et al., 1998, Haptology 27 (4); 943-50, PMID: 9537432; Lee et al., 2001, Anticancer Drugs 12 (2): 129-32, PMID: 11261885).

  Two members of the AKR family, aldehyde reductase (AKRIA) and aldose reductase (AKRIB), have been extensively studied as being associated with diabetic complications. AKRIB is up-regulated in the case of hyperlipidemia, with increased activity of the sorbitol pathway and non-enzymatic glycation of proteins, followed by various tissue damage. Unlike other AKR family members that are expressed throughout, recently kidney-specific oxidoreductases have been described as being expressed exclusively in proximal tubules. Although not homologous to other AKR members, it binds NADPH with high affinity and is up-regulated in mice in streptozotocin-induced diabetes. It is also developmentally controlled and appears to selectively regulate renal tubule formation during the fetal period (Wallner et al., 2001, Ren Fail 23 (3-4): 311-20, PMID: 11499547). Therefore, the AKR protein encoded by this gene may have a functional association with diabetic complications, and therapeutic regulation of this gene may be beneficial in the treatment of diabetes.

Panel 4.1D Summary: Expression of the Ag5947 CG110114-01 gene is low / undetectable across all of the samples on this panel (CT> 35).
Panel 5 Isle Summary: The results of one experiment with the Ag5947 CG110114-01 gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

BP. CG110123-01: Kinesin family member C2 (Kinesin motor protein)
The expression of gene CG110123-01 was evaluated using the primer-probe set Ag4409 described in Table BPA. The results of RTQ-PCR execution are shown in Tables BPB, BPC and BPD.

Continuation of Table BPC

Continuation of Table BPD

  CNS_Neurodegeneration_v1.0 Summary: Ag4409 This panel confirms that this gene is expressed at low levels in the brain of an independent population. This gene is found to be down-regulated in the temporal cortex of Alzheimer's disease patients. This gene encodes a homolog of the microtubule-based motor protein kinesin involved in organelle transport, such as APP axonal transport in neurons. (Gunewardena S, Neuron 2001 Nov 8; 32 (3): 389-401). Kamal et al. Suggest that impaired APP transport increases axon production and precipitates Abeta, thereby disrupting neurotrophic signaling and causing neurodegeneration (Nature 2001). Dec 6; 414 (6864): 643-8). Therefore, based on the expression pattern of this kinesin homologue, up-regulation of this gene or its protein, or treatment with a specific agonist for this receptor, is associated with dementia, memory loss, and neuronal cell death associated with the disease. This is useful when reversing.

  General_Screening_Panel_v1.4 Summary: Ag4409 This gene is ubiquitously expressed in this panel and is most expressed in breast cancer cell lines (CT = 28.8). A moderate level of expression is also observed in all cancer cell lines on this panel. Therefore, the expression of this gene could be used as a cancer marker. Expression at significant levels in cancer cell lines suggests that this gene product, a kinesin homolog, is required for cell growth. Kinesin family members are involved in mitotic spindle assembly and dynamics (Blangy A. Cell 1995 Dec 29; 83 (7): 1159-69). The component of the mitotic apparatus is the target of drugs used for cancer treatment. Therefore, based on the homology of this gene with kinesin and its expression in cancer cell lines, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of cancer.

  In addition, a group of significant expression is detected in all samples from the CNS. The kinesin C elegance ortholog has been shown to be involved in axonal transport. In addition, null mutants exhibit several behavioral defects (Jamain S. Genomics 2001 May 15; 74 (1): 36-44). Thus, this therapeutic modulation of kinesin expression or function may be effective in the treatment of neurological disorders.

  Panel 4.1D Summary: Ag4409 The expression of this gene is low but significantly spread at a significant level in this panel. The highest expression is found in the kidney (CT = 31.1). This gene also includes T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, as well as epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and It is also expressed in normal tissues presented by the kidney. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of the general screening panel v1.5, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change the functions associated with these cell types, asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

BQ. CG110132-01: Novel MMKIF17 protein (kinesin motor protein)
The expression of gene CG110132-01 was evaluated using the primer-probe set Ag4410 described in Table BQA. The results of RTQ-PCR execution are shown in Tables BQB, BQC and BQD.

Continuation of Table BQC

Continuation of Table BQD

  CNS_Neurodegeneration_v1.0 Summary: Ag4410 This panel shows no differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_1.4 Summary: Ag4410 The expression of this gene appears to be brain specific and the expression of this gene is highest in the cerebellum (CT = 28.9). Medium levels of expression are found throughout the CNS, eg, in the hippocampus, cerebral cortex, substantia nigra, thalamus, and tonsils. The kinesin C elegance ortholog has been shown to be involved in axonal transport. In addition, null mutants exhibit several behavioral defects (Jamain S. Genomics 2001 May 15; 74 (1): 36-44). Thus, this therapeutic modulation of kinesin expression or function may be effective in the treatment of neurological disorders.

  Significant levels of expression are also found in breast and lung cancer cell lines. Kinesin family members are involved in the assembly and dynamics of mitotic spindles, which are targets for drugs used in cancer treatment (Blangy A. Cell 1995 Dec 29; 83 (7): 1159-69) . Therefore, based on the homology of this gene with kinesin and its expression in cancer cell lines, therapeutic modulation of the expression or function of this gene product may be useful in the treatment of breast and lung cancer.

  Panel 4.1D Summary: Ag4410 The highest expression of this gene is found in the kidney (CT = 30.1). Therefore, the expression of this gene could be used on this panel to distinguish kidney samples from other samples and as a marker for kidney tissue. This prominent expression in the kidney suggests that this kinesin may have a role in renal function, such as vesicular transport based on microtubules of ion channels and transporters to and from the apex of cell membranes. To do. The putative membrane transport activity of this protein can contribute to certain functions of the kidney, such as maintaining ion channels and transporter composition in the apical membrane. In addition, defects in microtubule-based transporters can contribute to kidney disease. (Hamm-Alvarez SF, Physiol Rev 1998 Oct; 78 (4): 1109-29). Therefore, small molecule therapeutics designed with the protein encoded by this gene can modulate renal function and can be used to treat inflammatory or autoimmune diseases that affect the kidney, such as lupus and glomerulonephritis. is important.

BR. CG110160-01 and CG110160-02: Novel globin-like proteins The expression of the genes CG110160-01 and CG110160-02 was evaluated using the primer-probe set Ag4411 described in Table BRA. The results of performing RTQ-PCR are shown in Tables BRB, BRC and BRD. Note also that CG110160-02 represents a full-length physical clone of the CG110160-01 gene, validating the prediction of the gene sequence.

Continuation of Table BRC

Continuation of Table BRD

  CNS_Neurodegeneration_v1.0 Summary: Ag4411 This panel does not show differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: Ag4411 The highest expression of this gene is detected in brain cancer cell lines (CT = 24.4). High levels of expression are also found in cell lines derived from ovarian cancer and melanoma. This protein is homologous to an astrocyte activation-related protein and is a novel peroxidase that can act as an antifibrosis that traps peroxides in the liver (Kawada N, J Biol Chem 2001 Jul 6: 276 (27 ): 25318-23). This gene can therefore be used on this panel to distinguish these samples from other samples. Furthermore, therapeutic modulation of the expression or function of this gene may be useful in the treatment of these cancers.

