TW201143782A - Treatment of LIM homeobox 2 (LHX2) related diseases by inhibition of natural antisense transcript to LHX2 - Google Patents

Abstract

The present invention relates to antisense oligonucleotides that modulate the expression of and/or function of LIM homeobox 2 (LHX2), in particular, by targeting natural antisense polynucleotides of LIM homeobox 2 (LHX2). The invention also relates to the identification of these antisense oligonucleotides and their use in treating diseases and disorders associated with the expression of LHX2.

Description

201143782 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION Embodiments of the invention include oligonucleotides and related molecules that modulate the performance and/or function of LHX2. The present application claims priority to U.S. Provisional Patent Application Serial No. 61/346,284, filed on Jan. [Prior Art] O DNA-RNA and RNA-RNA hybridization are important for many nucleic acid functional aspects including DNA replication, transcription, and translation. Hybridization is also critical for the detection of specific nucleic acids or the various techniques that alter their performance. For example, antisense nucleotides disrupt gene expression by hybridizing to a target RNA, thereby interfering with RNA splicing, transcription, translation, and replication. Antisense DNA has the additional property that DNA-RNA hybrids can be used as receptors for ribonuclease digestion, which is present in most cell types. The antisense molecule can be delivered to a cell, such as in the case of an oligodeoxynucleotide (ODN), or it can be expressed as an RNA molecule from an endogenous 〇 gene. The FDA recently approved the antisense drug VITRAVENETM (for the treatment of cytomegalovirus retinitis), indicating that this antisense drug has therapeutic utility. BRIEF DESCRIPTION OF THE DRAWINGS The Summary is provided to introduce a summary of the invention This Summary is submitted with the understanding that it is not to be construed as limiting or limiting the scope or meaning of the scope of the application. In one embodiment, the invention provides a method of inhibiting the action of a natural antisense transcript 156342.doc 201143782 by upregulating an antisense oligonucleotide by targeting any region of a natural antisense transcript The righteous gene is reached. It is also encompassed herein that inhibition of natural antisense transcripts by siRNA, ribozymes and small molecules is considered to be within the scope of the invention. One embodiment provides a method of modulating the function and/or expression of LHX2 polynucleotide in a patient's cells or tissues in vivo or in vitro, comprising subjecting such cells or tissues to a length of 5 to 30 nucleotides The oligonucleotide is contacted, wherein the nucleotide has at least 5% sequence identity with the reverse complement of the polynucleotide, and the polynucleotide comprises nucleotides 1 to 2145 of SEQ m NO: , nucleotides 1 to 6〇6 in SEQ ID NO: 3, and 5 to 30 contiguous nucleotides in nucleotides 1 to 480 of SEQ ID NO: 4, thereby modulating the patient's cells in vivo or ex vivo The function and/or performance of the LHX2 polynucleotide in the tissue. In one embodiment, the oligonucleotide targets a natural antisense sequence of the LHX2 polynucleotide (eg, the nucleotides set forth in SEQ ID NOS: 2 to 4), and any variants, alleles thereof , homologs, mutants, derivatives, fragments and complementary sequences. Examples of antisense oligonucleotides are, for example, SEq ID NO: 5 to 16 7f> ° Further embodiments provide for the function and/or expression of LHX2 polynucleotides in a patient's cells or tissues in vivo or in vitro. A method comprising contacting the cells or tissues with an antisense oligonucleotide of 5 to 30 nucleotides in length, wherein the nucleotide and the reverse complement of the LHX2 polynucleotide antisense transcript Having at least 50% sequence identity; thereby modulating the function of LHX2 polynucleotides in a patient's cells or tissues in vivo or in vitro and/or Table 156342.doc 201143782. Another embodiment provides a method of modulating the function and/or expression of an LHX2 polynucleotide in a cell or tissue of a patient in vivo or in vitro, comprising subjecting the cell or tissue to a length of 5 to 30 nucleotides An antisense oligonucleotide contact, wherein the oligonucleotide has at least 50% sequence identity to an antisense oligonucleotide of an LHX2 antisense polynucleotide; thereby modulating a patient cell or in vivo or in vitro The function and/or performance of the LHX2 polynucleotide in the tissue. In one embodiment, the composition comprises one or more antisense oligonucleotides that bind to a sense and/or antisense LHX2 polynucleotide. In one embodiment the 'oligonucleotide comprises one or more modified or substituted nucleotides. In one embodiment, the oligonucleotide comprises one or more modified linkages. In a consistent embodiment, the modified nucleotide comprises a modified test group comprising the following: a thioformatate, a phosphonic acid methyl vinegar, a peptide nucleic acid, a 2, a fluorenyl group, a fluorine _ or a carbon, a methylene group or the like. Locked nucleic acid (LNA) molecules. Preferably, the modified nuclear acid-locking nucleic acid molecule comprises a_L-LNA. In one embodiment, the oligonucleotide is administered to the patient subcutaneously, intramuscularly, intravenously or intraperitoneally. In only the examples, nucleotides are administered in the form of a pharmaceutical composition. The treatment regimen includes administering the antisense compound to the patient at least once; however, the treatment can be modified to include multiple doses over a period of time. The treatment can be combined with one or more other types of therapies. In an embodiment, the oligonucleoside is encapsulated in a liposome or attached to a carrier molecule (e.g., cholesterol, TAT peptide). 156342.doc 201143782 Other aspects are set out below. [Embodiment] Several aspects of the present invention will be described hereinafter with reference to the application of the example of the 庑田水. Understand that the texts listed in this article have a complete understanding of the present invention. The present invention may be practiced without the use of any specific details or other methods. The invention is not limited to the order of the various acts or events, which may be The occurrence of different sequences and/or simultaneous occurrences with other actions or events does not necessarily require all of the illustrated actions or events to implement the methods. All of the genes, gene names, and gene products disclosed herein are intended to correspond to the compositions disclosed herein. And methods of homologs from any species H. These terms include, but are not limited to, human and mouse genes and gene products. It should be understood that when revealing genes or gene products from a particular species, this indication is intended to have only The examples are not to be construed as limiting, unless the context clearly