  Among tissues with metabolic functions, this gene is expressed at moderate to low levels in the pituitary gland, adipocytes, adrenal gland, pancreas, thyroid, and adult and fetal skeletal muscle, heart, and liver. Widespread expression in these tissues indicates that this gene product has a role in normal neuroendocrine and metabolic functions, and that unregulated expression of this gene is associated with neuroendocrine disorders or metabolic diseases such as obesity and diabetes. It suggests that it can contribute.

  In addition, this gene is expressed at a high level in the cerebellum, and at a medium level in all CNS such as hippocampus, thalamus, substantia nigra, tonsils and cerebral cortex. Therefore, therapeutic regulation of the expression or function of this gene can be used to treat neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, seizures and epilepsy, as well as autism and ataxia May be useful in the treatment of pathologies affecting the cerebellum.

  Panel 4.1D Summary: Ag4411 The highest expression of this gene is found in keratinocytes (CT = 30). Medium levels of expression are found in activated lung fibroblasts, dermal fibroblasts and bronchial epithelium, treated and untreated small airway epithelium and coronary SMC, and normal lung. Therefore, the expression of this gene could be used on this panel to distinguish keratinocytes from other samples. Lung and skin-derived cells, or the expression of genes therein, indicate that this gene is normal, as well as pathological and inflammatory lung and skin disorders such as chronic obstructive pulmonary disease, asthma, allergies, qi It may be involved in tumors, psoriasis and wound healing.

BS. CG110350-01: Photoreceptor outer segment all-trans-retinol dehydrogenase The expression of the gene CG110350-01 was evaluated using the primer-probe set Ag5915 described in Table BSA.

  General_Screening_Panel_v1.5 Summary: Ag5915 CG110350-01 gene expression is low / undetectable across all samples on this panel (CT> 35).

  Panel 5 Islet Summary: Expression of the Ag5915 CG110350-01 gene is low / undetectable across all of the samples on this panel (CT> 35).

BT. CG59693: trans-1,2-dihydrobenzene-1,2-diol dehydrogenase The expression of NOV70 mutants CG59693-01 and CG59693-02 was evaluated using the primer-probe set Ag3562 described in Table BTA. The results of RTQ-PCR performed on CG59693-02 are shown in Tables BTB, BTC, BTD, BTE, BTF, BTG, BTH and BTI. The results of RTQ-PCR performed on CG59693-01 are shown in Tables BTJ, BTK, BTL and BTM.

Continuation of Table BTC

Continuation of Table BTD

Continuation of Table BTE

Continuation of Table BTF

Continuation of Table BTG

Continuation of Table BTH

Continuation of Table BTJ

Continuation of Table BTJ

Continuation of Table BTK

Continuation of Table BTK

Continuation of Table BTL

Continuation of Table BTL

  CNS_Neurodegeneration_v1.0 Summary: Ag3562 This panel confirms that the CG59969-02 gene is expressed at low levels in the brain of an independent population. This gene, when analyzed by ANCOVA (p = 0.002), is found up-regulated in the temporal cortex of Alzheimer's disease patients. Therefore, treatment with antagonists or agonists can prevent or delay the progression of Alzheimer's disease.

  General_Screening_Panel_v1.4 Summary: Ag3562 The highest expression of this gene is detected in the lung cancer A549 cell line (CT = 20.01). High expression of this gene is also found in a group of cancer cell lines derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Thus, the expression of this gene can be used as a marker to detect the presence of these cancers. Moreover, therapeutic modulation of the expression or function of this gene may be effective in the treatment of stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma and brain cancer.

  Among tissues with metabolic or endocrine functions, this gene is expressed at medium to high levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

  In addition, the gene is expressed at high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Therefore, therapeutic modulation of this gene product may be useful in the treatment of central nervous system disorders such as Alzheimer's disease, Parkinson's disease, epilepsy, multiple sclerosis, schizophrenia and depression.

  General_screening_panel_v1.6 Summary: The highest expression of the Ag3562 CG59693-02 gene is detected in the lung cancer A549 cell line (CT = 20.7). High expression of this gene is found in a group of cancer cell lines derived from stomach, colon, lung, kidney, breast, ovary, prostate, squamous cell carcinoma, melanoma, and brain cancer. Among tissues with metabolic or endocrine functions, this gene is expressed at medium to high levels in the pancreas, adipocytes, adrenal gland, thyroid, pituitary, skeletal muscle, heart, liver and gastrointestinal tract. In addition, the gene is expressed at high levels in all areas of the central nervous system tested, including the tonsils, hippocampus, substantia nigra, thalamus, cerebellum, cerebral cortex, and spinal cord. Since this pattern is consistent with the expression profile of General_Screening_Panel_v1.4, see Panel 1.4 for a discussion of possible uses of this gene.

HASS Panel v1.0 Summary: Ag3562 CG59693-02 gene expression is not increased in this panel by hypoxia, acidic or serum starvation environment in breast, bladder, pancreas and prostate cell lines.
However, in glioblastoma / astrocytoma cell lines, when these cells are subjected to an acidic environment, their expression increases (maximum expression U87-MGF11; CT = 23.96), which is a brain cancer. This suggests that expression may be up-regulated in the acidic region. Moderate to low expression is shown in this panel in 2 of the 5 gliomas and in 2 of the 4 medulloblastomas. The therapeutic modulation of this gene product by small molecule drugs may be useful for the treatment of brain cancer.

  Tumor_cell_strain_screening_panel v3.1 Summary: The highest expression of the Ag3562 CG59693-02 gene is detected in lung carcinoid samples (CT = 21.7). High to moderate expression of this gene is found in many cancer samples, including tongue, breast, prostate, melanoma, bone marrow, bladder, pancreas, kidney, lymphoma, ovary, cervix, uterus, stomach, lung and brain cancer. Is also accepted. Thus, therapeutic modulation of this gene through the use of small molecule drugs may be beneficial for the treatment of these cancers.

  Panel 2D Summary: The highest expression of the Ag3562 CG59693-02 gene is detected in lung cancer (CT = 23.5). High expression of this gene is found in many lung cancer samples. The expression of this gene is higher in cancer samples compared to the corresponding adjacent control sample. Therefore, expression of this gene can be used as a marker to detect the presence of lung cancer, and therapeutic modulation of this gene through the use of small molecule drugs may be useful for the treatment of lung cancer.

  High to moderate expression of this gene is also found in many cancer samples including cancers of the colon, stomach, ovary, liver, breast, thyroid, kidney, and prostate. Thus, therapeutic modulation of this gene through the use of small molecule drugs may be beneficial for the treatment of these cancers.

  Panel 4.1D Summary: The highest expression of the Ag3562 CG59693-02 gene is detected in IL-4 treated dermal fibroblasts (CT = 25.2). This gene is expressed at moderate to low levels in a wide range of cell types that are meaningful for healthy and diseased immune responses. These cells include T cells, B cells, endothelial cells, macrophages / monocytes, and members of the peripheral blood mononuclear cell family, and epithelial and fibroblast types from the lung and skin, and the colon, lung, thymus and kidney. Is a normal tissue. This ubiquitous expression pattern suggests that the gene product may be involved in the homeostasis process of these and other cell types and tissues. This pattern is consistent with the expression profile of General_Screening_Panel_v1.5, suggesting a role for this gene product in cell survival and proliferation. Therefore, by modulating this gene product with a functional therapeutic agent, it is possible to change functions associated with these cell types, such as asthma, allergy, inflammatory bowel disease, lupus, psoriasis, rheumatoid arthritis, It is possible to improve the symptoms of patients with autoimmune diseases such as osteoarthritis and inflammatory diseases.

  Panel 5 Islet Summary: The highest expression of the Ag3562 CG59693-02 gene is detected in islet cells (Bayer patient 1) (CT = 25.3). High to moderate expression of this gene is also found in adipocytes, skeletal muscle, placenta, uterus, liver, heart, small intestine and kidney. Therefore, this therapeutic modulation of gene activity may prove useful in the treatment of endocrine / metabolism related diseases such as obesity and diabetes.

BU. CG93541-01, CG93541-03, CG93541-04, CG93541-05 and CG93541-06: autotaxin-t (atx-t)
The expression of the genes CG93541-01, CG93541-03, CG93541-04, CG93541-05 and CG93541-06 was evaluated using the primer-probe set Ag3857 described in Table BUA. The results of RTQ-PCR implementation are shown in Tables BUB, BUC, BUD, BUE, BUF and BUG.

Continuation of Table BUC

Continuation of Table BUD

Continuation of Table BUD

  CNS_Neurodegenesis_v1.0 Summary: Ag3857 This panel shows no differential expression of this gene in Alzheimer's disease. However, this expression profile confirms the presence of this gene in the brain. See panel 1.4 for a discussion of the use of this gene in the central nervous system.

  General_Screening_Panel_v1.4 Summary: Ag3857 The results of one experiment with this gene are not included. According to the amp plot, there were experimental difficulties in performing this experiment.

  General_Screening_Panel_v1.5 Summary: The highest expression of Ag3857 CG93541-01 gene is detected in the spinal cord (CT = 25.1). This gene is expressed in the high region of the CNS in general. Therefore, the expression of this gene can be used on this panel to distinguish between brain-derived samples and other samples and as a brain tissue marker. This gene is homologous to autotaxin and is rich in the spinal cord and brain of the rat and may be involved in oligodendrocyte function (Fuss B. J Neurosci 1997 Dec 1; 17 (23): 9095-103 ). Thus, the strong association of this gene with the CNS and the homology with autotaxin makes therapeutic regulation of this gene or gene product useful for the treatment of neurological diseases, particularly demyelinating diseases such as multiple sclerosis. Suggests that it can be.

  In addition, this gene is expressed at a much higher level in the fetus (CT = 26.5-27.5) than in adult lung and liver tissues (CT = 31-33). Thus, the expression of this gene can be used to distinguish whether this tissue is of fetal or adult origin. The relatively overexpression of the gene in these fetal tissues suggests that the gene product increases the growth or development of these organs in the fetus and can also act as a regenerative capacity in adults. Therefore, therapeutic modulation of the protein encoded by this gene may be useful in the treatment of diseases affecting these organs.

  Among metabolic tissues, this gene is highly expressed in pancreas, kidney, fetal liver, and adipocytes. It is also expressed at moderate levels in the pituitary gland, thyroid, heart, fetus and adult skeletal muscle, with low but significant expression in the liver and fetal heart.

  Panel 2.2 Summary: Ag3857 The highest expression is found in kidney cancer (CT = 28.2). In addition, this gene is more highly expressed in kidney cancer than in the corresponding normal adjacent tissue. Thus, the expression of this gene could be used as a marker for this cancer. Furthermore, therapeutic modulation of the expression or function of the gene product may be useful for the treatment of kidney cancer.

  Panel 4.1D Overview: Ag3857 The highest expression of this gene is detected in IL-4 treated dermal fibroblasts (CT = 25.3). In addition, high levels of expression are also observed in a group of samples derived from dermal fibroblasts. Thus, the expression of this gene can be used as a marker for this cell. In addition, therapeutic modulation of the expression or function of this gene may be useful for the treatment of skin disorders such as psoriasis.

  Panel 5 Islet Summary: Ag3857 (autotaxin-t) is expressed in human islets (CT = 30). Autotaxin has been found to hydrolyze the type I phosphodiesterase substrate p-nitrophenylthymidine-5'-monophosphate (J Biol Chem 1994 Dec 2: 269 (48): 30479-84), phosphodiesterase (PDE) It is. In a CuraGen GeneCalling study, it was found that the rat ortholog (PDE1) was down-regulated in a good vs. low secretory cell line. Thus, this antagonist to autotaxin could improve insulin secretion in type 2 diabetes.

  Panel 5D Summary: Ag3857 The highest expression of this gene is found in adipocytes (CT = 28.7). Medium levels of expression are found in other metabolic tissues of this panel, such as skeletal muscle. Overall, these results are consistent with those of panel 5I. See that panel for further discussion of the use of this gene in metabolic diseases.

BV. CG93541-02: Autotaxin-like The expression of the gene CG935541-02 was evaluated using the primer-probe set Ag6912 described in Table BVA. The results of RTQ-PCR execution are shown in Table BVB.

Continuation of Table BVB

  General_Screening_Panel_v1.6 Summary: The highest expression of the Ag6912 CG93541-02 gene is found in the spinal cord (CT = 30.3). In addition, moderate to low levels of expression are found in the tonsils, cerebellum, cerebral cortex, thalamus, substantia nigra and whole brain. This gene encodes a protein homologous to autotaxin. Autotaxin variants have been found to be concentrated in the brain and spinal cord, and their expression is found in oligodendrocytes. This autotaxin expression is highest in adults. (Fuss, B. J Neurosci 1997 Dec 1:17 (23): 9095-10). Based on the homology of this gene to autotaxin and its expression profile in this panel, expression of this gene can be used as a marker for neuronal tissue. Also, therapeutic modulation of the expression or function of this gene may be useful in the treatment of neurological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia, multiple sclerosis, stroke and epilepsy.

Among metabolic tissues, this gene is expressed at low but significant levels in adipocytes, fetal liver, heart, adrenal gland and pituitary. This expression suggests that this gene product has a role in normal neuroendocrine and metabolic functions, and that unregulated expression of this gene may contribute to neuroendocrine disorders or metabolic diseases such as obesity and diabetes .
Medium level expression is also observed in a group of samples from melanoma cell lines. Thus, therapeutic modulation of the expression or function of this gene may be effective in the treatment of melanoma.