indicates. Thus, for example, in certain embodiments relating to mammalian nucleic acid and amino acid sequences, the genes disclosed herein are intended to encompass other animals. Homologous and/or orthologous genes and gene products 'These other animals include, but are not limited to, other mammals, Fish, amphibians, reptiles, and birds. In one embodiment, the gene or nucleic acid sequence is human. Definitions The terms used herein are used for the purpose of describing particular embodiments only and are not intended to limit the invention. The singular forms "a", "a", "a", "a", "the", "the", "the" In addition, the terms "including", "inciudes", "having", "has", "with", or "the" are used in the detailed description and/or the scope of the application. In the case of variations, the terms are intended to mean a range of inclusions in a manner similar to the term "comprising". The term "about" or "approximately" means an acceptable margin of error for a particular value determined by those skilled in the art 'which depends in part on the measurement or measurement of the value', ie, the limitations of the measurement system. . For example, according to industry practice '"about" can mean within one or more standard deviations. Another option of "about" may mean a difference of up to 2%, preferably up to 10°/. More preferably up to 5%, and even more preferably up to 1%. Another option is that, particularly for a biological system or process, the term can mean within an order of magnitude, preferably within 5 times, and more preferably within 2 times of a numerical value. In the case of specific values stated in the scope of the application and the patent application, unless otherwise stated, the term "about" is intended to mean within the acceptable tolerance of the particular value. The term "mRNA" as used herein refers to a currently known mRNA transcript of a dry gene, and any other transcript that can be elucidated. "Antisense nucleus acid" or "antisense compound" means an RNA or DNA molecule that binds to another RNA or DNA (target RNA, DNA). For example, if an RNA oligonucleotide is used, it binds to another RNA target via rnA-RNA interaction and alters the activity of the target RNA. Antisense oligonucleotides can up-regulate or down-regulate the expression and/or function of a particular polynucleotide. This definition is intended to encompass 156342.doc 201143782 containing any foreign RNA or DNA molecule that can be used therapeutically, diagnostically, or otherwise. Such molecules include, for example, antisense RNA or DNA molecules, interfering RNA (RNAi), microRNAs, decoy RNA molecules, siRNA, enzyme RNA, therapeutically editable RNA and RNA agonists and antagonists, antisense oligomeric compounds, Antisense oligonucleotides, external leader sequence (EGS) oligonucleotides, alternative splicing, primers, probes, and other oligomeric compounds that hybridize to at least a portion of the target nucleic acid. Thus, the compounds can be introduced as single-stranded, double-stranded, partially single-stranded, or cyclic oligomeric compounds. In the context of the present invention, the term "oligonucleotide" refers to an oligo or polymer of ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) or a mimetic thereof. The term "oligonucleotide" also encompasses natural and/or modified monomers or linked linear or cyclic oligomers, including deoxyribonucleosides, ribonucleosides, substituted and alpha-rotated Format, peptide nucleic acid (PNA), locked nucleic acid (LNA), phosphorothioate, decyl phosphonate, and the like. Oligonucleotides can specifically bind to a target polynucleic acid by means of a regular pattern of monomer-to-monomer interactions, such as Watson-Crick type base pairing, Hodgsteen type base pairing or reverse Ho6gsteen type base pairing, or And so on. The raised nucleotides can be "chimeric" oligonucleotides, i.e., composed of different regions. In the context of the present invention, a "chimeric" compound is an oligonucleotide comprising two or more chemical regions, such as a DNA region, an RNA region, a PNA region, and the like. In the case of an oligonucleotide compound, each chemical region consists of at least one monomer unit (i.e., nucleotide). Such oligonucleotides typically include at least one region that modifies the oligonucleotide to reveal one or more desired properties. Desirable properties of the oligonucleotide include, but are not limited to, 156342.doc 201143782 (for example): increasing resistance to nuclease degradation, increasing cellular uptake, and/or adding binding affinity for dry nucleic acids. Different regions of the oligonucleotide may thus have different properties. The chimeric nucleotides of the present invention can be formed into a mixed structure of two or more of the above-mentioned nucleotides, modified oligonucleotides, oligonucleosides, and/or nucleotide analogs. • The nucleus acid can be composed of a region that can be joined in a “register”, that is, a single system continuous connection (such as in natural DNA), or a gap ◎ 胄. The spacers are intended to form a covalent "bridge" between the regions and have a length of no more than about one carbon atom in the preferred shape. The spacer may have different functions, such as having a positive or negative charge, having a specific nucleic acid purity (chimeric agent, groove binder, toxin, fluorophore, etc.), having pro-monthly) Grade structure (eg, induction of alanine-containing peptides in a helix). As used herein, "LHX2" and "coffee homologous sequence 2" encompass all family members, mutants, alleles, fragments, species, coding and non-coding Q sequences, sense and antisense polynucleotide chains, and the like. As used herein, the terms UM homologous sequence 2, LHX2, hLhx2, homologous sequence 2 LH2, LH-2, LIM/homologous sequence Lhx2, LIM/homologous sequence 2, MGC138390 are considered identical in the literature, and In case - interchangeable use. As used herein, the term "H. ~.." has specific nucleotides" or "targeted oligonucleotides" and refers to oligonucleotides having the following sequences: (i) capable of Forming a stable complex with a portion of the stem gene, or (1)) capable of forming a stable duplex with a portion of the RNA transcript. The stability of the complex and 156342.doc 201143782 duplex can be calculated by theory and / Or in vitro assays are performed. An example analysis for determining the stability of hybridization complexes and duplexes is set forth in the Examples below. The term "target nucleic acid" as used herein encompasses DNA, RNA (including mRNA precursors) transcribed from the DNA. And mRNA), and cDNA derived from the RNA, coding sequence, non-coding sequence, sense or antisense polynucleotide. The specific hybridization of an oligomeric compound to its target nucleic acid interferes with the normal function of the nucleic acid. This modulation of the function of the target nucleic acid by a compound that specifically hybridizes to the target nucleic acid is commonly referred to as "antisense." The DNA function to be interfered with includes, for example, copying and transcription. The RNA function to be interfered with includes all important functions, such as shifting of RNA to protein translation sites, translation of proteins from RNA, splicing of RNA to produce one or more mRNA species, and catalytic activity in which RNA can participate or promote. The overall effect of this interference on the function of the target nucleic acid is the regulation of the expression of the encoded product or oligonucleotide. RNA interference ("RNAi") is mediated by double-stranded RNA (dsRNA) molecules with sequence-specific homology to the "target" nucleic acid sequence. In certain embodiments of the invention, the medium is a "small interference" RNA duplex (siRNA) of 5-25 nucleotides. The siRNA is derived from the treatment of dsRNA by an RNase enzyme called Dicer. The siRNA duplex product is recruited into a polyprotein siRNA complex (RNA-induced silencing complex) called RISC. Without wishing to be bound by any particular theory, it is believed that RISC can be directed to a target nucleic acid (mRNA is preferred) where the siRNA duplex interacts in a sequence-specific manner to catalyze cleavage in a catalytic manner. Small interfering RNAs which can be used in the present invention can be synthesized and used according to procedures well known in the art and known to those skilled in the art, 156342. doc -10- 201143782. The small interfering RNA used in the methods of the invention suitably comprises from about 1 to about 50 nucleotides (nt). In an example of a non-limiting embodiment, the siRNA can comprise from about 5 to about 4, about 5 to about 3,000, about 10 to about 30 nt, about 15 to about 25 拊, or about 2 〇. _25 nucleotides. The selection of appropriate oligonucleotides is facilitated by the use of computer programs that automatically align nucleic acid sequences and indicate regions of identity or homology. These programs are used to compare the obtained nucleic acid sequences by, for example, searching a database such as GenBank or by measuring the pCR product. Comparison of nucleic acid sequences from various species allows selection of nucleic acid sequences that show the degree of agreement between appropriate species. In the case of unsequenced genes, Southern blotting is performed to determine the degree of agreement between the target species and other species. Consistency can be roughly measured by performing Southern blots (as is well known in the industry) with varying degrees of stringency. Such procedures allow for the selection of oligonucleotides that exhibit a high degree of complementarity to the target nucleic acid sequence in the individual to be controlled and a lower degree of complementarity to the corresponding nucleic acid sequences in other species. Those skilled in the art will recognize that a wide range of gene regions suitable for use in the present invention can be selected. "Enzyme RNA" means an RNA molecule with enzymatic activity (Cech, (1988) J.

American. Med· Assoc. 26〇, 3030-3035). The enzymatic nucleic acid (ribose, enzyme) first acts by binding to the target RNA. The binding is carried out via a dry binding portion of an enzymatic nucleic acid that is immediately adjacent to the enzymatic portion of the molecule used to lyse the dry RNA. Thus, the enzymatic nucleic acid first recognizes the target RNA and then binds to the target rNA via the genomic pairing and acts enzymatically to cleave the target RN A upon binding to the exact site. 156342.doc 201143782 "Decoy RNA" means an RNA molecule that mimics the natural binding domain of a ligand. The decoy RNA thus competes with the natural binding target for binding to a specific ligand. For example, it has been shown that overexpression of HIV transactivation (TAR) RNA can be used as a "bait" and effectively binds to the HIV tat protein, thereby preventing its binding to the TAR sequence encoded in HIV RNA. This is intended to be a specific example. Those skilled in the art will recognize that this is only an example and that other embodiments can be readily derived using techniques generally known in the art. The term "monomer" as used herein generally means attached by a phosphodiester bond or an analog thereof to form an oligomer having a size ranging from a plurality of monomer units (for example, about 3-4) to about several hundred monomer units. A monomer of a nucleotide. Phosphodiester linked analogs include: phosphorothioates, dithiophosphates, methyl phosphonates, phosphatidyl phosphates, amine phosphates, and the like, as described more fully below. The term "nucleotide" encompasses naturally occurring nucleotides as well as non-naturally occurring nucleotides. Those skilled in the art should be aware that the various nucleotides previously considered "non-natural" have subsequently been discovered in nature. Therefore, "nucleotide" includes not only molecules containing a known quaternary ring and a heterocyclic hexacyclic ring, but also heterocyclic analogs and tautomers thereof. An illustrative example of other types of nucleotides are molecules that contain: adenine, guanine, thymine, cytosine, uracil, guanidine, xanthine, diamine guanidine, 8-sided oxy-Ν6-曱Adenine, 7-deazapurine, 7-deazaguanine, N4, N4-ethanol cytosine, N6, N6-ethanol-2,6-diaminopurine, 5-quinone cytosolic, 5-(C3-C6)-alkynyl group. dense. 5-, 5-fluorouridine α-bite, 5-bromoindoline, pseudoisopyrimidine, 2-hydroxy-5-methyl-4-triazolopyridine, isocytosine 156342.doc -12- 201143782 "Non-naturally occurring" nucleotides as described in U.S. Patent No. 5,432,272 to Benner et al. The term "nucleotide" is intended to cover every and all such examples as well as the analogs and tautomers thereof. Nucleotides of particular interest include adenine, guanine, thymine, cytosine, and uracil, which are considered naturally occurring nucleotides associated with therapeutic and diagnostic applications in humans. Nucleotides comprise native 2'-deoxy and 2-hydroxyl sugars (for example, as described in Kornberg and Baker, DNA Replication, 2nd Edition (Freeman, San Francisco, 1992)) and analogs thereof are related to nucleotides. An "analog" comprises a synthetic nucleotide having a modified base moiety and/or a modified sugar moiety (see, for example, as outlined below: Scheit, Nucleotide Analogs, John Wiley, New York, 1980;