BW. CG95872-02: Aurora-related kinase 2
The expression of the gene CG95872-02 was evaluated using the primer-probe set Ag6958 described in Table BWA. The results of RTQ-PCR execution are shown in Table BWB.

Continuation of Table BWB

  General_Screening_Panel_v1.6 Summary: The highest expression of the Ag6958 CG95872-02 gene is found in colon cancer cell lines (CT = 25.7). Overall, the expression of this gene appears to be associated with cancer cell lines, with high to medium levels of expression seen in all cell lines on this panel. Furthermore, moderate levels of expression are found in all fetal tissues on this panel, while expression in the corresponding adult tissues is low / undetectable. This gene encodes a protein with homology to Aurora kinases involved in chromosome segregation and cytokinesis. Members of this family have been shown to be expressed with high mitotic index in cells such as fetal liver and cancer cell lines (Bischoff J.R. Trends in Cell Biology 1999, 9: 454-459). Furthermore, members of this Aurora kinase family have been shown to bind to the SH3 domain of RasGAP, which has anti-apoptotic effects, in the control of cell proliferation. (Gigoux V. J Biol Chem 2002 Apr 25). Based on the homology of this gene to Aurora kinase and the preferential expression in proliferating tissues, the expression of this gene could be used as a cancer marker. Furthermore, therapeutic modulation of the expression or function of this protein may be effective in the treatment of cancer.

Example D: Identification of single nucleotide polymorphisms (SNPs) in NOVX nucleic acid sequences Variant sequences are also included in this application. Variant sequences can include single nucleotide polymorphisms (SNPs). SNP is referred to in some examples as “cSNP” and refers to the nucleotide sequence containing the SNP originating as cDNA. SNPs can occur in several ways. For example, the SNP can be for replacement of one nucleotide of a polymorphic site with another. Such substitutions can be transitions or transversions. SNPs can also result from the deletion or insertion of nucleotides from the reference allele. In this case, the polymorphic site is a site where one allele has a gap for a particular nucleotide of another allele. A SNP present in the absence of a gene can result in a change in the amino acid encoded by the gene at the position of the SNP. Intragenic SNPs can also be silent, with codons containing SNPs encoding the same amino acid as a result of the degeneracy of the genetic code. SNPs present outside the gene region or in introns within the gene do not change any amino acid sequence of the protein, but the regulation of the expression pattern may be altered. Examples include transient expression, physiological response regulation, cell type expression regulation, intensity of expression, and transcriptional messenger stability.

  The SeqCalling assembly produced by the exon linking process was selected and stretched using the following criteria. Genomic clones with regions with 98% identity to all or part of the original or extended sequences were identified by BLASTN search using related sequences to the query human genome database. The resulting genomic clones were selected for further analysis, since this identity points out that these clones contain the genomic locus for the SeqCallng assembly. These sequences were analyzed for putative coding regions and for similarity to known DNA and protein sequences. Programs used for these analyzes include Grail, Genscan, BLAST, HMMER, FASTA, Hybrid, and other related programs.

  Some additional genomic regions were also identified because we selected SeqCalling assembly maps to those regions. Such SeqCalling sequences overlapped with regions defined by homology or exon prediction. These can also be included because the position of the fragment was in the vicinity of the genomic region identified by similarity or exon prediction originally included in the predicted sequence. The sequences so identified could be assembled manually and then extended using one or more additional sequences taken from the CuraGen Corporation human SeqCalling database. SeqCalling fragments suitable for inclusion were identified by the CuraTools® program SeqExtend or by identifying SeqCalling fragment mapping to the appropriate regions of the analyzed genomic clone.

  The regions defined by the above technique are then manually integrated and for obvious discrepancies that can arise, for example, from miscored bases in the original fragment or from discrepancies between regions of predicted exon binding, EST position and sequence similarity Corrected and derived the final sequence disclosed herein. When necessary, the process for identifying and analyzing SwqCalling assemblies and genomic clones was repeated to derive full-length sequences (Alderborn et al., Determination of Single nucleotide Polymorphisms by Real-time Pyrophosphate DNA Sequencing.Genome Research. 10 (8) 1249-1265, 2000).

  Variants are reported individually, but any combination of all or a selected subset of variants is also encompassed as a contemplated NOVX embodiment of the invention.

Results NOV2a SNP data 7 NOV2a polymorphic variants were identified. Shown in Table 2S.

NOV3a SNP data Two polymorphic variants of NOV3a were identified. Shown in Table 3S.

NOV8a SNP data One polymorphic variant of NOV8a was identified. Shown in Table 8S.

NOV11a SNP data One polymorphic variant of NOV11a was identified. Shown in Table 11S.

NOV15a SNP data Five polymorphic variants of NOV15a were identified. Shown in Table 15S.

NOV16a SNP data One polymorphic variant of NOV16a was identified. Shown in Table 16S.

NOV17a SNP data 18 polymorphic variants of NOV17a were identified. Shown in Table 17S.

NOV20a SNP data 15 polymorphic variants of NOV20a were identified. Shown in Table 20S.

NOV21a SNP data Seven polymorphic variants of NOV21a were identified. Shown in Table 21S.

NOV23a SNP data One polymorphic variant of NOV23a was identified. Shown in Table 23S.

NOV25a SNP data One polymorphic variant of NOV25a was identified. Shown in Table 25S.

NOV26a SNP data Two polymorphic variants of NOV26a were identified. Shown in Table 26S. .

NOV27a SNP data One polymorphic variant of NOV27a was identified. Shown in Table 27S.

NOV30a SNP data One polymorphic variant of NOV30a was identified. Shown in Table 30S.

NOV31a SNP data Five polymorphic variants of NOV31a were identified. Shown in Table 31S.

NOV38a SNP data Nine polymorphic variants of NOV38a were identified. Shown in Table 38S.

NOV39a SNP data Four polymorphic variants of NOV39a were identified. Shown in Table 39S.

NOV40a SNP data One polymorphic variant of NOV40a was identified. Shown in Table 40S.

NOV42a SNP data One polymorphic variant of NOV42a was identified. Shown in Table 42S.

NOV46a SNP data Five polymorphic variants of NOV46a were identified. Shown in Table 46S.

NOV50a SNP data One polymorphic variant of NOV50a was identified. Shown in Table 50S.

NOV51a SNP Data One polymorphic variant of NOV51a was identified. Shown in Table 51S.

NOV57a SNP data One polymorphic variant of NOV57a was identified. Shown in Table 57S.

NOV66a SNP data Seven polymorphic variants of NOV66a were identified. Shown in Table 66S.

NOV70a SNP data Four polymorphic variants of NOV70a were identified. Shown in Table 70S.

NOV71a SNP data Five polymorphic variants of NOV71a were identified. Shown in Table 71S.