Freier & Altmann, (1997) Nucl. Acid. Res., 25(22), 4429-4443, Toulme, JJ, (2001) Nature Biotechnology 19:17-18; Manoharan M·, (1999) Biochemica et Biophysica Acta 1489:117-139 i Freier SM, (1997) Nucleic Acid Research, 25:4429-4443, Uhlman, E., (2000) Drug Discovery & Development, 3: 203-213, Herdewin P., (2000) Antisense & Nucleic Acid Drug Dev" 10: 297-310); 2'-0, 3'-C linked [3.2.0] bicyclic glucosinolates. These analogs are designed to enhance binding properties (eg, Synthetic nucleotides of duplex or triplex stability, specificity, or the like. As used herein, "hybridization" means the pairing of substantially complementary strands of an oligomeric compound. A pairing mechanism involves hydrogen bonding between complementary nucleosides or nucleotides (nuclear acid) of a compound chain, which can be Watson-Crick, 156342.doc • 13-201143782 HWgsteen or reverse H〇dgSteen Hydrogen bond. For example, adenine and thymidine are complementary nucleotides that form a hydrogen bond pair. Hybridization can occur without circumstances. 5 An antisense compound is "specifically hybridizable" when the binding of the compound to the target nucleic acid interferes with the normal function of the target nucleic acid to modulate the function and/or activity' and is sufficiently complementary to avoid antisense compounds The non-native nucleic acid sequence undergoes non-specific binding under conditions where it is desired to undergo heterosexual binding (also under physiological conditions in the case of in vivo or therapeutic treatment, and under conditions in which the analysis is performed in the case of in vitro analysis). As used herein, the phrase "stringent hybridization conditions" and "stringent conditions" refer to the conditions under which a compound of the invention hybridizes to its target sequence, but with a minimum number of: sequence hybridizations. Strict conditions are sequence dependent and in different cases and In the context of the present invention, the "stringent conditions" of the heteropoly compound and the target sequence are determined by the nature and composition of the polycondensation compound and the analysis thereof. In general, stringent hybridization conditions include low concentrations (<0.15 M) of low ionic strengths of salts containing inorganic cations (e.g., Na++ or κ++), temperatures higher than that of milk, and less than oligomeric compounds: target sequence complexes Tm of the substance, and the presence of a denaturant (such as formamidine, dimethylformamide monomethyl sulfoxide) or detergent sodium dodecyl sulfate (SDS). For example, 5' decreases the rate of hybridization for every -1% metformin. An example of high stringency hybridization conditions is O.i χ sodium chloride-sodium citrate buffer (ssc) / o.i% (w/v) SDS (3 sec at 6 crc). As used herein, "complementary" refers to the ability to precisely pair between two nucleotides on one or two oligomeric chains. For example, if the antisense compound 156342.doc -J4- 201143782 is placed in a nucleobase and can be verified at a position near the target nucleic acid (the nucleic acid DNA, RNA, or oligodeoxynucleotide molecule) When hydrogen bonding occurs, the position at which hydrogen bonding occurs between the oligonucleotide and the target nucleic acid is regarded as a complementary position. When a sufficient number of complementary positions in each molecule are occupied by a nucleoside that can be hydrogen bonded to each other, the 'oligomer compound and other DNA, RNA, or oligonucleotide' molecules are complementary to each other. Thus, "specifically hybridizable" and "complementary" are used to indicate a term for a sufficient number of nucleotides to have a sufficient degree of exact pairing or complementarity such that I is stable between the oligomeric compound and the target nucleic acid and Specific binding. It is understood in the art that the oligomeric compound sequence need not be 100% complementary to its target nucleic acid sequence to be specifically hybridized. Alternatively, the raised nucleotides can hybridize in one or more regions such that the inserted or adjacent segments do not participate in hybridization events (e.g., loop structures, mismatches, or hairpin structures). The polycondensation compound of the present invention comprises at least about 7%, or at least about 75%, or at least about 8%, or at least about 85%, or at least about 9%, or about the target region of the target nucleic acid sequence to which it is dried, or To ◎ about 95% less, or at least about 99% sequence complementarity. For example, an antisense compound of 18 of the 20 nucleotides of the antisense compound that is complementary to the target region and thereby specifically hybridizes will represent a 99% complementarity. In this example, the remaining non-complementary nucleotides may be clustered or dispersed with complementary nucleotides and need not be adjacent to each other or adjacent to the complementary nucleotides. Thus, an antisense compound of 18 nucleotides in length and having 4 (four) non-complementary nucleotides flanked by two regions that are fully complementary to the target nucleic acid has 77.8% overall complementarity to the target nucleic acid. It is within the scope of the invention. The complementation of antisense compounds with a target nucleic acid region can generally be measured using the BLAST program known in the art (base partial pair 156342.doc -15-201143782 quasi-search tool) and PowerBLAST program. The homology, sequence identity, or percent complementarity can be used by, for example, the Gap program (Wisc〇ns Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, Madison Wis.) using default settings (its use) Smith and Waterman algorithm) (Adv Appl. Math., (1981) 2, 482-489) As used herein, the term "thermodynamic melting point (Tm)" means drying under defined ionic strength, pH, and nucleic acid concentration. The temperature of the sequence complementary to the oligonucleotide that hybridizes to the target sequence under equilibrium. Generally, for short-collar acid (for example, having 1 〇 to 5 核苷酸 nucleotides), stringent conditions are the following: salt concentration is at least about (1) (seven) to ^ MiNa ion concentration at pH 7.0 to 8.3. (or other salt) and at a temperature of at least about 3 (TC ^ stringent conditions can also be achieved by the addition of a destabilizing agent such as guanamine. As used herein, "modulating" means increasing (stimulating) or reducing (inhibiting) a gene. In the context of a polynucleotide sequence, the term "variant" may encompass a polynucleotide sequence associated with a wild-type gene. This definition may also include, for example, "alleles", Splice, "species" or "polymorphism" variants. Splice variants can be significantly consistent with a reference molecule, but usually have a larger or smaller number of aggregates due to alternate splicing of exons during mRNA processing. Nucleotide. The corresponding polypeptide may have an additional functional domain or a non-existing domain. The sequence of the variant of the species varies from species to species. It is particularly useful in this month to be a variant of the wild-type gene product. Body At least one mutation in the nucleic acid sequence, and can produce altered mRNA or a polypeptide whose structure or function may or may not be altered. Any 'given natural or = cause: does not have an allele Gene forms, common mutational changes with 1 or many allelic variants are usually attributed to the day of the nucleus: loss, addition, or substitution. Each of these changes can be alone in the sequence of the foot. Or in combination with others—or multiple times. The resulting polypeptides typically have significant amino acid-likeness with each other. Polymorphisms change in the nucleotide sequence of a particular gene between individuals in a given species.

Polymorphic variants may also cover "single nuclear bud acid polymorphism" (sNp) or single-killer canine changes, in which the base of the polymorphic acid sequence changes. The presence of a SNP may indicate, for example, that a population has a predisposition to disease state (i.e., susceptibility to resistance). Beta Polynucleotide comprises undergoing chemical modification (eg, hydrogen is replaced by an alkyl, alumina or amine group) Nucleic acid. Derivatives (e.g., derived oligonucleotides) can be used. Non-naturally occurring parts, such as altered sugar moieties or intersaccharide linkages. Examples of μ, such as derivative phosphorothioates, and other sulfur-containing material-derived nucleic acids known in the art may also contain a label, including radionucleotides, enzymes, phosphor chemiluminescent agents, color formers, receptors, Cofactors, inhibitors, magnetic particles, and the like. Derivatives of peptides or peptides, for example, are modified by: glycosylation, polyethylation, phosphating, sulfating, reduction/alkylation, thiolation, chemical coupling, or mild flu Marlin (formalin) processing. The derivative may also be modified to contain a detectable label (directly or indirectly) including, but not limited to, a radioisotope, a fluorescent, and an enzymatic label. The term "animal" or "patient" as used herein is intended to include, for example, human 156342.doc. •17· 201143782 Mammals, wolves, rats, birds, chickens, reptiles, sheep, elk, deer, mule deer, water, horse, cow, pig, goat, dog, cat, rat, animal, fish, Insects and J. Spiders. "Mammals" include warm-blooded mammals (eg, humans and domestic animals) that are usually under medical care. Examples include cats, horses, cows, and humans, and only humans. Or treatment" encompasses treatment of a disease state in a mammal, and comprises: (4) preventing the disease state in a mammal, particularly when the mammal is susceptible to a disease state but has not been diagnosed with the disease state (b) inhibiting the disease state, for example, preventing its development; and/or (c) slowing the disease state, for example, reducing the disease state until the desired endpoint is reached. It also includes improving the symptoms of the disease (for example, alleviating pain or discomfort), where the improvement may directly affect or may not directly affect the disease (eg, cause, infection, performance, etc.). "Neural disease or condition" as used herein means Any of the diseases or conditions of the nervous system and/or the visual system. "Nervous diseases or conditions" include 2 central nervous systems (brain, brainstem and cerebellum), peripheral nervous system (including cranial nerves), and autonomic nervous system (located A disease or condition of the central and peripheral nervous system. The neurological disease or condition includes, but is not limited to, acquired epilepsy aphasia; acute disseminated encephalomyelitis; adrenal leukodystrophy, age-related macular degeneration Symptoms; dysplasia; aphasia; Aicardi syndr〇me; Alexander disease; Alpers' disease; crossed limb dysentery Alzheimer's disease (Alzheimer's disease) Vascular dementia; 156342.doc -18- 201143782 Amyotrophic lateral sclerosis; no brain malformation; angel syndrome (Angeiman) Syndrome), hemangiopathy; dystrophy; aphasia; apraxia; arachnoid cyst, arachnoiditis; An 〇 _ _ ( ( ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; 'Syndr〇me); ataxia telangiectasia; attention deficit with • motility disorder; autism; autonomic dysfunction; back pain; Batten disease, Beckett's disease Behcet, sdisease); BeU'S Palsy; benign essential sputum; benign local muscle atrophy; benign intracranial pressure, Binswanger disease (Binswanger, s disease); eyelid climbing; Brock-Suz B1〇ch Sulzberger syndrome; brachial plexus injury; brain abscess; brain injury; brain tumor (including glioblastoma multiforme); spinal cord tumor; Brown Sequard syndrome; Canavan disease; wrist pain syndrome; burning neuralgia; central pain syndrome; central medullary cerebral lysis, cerebral aneurysm; cerebral arteriosclerosis; cerebral atrophy; brain giant Cerebral palsy; Charcot-Marie-Tooth disease; chemotherapy-induced neuropathy and neuropathic pain; Chiad malformati〇n; chorea; chronic inflammatory degeneration Myelinating polyneuropathy; chronic pain; chronic regional pain syndrome group 'ke _ er 2 syndrome (to produce ^ L〇 Wry syndrome); coma 'contains persistent vegetative state; congenital facial paralysis; cortical basal degeneration' Gu arteritis, craniosynostosis; Creutzfeldt-Jakob dlSeaSe; accumulated traumatic disease; Cushing's syndrome; giant cell inclusion disease; cytomegalovirus infection; dance eye - 156342. Doc •19- 201143782 Dance foot syndrome; Dandy Walker syndrome; Dawson disease; De Morsier's syndrome; cloned Peck-Clump's syndrome (Dejerine- Klumke palsy); dementia; dermatomyositis; diabetic neuropathy; diffuse sclerosis; autonomic dysfunction; difficulty in writing; dyslexia; dystonia; early childhood epileptic brain ; empty sella syndrome; encephalitis; cerebral palsy; brain trigeminal ganglion tumor; epilepsy; Erb's palsy; idiopathic seismic; Fabry's disease; (Fahr's syndrome); fainting; familial spastic paralysis; febrile seizures; Fisher syndrome; Friedreich's ataxia; frontotemporal dementia and other r tau lesions Gaucher's disease; Gerstmann's syndrome; giant cell arteritis; giant cell inclusion disease; globular white matter dystrophy; Guillain-Barre Syndrome); HTLV-1 associated myelopathy; Hallervorden-Spatz disease; head injury; headache; hemifacial spasm; hereditary spastic paraplegia; hereditary polyneuritis-like ataxia; Herpes zoster; herpes zoster; Hirayama syndrome; HIV-associated dementia and neuropathy (and eight 11) neurological manifestations; forebrain non-cracking malformation; Huntington's disease (Huntington, s (10) )and Other polyglutamine indole repeat disease; water-free no brain malformation; hydrocephalus; hypercortisolism; hypoxia; immune-mediated encephalomyelitis; inclusion body myositis; 2 dysregulated disease; Disease; infant reefsum disease (four) coffee to refsum disease) · 'baby sputum; inflammatory myopathy; intracranial cyst; intracranial 156342.doc -20- 201143782 high pressure; Jubbert syndrome (Joubert syndrome); Keams-Sayre syndrome; Kennedy disease; Kinsboume syndrome; Klippel Feil syndrome; Krabbe disease Kugelberg-Welander disease; Kuru disease; Lafora disease; Lambert-Eaton myasthenic syndrome; Landa - Landau-Kleffner syndrome; lateral medullary (Wallenberg) syndrome; learning disability; Leigh's disease; Lennox-Gastau syndrome (Lennox-Gustaut Syndrome); Lai-naphthalene Lesch-Nyhan syndrome; leukodystrophy; Lewy body dementia; no cerebral gyrus; atresia syndrome; Lou Gehrig's disease (ie, motor neuron disease or Amyotrophic lateral sclerosis), lumbar disc disease; Lyme disease - neurological sequelae, Machado-Joseph disease; brain hypertrophy, giant brain; Mel-Rosenthal syndrome Menieres