Example E.1. NOV45—Method of Use The present invention is based in part on the identification of biological macromolecules that are differentially regulated in disease states, diseases, or abnormal conditions or conditions, and / or cDNA libraries or single matrices Based on novel associations of proteins and polypeptides identified in yeast two-hybrid screening with reactions and the nucleic acids that encode them. Such lesions or diseases include endocrine diseases, cancer, various tumors and abnormal growth, inflammatory diseases, central nervous system diseases, and metabolic diseases including those associated with similar abnormal conditions or conditions. Significant metabolic diseases to which biological macromolecules are related include obesity and diabetes, particularly obesity and type 2 diabetes. Obesity is thought to predispose to type 2 diabetes. In a very important embodiment of the invention, the biological macromolecules are related in that the lesions and conditions are proteins and polypeptides, in which case the invention also relates to the nucleic acids that encode them. Related. Methods that can be utilized to identify related biological macromolecules include any method that detects differential expression of nucleic acids encoding proteins and polypeptides associated with disease, as well as the respective proteins and polypeptides. It includes a method for detecting the peptide itself. An important method that has been utilized by the present invention is the GeneCalling® technology disclosed in US Pat. No. 5,871,697 and US Serial No. 09 / 417,386, filed Oct. 13, 1999, respectively. And SeqCalling® technology, each of which is incorporated herein by reference in its entirety. GeneCalling® is also disclosed in Shimkets, et al., Nature Biotechnology 17: 198-803 (1999).

  The present invention provides polypeptides and nucleotides that encode those identified as having a novel association with a disease or lesion, or abnormal condition or condition in a mammal. In the present invention, nucleic acid sequences and their encoded polypeptides are included. The sequence is generally associated as “obesity and / or diabetes nucleic acid” or “obesity and / or diabetes polynucleotide” and the counterpart encoding the polypeptide is “obesity and / or diabetes polypeptide” or “obesity and And / or “diabetic protein”. For example, an obesity and / or diabetes nucleic acid according to the present invention is a nucleic acid comprising an obesity and / or diabetes nucleic acid, and an obesity and / or diabetes polypeptide associated with the present invention comprises the amino acid sequence of an obesity and / or diabetes polypeptide. It is a polypeptide. Despite being indicated, “obesity and / or diabetes” is meant to relate to any sequence having a novel association disclosed herein.

  The present invention includes naturally occurring polypeptides, precursor forms or proproteins, or mature forms of polypeptides or proproteins that are relevant as targets for therapeutic reagents in the treatment of various diseases, pathologies, abnormal states and abnormal conditions. Identify protein and polypeptide sets. The target can be utilized in any of a variety of screening methods to identify candidate therapeutic reagents that interact with the target and thereby exert the desired or favorable effect. Therapeutic reagent candidates are identified in important embodiments of the invention by screening a large collection of substances or mixtures. Such a collection may include a library of combinations of substances or mixtures, in which at least one subset of substances or mixtures, each member is related by a simple structural variation based on a specific limit structure or basic chemical structure, respectively. Variations include, without limitation, changes in the basic framework length or identity of the bonded atoms; changes in numbers, cyclic structures, bridge structures, alicyclics, aromatic ring compositions and qualities; And may include ongoing frameworks or substituted atoms or groups of substituents attached at specific positions to the basic framework or ring structure, bridge structure, alicyclic ring, aromatic ring of bonded atoms.

  The present invention discloses novel associations of proteins and polypeptides and nucleic acids that encode those encoded in yeast two-hybrid screens using cDNA libraries or one-by-one matrix reactions. Proteins and related proteins similar to those encoded by cDNA and / or genomic DNA have been identified in some cases by CuraGen Corporation.

  In the present invention, protein interactions may include interaction of a protein fragment having a full-length protein, a protein fragment having another protein fragment, or a full-length protein having each. The protein interactions disclosed in the present invention also represent functionally significant significant discoveries for specific diseases or conditions. Here, a new protein is seen as a component of a known pathway, a known protein is seen as a component of a novel pathway, or a known protein is seen as a component of a novel pathway.

  Polypeptides or proteins utilized as targets in the screening methods disclosed herein include naturally occurring polypeptide or precursor forms or proprotein products. Naturally occurring polypeptides, precursors or proproteins include, for example, the full-length gene product encoded by the corresponding gene. Naturally occurring polypeptides also include polypeptides, precursors or proproteins encoded by the open reading frames disclosed herein. A “mature” form of a polypeptide or protein occurs as a result of one or more naturally occurring processing steps that can occur in cells, including host cells. The processing step occurs, for example, as a gene product resulting from cleavage of the amino terminal methionine residue by the start codon of the open reading frame, or proteolytic cleavage of the signal peptide or leader sequence. Thus, the mature form resulting from a precursor polypeptide or protein having residues 1 to N, where residue 1 is the N-terminal methionine, has residue 2 through N residues. Alternatively, the mature form resulting from a precursor polypeptide or protein having residues 1 to N where the amino terminal signal sequence from residue 1 to residue M is cleaved is here the amino terminus from residue 1 to residue M The signal sequence is cleaved to include residues from residue M + 1 to residue N. A “mature” form of a polypeptide or protein can also result from non-proteolytic post-translocational modifications. Such non-proteolytic processes include, for example, glycosylation, myristylation or phosphorylation. In general, a mature polypeptide or mature protein can result from only one manipulation of these processes, or any combination thereof.

  As used herein, “identical” residues correspond to those residues in a two-sequence comparison in which the same nucleotide base or amino acid residue is the same residue in an alignment of the two sequences. Residues are alternatively described as “similar” or “positive” when a comparison between two sequences in an alignment is the same amino acid or a conserved amino acid as defined below, in the same position in the comparison.

  As used herein, “chemical composition” relates to a composition comprising at least one composition synthesized or extracted from a natural source. A chemical composition can be the product of a defined synthetic method. Such synthesized compositions have the properties defined in terms of molecular formulas, molecular structures related to the association of atoms attached to each, physical properties such as electropherographic or spectroscopic characterization, etc. herein. Understood in the book. Compositions extracted from natural sources provide a representation of defined properties, including molecular formulas, molecular structures related to the association of atoms attached to each, physical properties such as electropherography or spectroscopic characterization, etc. It is usefully analyzed by chemical and physical methods.

  As used herein, a “therapeutic reagent candidate” is a chemical composition comprising at least one substance that appears to bind to a target biopolymer. In important embodiments of the invention, the target biopolymer is a lipid such as a protein, polypeptide, nucleic acid, polysaccharide or proteoglycan, or complex lipid. A method for identifying a composition that binds to a target effectively eliminates a composition that has little or no binding affinity, and the potential for the identified chemical composition to have advantageous therapeutic applications. Increase. In the case where a “therapeutic reagent candidate” is a mixture of more than one chemical composition, subsequent screening methods are performed to identify specific substances that bind to the composition in the mixture and identify them as therapeutic reagent candidates. The

  As used herein, a “medicament” is a therapeutic reagent that uses a model of a disease state or pathology to identify candidates that have a desired therapeutic or beneficial therapeutic effect on an association with the disease or pathology. Provided by screening candidates. A candidate that successfully provides such an effect has been named herein a pharmaceutical product. Non-limiting examples of model systems used in such sorting include specific cell lines, cultured cells, tissue specimens, whole tissues, organ specimens, intact organs, and non-human mammals. Sorting utilizing at least one system, preferably more than one system, can be utilized to identify pharmaceuticals. The identified pharmaceutical can be explored in further studies using human subjects.

  The following stages identify design studies and for repeat ion channel-encoded NOV45 proteins, and any variants thereof, and their stability as diagnostic markers, targets for antibody therapy, targets for small molecules against obesity and diabetes Describe the technology used to demonstrate.

Four repeat ion channel CG105597-01
Four-repeat ion channels are novel calcium channels that are exclusively expressed in adipocyte differentiation and in the brain.

  Pharmacological interference with fat storage has been recognized as a powerful approach to anti-obesity treatment. Peroxisome proliferator-activated receptor γ (PPARγ) is a nuclear hormone receptor that has been found to be sufficient and necessary for the differentiation of new adipocytes. Activation of PPARγ leads to an increase in the circulation of free fatty acids (FFA) and improves insulin sensitivity in the periphery. Their entire thiazolidinedione, the various PPARγ ligands, is a marker for the treatment of type 2 diabetes. However, these compositions do not act against obesity and, on the other hand, promote weight gain. Inhibitors of the PPARγ pathway are contemplated for the treatment of obesity because they are intended to inhibit the recruitment of new adipocytes.

Intracellular calcium [Ca ²⁺ ] _i has been implicated in the regulation of adipogenesis. An increase in [Ca ²⁺ ] _i plays a biphasic regulatory role in human adipocyte differentiation. It works to inhibit the early stages of differentiation while promoting the late stages of differentiation and lipid loading. An increase in [Ca ²⁺ ] _i has resulted in a marked increase in the expression of PPARγ. Increased expression of PPARγ directly affects and induces adipocyte differentiation by directly acting on and inducing late differentiation gene expression such as aP2, stearoyl CoA desaturase (SCD-1), phosphoenolpyruvate carboxykinase (PEPCK) and FAS Can be promoted. On the other hand, in late differentiation, preadipocytes are CCAAT / enhancer binding proteins β and δ (C / EBPβ and C / EBPΔ), PPARγ, and sterol regulatory element binding protein 1 / adipocyte determination and differentiation factor 1 (SREBP-1). Commit by terminal differentiation after expression of some clinical differentiation transcription factors such as / ADD-1). [Ca ²⁺ ] _i is synthesized with these transcription factors and can promote the differentiation program by stimulating late differentiation gene expression. In view of this, [Ca ²⁺ ] _i behaves as a lipogenic and anti-lipolytic signaling factor in the control of adipocyte metabolism in both rodents and human adipocytes. An increase in [Ca ²⁺ ] _i has been seen to be a key enzyme in new adipogenesis, thereby stimulating FAS expression and activity that promotes storage of triglycerides. In addition, an increase in [Ca ²⁺ ] _i has been seen to inhibit basal lipolysis and agents that stimulate lipolysis in early cultured human adipocytes. These effects can be completely blocked by calcium channel antagonists. Thus, an increase in [Ca ²⁺ ] _i exerts and regulates regulation in adipogenesis and lipolysis, simultaneously stimulating the first half and suppressing the second half, thereby leading to lipid loading and adipocyte hypertrophy It seems to promote the accumulation of triglycerides in adipocytes. Therefore, the adipogenesis and antilipolytic action of [Ca ²⁺ ] _i coupled with increased expression of differentiation transcription factors such as PPARγ may contribute to the stimulatory action of [Ca ²⁺ ] _i in later stages of differentiation. Thus, the reduction of intracellular calcium in the late stages of adipose differentiation by antagonists of the four repeat ion channel may be effective in the treatment of obesity. Rosen et al. 2002 Genes Dev 16 (1): 22-26; Ren et al., 2002 Genes Dev 116 (1): 27-32; Lazar 2002 Genes Dev. 16 (1): 1-5; Shi et al ., 2000 Physiol Genomics 3 (2): 75-82; Spiegelman and Flier 1996 Cell 87: 377389 in general.

NOV45 expression and clustering analysis CG105597-01 (which is of course also involved as NOV45) was identified based on up-regulation in adipocyte differentiation in quantitative expression analysis of clones in various cells and tissues. CG105597-01 was up-regulated 3-5 fold in midway differentiated adipocytes when compared to undifferentiated mesenchymal cells. The expression of CG105597-01 was lower in fully differentiated adipocytes.

  In cluster formation analysis, CG105597-01 shows similar expression characteristics to PPARγ, indicating that CG105597-01 and PPARγ can function in the same pathway that regulates adipocyte differentiation.

  Cluster formation analysis was performed by comparing the metabolic properties of all CuraGenRTQ-PCR probes with the expression properties of metabolic panel 5I with that of CG105597-01 probe Ag4292. The results are shown in Table E1.

Table E1. Query: CG105597-01

  The expression characteristics of CG105597-01 are very similar to that of PPARγ, indicating that CG105597-01 can work in the PPARγ pathway in regulating adipocyte differentiation.

Intracellular pathways In certain embodiments, the four-repeat ion channel protein NOV45 is involved in adipocyte differentiation. Pharmacological interference with fat storage has been recognized as a powerful approach to anti-obesity treatment. Peroxisome proliferator-activated receptor γ (PPARγ) has been considered sufficient and necessary for the development of new adipocytes. Activation of PPARγ leads to a circulating increase in free fatty acids (FFA) and an improvement in peripheral insulin sensitivity. Various PPARγ ligands, all of which are thiazolidinediones, are on the market for the treatment of type II diabetes. However, these mixtures have no effect on obesity and, on the other hand, can promote weight gain. Inhibition of the PPARγ pathway is intended to inhibit the introduction of new adipocytes and can be considered for the treatment of obesity. Intracellular calcium [Ca ²⁺ ] _i has been implicated in the control of adipogenesis. An increase in [Ca ²⁺ ] _i functions to inhibit the early stages of differentiation, while promoting the late stages of differentiation and lipid loading and exerts a biphasic regulatory role in human adipocyte differentiation.

Pathways Associated with Obesity and / or Diabetes Etiology and Cause The following illustration shows methods of alteration in expression of human NOV45 for repeat ion channels and gene product function methods associated with the pathogenesis and cause of obesity and / or diabetes. The schematic captures the characteristic findings of these discovery studies in combination with those reported in the literature. The result of inhibition of human four-repeat ion channel action is the inhibition of new adipocyte differentiation and new adipocyte formation, a phenomenon contributing to weight gain and obesity. The adipogenic pathway is shown in Table E2.

Table E2. Intracellular pathway of adipogenesis

The rationale for use as a diagnostic / and target for small molecule drugs and antibody therapy The following is a summary of findings from discovery studies, supplementary studies that also incorporate knowledge in the scientific literature, and assays. Overall, the data indicate that inhibitors / antagonists of human for repeat ion channels may be beneficial in the treatment of obesity and / or diabetes.