disease; Meningitis; Menkes disease; Metachromatic leukodystrophy; Microcephaly; Migraine; Miller Fisher syndrome ; small stroke; mitochondrial myopathy; Mobius syndrome; single limb muscle atrophy; motor neuron disease; abnormal vascular network disease of the brain; mucopolysaccharidosis; multiple infarct dementia; Focal motor neuropathy; multiple sclerosis and other demyelinating disorders; multiple systemic atrophy with positional hypotension; muscular dystrophy; myasthenia gravis; demyelinating diffuse sclerosis; infant muscle 156342.doc -21- 201143782 Clonic encephalopathy; myoclonus; myopathy; myotonia; narcolepsy; neurofibromatosis; neuroleptic malignant syndrome; neurological manifestations of AIDS; neurological sequelae of lupus; neuromuscular rigidity; neuronal waxy lipid Leptin; abnormal brain neuron migration; Niemann_pick disease; O'Sullivan-McLeod syndrome; occipital neuralgia; recessive spinal canal insufficiency sequence Ohtahara syndrome; cerebral atrophy of cerebral palsy; strabismic cerebral palsy; optic neuritis; orthostatic hypotension; overuse syndrome; paresthesia' neurodegenerative disease or disorder (Parkinsen's disease) 〇n, s disease), utton's disease, Alzheimer's disease, amyotrophic lateral sclerosis (ALS), dementia, multiple sclerosis and other diseases and conditions associated with neuronal cell death); congenital deputy Myotonia; paraneoplastic disease; paroxysmal episode, Pary R〇mberg (Pelizaeus-Merzbacher disease); periodic 瘫痪 'periphery Trans disease; painful neuropathy and neuropathic pain; persistent vegetative state; systemic developmental delay; optical sneezing reflex; phytanic acid storage disease; Pick's disease; nerve kneading; pituitary tumor; polymyositis ; brain penetrating malformation; polio pruritus syndrome; postherpetic neuralgia; post-infectious encephalomyelitis; orthostatic hypotension; Prader-Willi syndrome; primary lateral sclerosis ; prion disease; progressive one-sided atrophy; progressive multifocal leukoencephalopathy; progressive sclerosing gray matter atrophy; progressive nucleus itching; pseudocephaloma; Ramsay-Hunt syndrome (Types I and II); Rasmussen's encephalitis; Reflex sympathetic dystrophy 156342.doc •22· 201143782 Syndrome; Refsum disease; Repetitive motor disorder; Reproducibility Compression injury; restless leg syndrome; retrovirus-associated myelopathy; Rett syndrome; Rey's syndrome; chorea (Saint Vitus dance) ); Sandhoff disease; Schilder's disease; cerebral palsy; clear septum optic nerve dysplasia; scare infant syndrome; herpes zoster; Shay-Dreig syndrome (Shy -Drager syndrome); Sjogren's syndrome; sleep apnea; Soto's syndrome; spasticity; spina bifida; spinal cord injury; spinal cord tumor; spinal muscular atrophy; stiff syndrome (Stiff-Person Syndrome; stroke; Sturge-Weber syndrome; subacute sclerosing panencephalitis; subcortical arteriosclerotic encephalopathy; Sydenham chorea; syncope; syringomyelia Delayed dyskinesia; Tay-Sachs disease; Temporal arteritis; Spinal cord syndrome; Thomsen disease; Thoracic outlet syndrome; Tic Douloureux; Todd Todd's paralysis; hyperactive slang syndrome; transient ischemic attack; disseminated spongiform encephalopathy; transverse myelitis; traumatic brain injury; Trigeminal neuralgia; tropical spastic paraplegia; tuberous sclerosis; vascular dementia (multiple infarct dementia); vasculitis, including temporal arteritis; Von Hippel-Lindau disease; Wallenberg's syndrome; Werdnig-Hoffman disease; West syndrome; cervical spine; Williams syndrome; Wilson 156342.doc -23- 201143782 Disease (Wildon's disease); and Zweiger syndrome syndrome) ° "A fluid disease or condition" contains diseases, conditions, or conditions that affect hematopoietic cells or tissues. Blood disorders include diseases, conditions, or conditions associated with abnormal blood levels or function. Examples of blood disorders include conditions derived from bone marrow irradiation or cancer chemotherapy, such as the following conditions: pernicious anemia, hemorrhagic anemia, hemolytic anemia, aplastic anemia, sickle cell anemia, iron granule anemia, Anemia associated with chronic infections (eg malaria, trypanosomiasis, HI V, hepatitis virus or other viruses), myeloid anemia caused by bone marrow loss, renal failure from anemia, anemia, polycythemia, infectious singles Nuclear cell hyperplasia (IM), acute non-lymphocytic leukemia (ANLL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), acute myelomonocytic leukemia (AMMoL), true erythrocytosis Disease, lymphoma, acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia, wilms, s turn〇r, Ewing's sarcoma, retinoblastoma, hemophilia, and Symptoms associated with increased blood test risk, cytotherapy, thalassemia, and antibody-induced conditions (eg, transfusion reactions and erythroblastia) ) mechanical trauma of red blood cells (eg, microangiopathic hemolytic anemia, thrombotic thrombocytopenic purpura, and disseminated intravascular coagulation), infections caused by parasites (eg, protozoa), from, for example, lead poisoning Chemical damage, and spleen function. Lymphatic diseases include, but are not limited to, lymphadenitis, lymphatic vessel dilatation, lymphangitis, lymphedema, lymphocystosis, lymphoproliferative disorders, cutaneous mucosal lymph node syndrome, reticuloendothelial proliferation, spleen 156342.doc -24- 201143782 Disease, thymic hyperplasia, thymic tumor, tuberculosis, lymph nodes, pseudo lymphoma, and lymphoid abnormalities. Conditions of the lymphoid hematopoietic system include, but are not limited to, non-Hodgkin's lymphoma, chronic lymphocytic leukemia, and reactive lymphoid tissue hyperplasia. • Polynucleotide and Nucleotide Compositions and Molecules • Targets: In one embodiment, the target comprises a nucleic acid sequence of LIM homolog 2 (LHX2), including (without limitation) senses associated with LHX2 and/ Or antisense non-coding and / or coding sequences. The LIM-homology domain (LIM-HD) protein regulates the tissue specificity of the target gene and is required for the normal development and maintenance of many organs. During embryonic development of vertebrates and invertebrates, members of the LIM homology domain transcription factor family play an important role by controlling processes such as asymmetric cell division, tissue specificity, and differentiation of specific cell types. The member of this family, Lhx2, is particularly interesting based on its function in the development of several different tissues. Its function is achieved through interstitial-epithelial interactions and regulation of stem/Q progenitor cells. A member of this family of Lhx2 lineage transcription factors, which consists of 12 known genes, which are structurally characterized by two amino-terminal zinc finger motifs (LIM domains) and a carboxy-terminal homology domain. Inactivation of Lhx2 in mice results in a decrease in cerebral cortex size and abnormal development of the striate, telencephalon, olfactory system, and dermal pattern formation.