  Four-repeat ion channels are upregulated in adipocyte differentiation. The expression characteristics of the for repeat ion channel are closely similar to that of PPARγ. PPARγ is sufficient and necessary for adipogenesis. For-repeat ion channels may work downstream of PPARγ in promoting adipogenesis. Inhibitors of the four repeat ion channel can consequently delay the inhibition of adipocyte differentiation, de novo adipocyte synthesis and weight loss. Therefore, novel antagonists of four repeat ion channels may be beneficial in the treatment of obesity.

Antibodies The present invention further includes antibodies and antibody fragments such as Fab, (Fab) ₂ or single chain _Fv constructs that immunospecifically bind to any protein of the present invention. In the present invention, it has a high binding affinity for any protein of the present invention comprising such peptides and polypeptides fused to any carrier particle (or biologically expressed on the carrier surface) such as bacteriophage particles. Also included are peptides and polypeptides comprising sequences.

Use of the Compositions of the Invention Expression information, subcellular localization, protein interaction complexes and the protein and nucleic acid map positions disclosed herein, which are similar information for a protein, are expressed by the protein as repeats. It shows that it can have important structural and / or physiological functional characteristics of the ion channel family. The nucleic acids and proteins of the invention are therefore useful in potential diagnostic and therapeutic applications and as research tools. These include acting as specific or selective nucleic acid or protein diagnostic and / or prognostic markers, where the presence or amount of the nucleic acid or protein can be assessed. These also include potential therapeutic applications such as: (i) protein therapy, (ii) small molecule drug targets, (iii) antibody targets (therapeutic, diagnostic, drug targets / cytotoxic antibodies), ( iv) nucleic acids useful in gene therapy (gene transfer / gene cleavage), (v) reagents that promote tissue regeneration in vitro and in vivo, and (vi) biological defense weapons.

  The nucleic acids and proteins of the present invention have application in the diagnosis and / or treatment of various diseases and conditions. For example, the compositions of the invention will have efficacy for the treatment of patients suffering from obesity and / or diabetes.

  These substances are further useful in the production of antibodies that immunospecifically bind to the substances of the invention for use in diagnostic and / or therapeutic methods.

Example F.1. Utility of NOV70 in tumorigenesis SAGE analysis, intracellular and in vivo confirmation assays, knockdown cell confirmation were performed as disclosed below.

  Potential role of CG59693-01 in tumorigenesis: There is a subset of lung tumors, particularly non-small cell lung tumors, associated with significant exacerbation results. This phenotype is strongly associated with the ability of these tumors to be resistant to chemotherapy. As indicated above, this gene is overexpressed in a subset of lung tumors (see TaqMan data and Hsu et al Cancer Res 2001 Mar 15; 61 (6): 2727-31). Antisense data indicates a decrease in the activity of this enzyme that reduces drug resistance levels. This decrease is associated with improved clinical outcome.

  Effect of therapeutic target of CG59693-01: The therapeutic target of the enzymatic activity of the protein encoded by CG59693-01 with a small molecule inhibitor reduces the tolerance limit to chemotherapy in lung cancer, particularly non-small cell lung tumors Expected to be abolished.

Continuous gene expression analysis of CG59693-01 in various cells and tissues A continuous analysis of gene expression (SAGE) using a short polynucleotide sequence, typically a SAGE tag of about 20 nucleotides or less, occurs at a position in the messenger RNA. SAGE tags can be used to identify corresponding transcripts and transcribed genotypes. SAGE analysis begins by providing complementary deoxyribonucleic acid (cDNA) from a tumor cell line-derived library. The cDNA can bind to the primer site. The sequence tag is then generated, for example, using suitable primers to synthesize DNA. By measuring differences in these labeled tags between cDNA libraries, sequences that are aberrantly expressed in tumor cell lines can be identified. SAGE was performed with CG59693-01 nucleic acid and is shown in Table F1.

Table F1. SAGE data

EST analysis data:
Positive signals were obtained in fetal liver / spleen; liver; gallbladder; infant brain; colon; islet; brain; fetal lung;

Suggested intracellular and in vivo validation assay CG59693-01 was further analyzed in the following assay. Two intracellular assays were performed and tested for chemosensitivity. In the first assay, the knockdown of CG59693-01 expression in NCI-H460 (positive cell line) was analyzed. In the second assay, CG59693-01 was stably transfected into NCI-H460 and NCI-N417 (null cell line) and analyzed. Further data were obtained in an in vivo assay utilizing a drug resistant animal model. Knock-in cell lines were NCI-H1299, NCI-H157 and NCI-N417. The knockout cell line was NCI-H460.

Knockdown cell confirmation:
All oligonucleotides are mixed backbone oligonucleotides containing a modified phosphorothione segment at the 5 'and 3' ends and an RNA oligoribonucleotide segment located in the middle. Oligonucleotides were synthesized by Midland, Inc. All oligonucleotides were desalted and gel purified. The purity of the oligonucleotide was confirmed by mass spectrometry.

  Untransfected cells, NCI-H460 cells transfected with a mix of oligonucleotides and NCI-H460 cells transfected with CG59693-01 antisense oligonucleotide were treated with paclitaxel for 48 hours. Cell survival was measured as the ratio of the absorbance of cells treated with drug to untreated cells.

  The CG59693-01 knockdown assay had little effect on cell growth in NCI-H460 cells after 48 hours. For example, Hsu et al., Cancer Res 2001 Mar 15; 61 (6): 2727-31; Matsuura et al., Biochem J 1997 Apr 1; 323 (Pt 1): 61-4; Deng et al., J Biol See also Chem 2002 Feb 12; (electronic publication ahead of print); Miyabe Y et al., Yakugaku Zasshi 1997 Mar; 117 (3): 167-77.

NOV70e knockdown assay

NOV70e knockdown assay-48 hours paclitaxel treatment

NOV70e knockdown assay-cell survival

NOV70e knockdown assay-72 hours paclitaxel treatment

NOV70e knockdown assay-cell survival

(Other embodiments)
Although particular embodiments are disclosed herein in detail, this is done for purposes of illustration only and is not intended to limit the scope of the following appended claims. In particular, it is contemplated by the inventor that various substitutions, changes and modifications may be made to the invention without departing from the spirit and scope of the invention as defined by the claims. The choice of nucleic acid starting material, desired clone, or library type is believed to be a routine matter of ordinary skill in the art having knowledge of the embodiments described herein. Other aspects, advantages, and modifications are considered to be within the scope of the following claims. The claims presented are representative of the invention disclosed herein. In addition, non-claimed inventions are also contemplated. Applicant retains the right to pursue such inventions in the following claims.