Lhx2 is also required for normal limbs and liver formation; Lhx2-/- mice develop liver fibrosis and incomplete erythropoiesis, leading to death from severe anemia before birth. The known locus of Lhx2 expression is extremely close to these phenotypes 156342.doc •25- 201143782. Lhx2 is expressed in the liver-associated primitive transverse mesenchyme that is part of the liver and maintains its performance during liver development until the mature stage of hepatic stellate cells. The Lhx2-/- embryo showed a decrease in the size of the liver that had been shown at E1〇5, indicating that Lhx2 is required for fetal liver expansion. Mutant expression is derived from the presence of activated hepatic stellate cells to produce fibrotic and schizophrenic cells containing phenotypically abnormal endoderm cells. The interstitial defects in the liver of Lhx2_/_ mice cause fatal anemia with cell involuntaryness, as Lhx2-/- hematopoietic cells appear normal, suggesting that the mutant microenvironment does not support hematopoietic development. These findings indicate that Lhx2 in hepatic stellate cells interacts with mesenchymal-epithelial cells, and these interactions are important for liver expansion, organization, differentiation, and formation of hematopoietic microenvironments in fetal liver. In one embodiment, an antisense oligonucleotide is used to prevent or treat a disease or condition associated with a tribute to the LHX2 family. An example um homologous sequence 2 (LHX2) mediated disease and disease mediated by stem cell regeneration obtained from an antisense compound can be used (4): associated with an abnormal uterine b and/or performance of the rib Disease or condition, blood disease or condition, impaired hair growth, neurological disease or condition, retinal disease or condition, disease or condition associated with immunization, pituitary disease or condition, liver disease or condition, smell " Money or illness, impaired erythropoiesis, creation & disease or condition, meningococcal disease and vertebral limb length. In % of cases, one or more antisense oligonucleotides are used to regulate LHX2 in patients in need , thereby preventing or treating any disease or condition associated with the LHX2 abnormality table, activity (compared to the normal control group). 156342.doc -26 - 201143782 In one embodiment, the nucleotides are gathered for LHX2 Nucleotides are specific 'which include, without limitation, non-coding regions. LHX2 targets include variants of LHX2; mutants of LHX2, including SNp; LHX2i non-coding sequences; alleles, fragments and the like. Preferably, the oligonucleotide is an antisense RNA • molecule. According to an embodiment of the invention, the target nucleic acid molecule is not limited to the LHX2 polynucleotide but extends to any isoform, receptor, homologue, non-LHX2 A coding region and the like. In one embodiment, the oligonucleotide targets a natural antisense sequence (a natural sequence encoding a non-coding region) of the LHX2 target, including, without limitation, variants, alleles thereof, Homologs, mutants, derivatives, fragments and complementary sequences. Preferably, the nucleotides are antisense RNA or DNA molecules. In a consistent application, the oligomeric compound of the invention is also included in the compound. Or a variant of a different base at a plurality of nucleotide positions. For example, if the first nucleotide is adenine, it may be produced at this position containing a chest, a bird, a cytidine or other natural or non- Variants of natural nucleotides. This can occur anywhere in the antisense compound. These compounds are then tested using the methods described herein to determine their ability to inhibit the performance of a target nucleic acid. In some embodiments, antisense compounds and The same between targets The sex, sequence or complementarity is from about 5% to about 6%. In some embodiments, the homology, sequence identity or complementarity is from about 6% to about 7%. In some embodiments The 'homology, sequence identity or complementarity is from about 7% to about 80%. In some embodiments, homology, sequence identity or complementarity is from about 80°/ to about 90% » in In some embodiments, homology, sequence identity 156342.doc -27- 201143782 or complementarity is about 90%, about 92%, about 94%, about 95°/, about 96%, about 97%, about 98%, about 99% or about 100%. The antisense compound can specifically hybridize in the following cases: the binding of the compound to the dry nucleic acid can interfere with the normal function of the target nucleic acid resulting in loss of activity, and there is sufficient complementarity to avoid Non-specific binding of the antisense compound to the non-target nucleic acid sequence under conditions where it is desired to specifically bind. Such conditions include (i.e., physiological conditions in the case of in vivo analysis or therapeutic treatment, and conditions under which the analysis is performed in the context of in vitro analysis. An antisense compound (whether DNA, RNA, chimeric compound, or substituted compound, etc.) can specifically hybridize in the following situations: binding of the compound to a target DNA or RNA molecule can interfere with the normal function of the target DNA or RNA. Loss of utility, and sufficient degree of complementarity to avoid antisense compounds and non-target sequences under conditions that are expected to specifically bind (ie, under physiological conditions in the case of in vivo analysis or therapeutic treatment, or in vivo) Non-specific binding occurs under conditions in which the analysis is performed in the case of an external analysis. In one embodiment, targeting LHX2 (including, without limitation, antisense sequences identified and expanded using, for example, PCR, hybridization, etc., one or more of the sequences set forth in SEQ ID NOs: 2 to 4, and the like ) can adjust the performance or function of LHX2. In one embodiment, the performance or function is up-regulated compared to the control group. In one embodiment, the performance or function is downregulated as compared to a control group. In one embodiment, the oligonucleotide comprises a nucleic acid sequence as set forth in SEQ ID NOS: 5 to 16, comprising an antisense sequence identified and expanded using, for example, PCR, hybridization, and the like. Such oligonucleotides can include one or more modified nucleotides, shorter or longer fragments, modified linkages, and the like. Modified bond or nucleoside 156342.doc -28 - 201143782 dithiophosphoric acid or such a derivative. Examples of linkages that can be attached to the present acid include thiophthalic acid vinegar, dithiophenes. In an embodiment, the nucleotide comprises a phosphorus derivative. The phosphorus derivative (or modified phosphate group) of the sugar or sugar analog moiety of the bright red nucleoside I acid may be a monophosphate, a diphosphate, a sulfonium triphosphate, an alkane Phosphates, phosphorothioates, and the like. The preparation of the analogs of the above-mentioned analogs, as well as the methods of their introduction into nucleotides, modified nucleotides and oligonucleotides, are also known per se and need not be described herein. ❹ v et al. For this technology, the specificity and sensitivity of antisense oligonucleotides are also utilized in therapeutic applications. Antisense oligonucleotides are used as therapeutic moieties to treat disease states in animals and humans. Antisense oligonucleotides have been safely and efficiently administered to humans and many clinical trials are currently being performed. It is thus determined that the raised nucleotides can be a useful therapeutic modality that can be used to treat cells, tissues, and animals, particularly in human therapeutic regimens. In embodiments of the invention, oligomeric antisense compounds, particularly nucleus Binding to a target nucleic acid molecule and modulating the expression and/or function of the molecule encoded by the target gene. The DNA function to be interfered with includes, for example, replication and transcription, and the RNA function to be interfered with includes all important functions, such as RNA to protein. Translocation of translation sites, translation of proteins from RNA, splicing of one or more mRNA species by Rna, and catalytic activity that RNA can participate in or promote. Up-regulation or inhibition of function can be performed depending on the desired function. , or a cyclic polymeric compound antisense compound comprising an antisense oligomeric compound, an antisense scorpion acid 'external leader sequence (EGS) oligonucleotide, an alternative splicing, a primer, a probe, and others A polymeric compound that hybridizes to at least a portion of a nucleic acid.