Claims (45)

  An isolated polypeptide comprising the mature form of the amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539.   An isolated polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539.   An isolated polypeptide comprising an amino acid sequence that is at least 95% identical to an amino acid selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 239.   An isolated polypeptide, wherein the polypeptide comprises an amino acid sequence comprising one or more conservative substitutions in an amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539 .   The polypeptide of claim 1, wherein the polypeptide is naturally occurring.   A composition comprising the polypeptide of claim 1 and a carrier.   A kit comprising the composition of claim 6 in one or more containers.   A therapeutic use in the manufacture of a medicament for treating a syndrome associated with a human disease, wherein the disease is selected from lesions associated with the polypeptide of claim 1 and comprises a polypeptide comprising the polypeptide of claim 1 Use in. A method for determining the presence or amount of a polypeptide of claim 1 in a sample comprising:
(A) preparing the sample;
(B) introducing the sample into an antibody that immunospecifically binds to the polypeptide; and (c) measuring the presence or amount of the antibody that binds to the polypeptide;
Including
A method thereby measuring the presence or amount of a polypeptide in the sample. A method for determining the presence or predisposition of a disease associated with an altered level of expression of a polypeptide of claim 1 in a first mammalian subject comprising:
a) measuring the level of expression of the polypeptide in the sample from the first mammalian subject; and b) not having the disease or predisposition to expression of the polypeptide in the sample of step (a) Comparing the expression of the polypeptide present in a control sample from a second mammalian subject known from
The method wherein a change in the expression level of the polypeptide in the first subject when compared to a control sample indicates the presence or predisposition of the disease. A method for identifying a reagent that binds to the polypeptide of claim 1 comprising:
(A) introducing the polypeptide into the reagent; and (b) determining whether the reagent binds to the polypeptide;
Including methods.   12. The method of claim 11, wherein the reagent is a cellular receptor or a downstream effector. A method of identifying a potential therapeutic agent for use in the treatment of a lesion, wherein the lesion is associated with aberrant expression or aberrant physiological interactions of the polypeptide of claim 1;
(A) preparing a cell that expresses the polypeptide of claim 1 and has a property or function attributable to the polypeptide;
(B) contacting the cell with a composition comprising a candidate substance; and (c) determining whether the substance alters a property or function attributable to the polypeptide;
Thereby, if a change observed in the presence of a substance is not observed when the cell is contacted with the composition in the absence of the substance, the substance is identified as a potential therapeutic agent. A method of screening for modulators of the activity, potential activity, or predisposition of a lesion associated with the polypeptide of claim 1 comprising:
(A) administering a test compound to a test animal having an increased risk of a lesion associated with the polypeptide of claim 1, provided that the test animal receives the polypeptide of claim 1; Expressed recombinantly;
(B) measuring the activity of the polypeptide in the subject animal after administration of the compound of step (a); and (c) administering the polypeptide to determine the activity of the polypeptide in the subject animal. Comparing the activity of the polypeptide in an untreated control animal, provided that the change in activity of the polypeptide in the test animal compared to the control animal is determined by the test compound according to claim 1. To indicate a potential activity or predisposing modulator of a lesion associated with the peptide;
Including the method.   The test animal expresses a test protein transgene or is a recombinant test animal that expresses the transgene under the control of a promoter at an increased level compared to a wild-type test animal, wherein 15. The method of claim 14, wherein the promoter is not the native gene promoter of the transgene.   A method of modulating the activity of a polypeptide according to claim 1, wherein the polypeptide of claim 1 is expressed together with a compound that binds to the polypeptide in an amount sufficient to modulate the activity of the polypeptide. Contacting the cell sample.   A method of treating or preventing a lesion associated with the polypeptide of claim 1, wherein said subject is desired in an amount sufficient to treat or prevent a lesion in the subject. A method comprising administering the polypeptide according to 1.   The method of claim 17, wherein the subject is a human.   A method of treating a morbidity in a mammal, comprising administering the polypeptide to the mammal in an amount sufficient to alleviate the morbidity, wherein the polypeptide comprises SEQ ID NO: 2n, where n is 1 to 129) and a biologically active fragment thereof having an amino acid sequence at least 95% identical to a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 539.   An isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538.   21. The nucleic acid molecule of claim 20, wherein the nucleic acid molecule is naturally occurring.   A nucleic acid molecule wherein the nucleic acid molecule differs from a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 by one nucleotide.   An isolated nucleic acid molecule which encodes a mature form of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 2n (n is an integer from 1 to 129) and SEQ ID NO: 539.   An isolated nucleic acid molecule comprising a nucleic acid selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538.   The nucleic acid molecule is under stringent conditions to a nucleotide sequence selected from the group consisting of SEQ ID NO: 2n-1 (n is an integer from 1 to 129) and SEQ ID NO: 538 or a complementary sequence of the nucleotide sequence 21. The nucleic acid molecule of claim 20, which hybridizes with.   A vector comprising the nucleic acid molecule of claim 20.   27. The vector of claim 26, further comprising a promoter operably linked to the nucleic acid molecule.   A cell comprising the vector of claim 26.   An antibody that immunospecifically binds to the polypeptide of claim 1.   30. The antibody of claim 29, which is a monoclonal antibody.   30. The method of claim 29, wherein the method is a humanized antibody. A method for measuring the presence or amount of a nucleic acid molecule according to claim 20 in a sample comprising:
(A) preparing the sample;
(B) introducing the sample into a probe that binds to the nucleic acid molecule; and (c) measuring the presence or amount of the probe that binds to the nucleic acid molecule;
Including
Thereby measuring the presence or amount of nucleic acid molecules in the sample.   35. The method of claim 32, wherein the presence or amount of the nucleic acid molecule is used as a marker for a cell or tissue type.   34. The method of claim 33, wherein the cell or tissue type is cancerous. A method for determining the presence or predisposition of a disease associated with an altered level of expression of a nucleic acid molecule of claim 20 in a first mammalian subject comprising:
a) measuring the level of expression of the nucleic acid in the sample from the first mammalian subject; and b) the expression level of the nucleic acid in the sample of step (a) is known not to be diseased or predisposed Comparing the level of expression of the nucleic acid present in a control sample from a second mammalian subject;
Including
The method wherein a change in the level of expression of the nucleic acid in the first subject when compared to the control material indicates the presence or predisposition of the disease.   A method for producing the polypeptide according to claim 1, wherein the cell is cultured under conditions that induce expression of the polypeptide, provided that the cell is SEQ ID NO: 2n-1 (n is an integer from 1 to 129). And a vector comprising an isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 538.   40. The method of claim 36, wherein the cell is a bacterial cell.   38. The method of claim 36, wherein the cell is an insect cell.   37. The method of claim 36, wherein the cell is a yeast cell.   38. The method of claim 36, wherein the cell is a mammalian cell.   A method for producing the polypeptide according to claim 2, wherein the cell is cultured under conditions that induce expression of the polypeptide, provided that the cell is SEQ ID NO: 2n-1 (n is an integer of 1 to 129). And a vector comprising an isolated nucleic acid molecule comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO: 538.   42. The method of claim 41, wherein the cell is a bacterial cell.   42. The method of claim 41, wherein the cell is an insect cell.   42. The method of claim 41, wherein the cell is a yeast cell.   42. The method of claim 41, wherein the cell is a mammalian cell.