156342.doc -29- 201143782 Introduction. The targeting of an antisense compound to a particular nucleic acid molecule can be a multi-step process in the context of the present invention. This process often begins with the identification of the dry nucleic acid of the regulated function. For example, the dry nucleic acid can be a nucleic acid molecule that exhibits a cellular gene (or mRNA transcribed from the gene) associated with a particular disorder or disease state or from an infectious agent. In the present invention, (iv) acid-encoded um homologous sequence 2 (LHX2). The dry-direction process also often includes the determination of at least one target region, region, or site that produces an antisense interaction within the nucleic acid to produce a desired effect (e.g., modulate expression). Within the context of the present invention, the term "region" is defined as a portion of a target nucleic acid having at least one distinguishable structure, function, or property. There is a segment within the region of the target nucleic acid. A "segment" is defined as a small or sub-portion of a region within a target nucleic acid. The "site" used in the present invention is defined as the position within the dry nucleic acid. In one embodiment, the antisense oligonucleotide binds to the natural antisense sequence of UM homolog 2 (LHX2) and regulates the expression and/or function of LHX2 (SEq m N〇: u. Examples of antisense sequences include ID N〇: 2 to 16. In one embodiment, the 'antisense oligonucleotide binds to one or more segments of the LIM homolog 2 (LHX2) polynucleotide and modulates LHX2i expression and/or功月 b.区# further comprises at least 5 contiguous nucleotides of LHX2 sense or antisense polynucleic acid. In one embodiment, the antisense oligonucleotide is specific for the natural antisense sequence of LHX2, The binding of the raised nucleotide to the natural antisense sequence of LHX2 can modulate the expression and/or function of LHX2. 156342.doc • 30· 201143782 In one embodiment, the oligonucleotide compound comprises SEQ ID NO: 5 to The sequence of 16 is identified and expanded using, for example, pcR, hybridization, etc. The oligonucleotides may include one or more modified nucleotides, shorter or longer fragments, modified keys. And the like. Examples of modified bonds or internucleotide linkages include phosphorothioates, dithiocarbamate or the like. In one embodiment, the nucleotide comprises a phosphorus derivative. It can be attached to the hair

The phosphorus derivative (or modified phosphate group) of the sugar or sugar analog moiety in the modified modified nucleotide may be a monophosphate, a diphosphate, a trican acid ester, an alkyl phosphate, an alkanoic acid. Esters, phosphorothioates, and the like. The preparation of the above-described phosphate vinegar analogs, and their incorporation in nucleotides, modified nucleotides and oligonucleotides are also known per se and need not be set forth herein. As is known in the art, 'the translation initiation codon is usually 5I_AUG (in the transcriptional mRNA molecule; in the corresponding DNA molecule, 5, _ATG), the translation initiation codon is also called "AUG codon", "start codon" Or "AUG starter horse", number gene. Has a translation start codon, which has rn a sequence 5, -GUG, 5i_UUG5l5i_CUG; and has been shown ^_AUA, 5, _acg and 5, -CUG can be played in vivo effect. Therefore, even if the starting amino acid is in each case, it is usually methionine (in eukaryotes) or methionine (in prokaryotes), the term "translation start codon" and "starting" Codons can also cover a variety of codon sequences. Eukaryotic and prokaryotic genes may have two or more alternative initiation codons, one of which may be preferentially used for translational initiation in a particular cell type or group, in a tissue, or under a particular set of conditions. . In the context of the present invention, the "starting codon" and "turning-start codon" are used to transcribe the gene in the (4)-moving self-encoding L paste source sequence 156342.doc -31- 201143782 column 2 (LHX2). One or more codons of the translation of the mRNA 'regardless of the sequence of the codons. The gene translation stop codon (or "stop codon") can have three sequences (ie, 5, Uaa, 5'-UAG, and 5'-UGA, and the corresponding DNA sequences are 5, -TAA, 5, -TAg, respectively). And one of the 5'-TGA). The terms "start codon region" and "translation initiation codon region" refer to any mRNA or gene in the direction from the translation initiation codon (ie, 5 or 3) covering from about 25 to about 5 A portion of a neighboring nucleotide. Similarly, the terms "stop codon region" and "translatation stop codon region" refer to the orientation or orientation (ie, 5' or 3') of the mRNA or gene from the translation stop codon that covers about 25 to about A portion of 5 adjacent nucleotides. Therefore, the "start codon region" (or "translation start codon region") and the "stop codon region" (or "transition stop codon region") are all regions in which the antisense compound of the present invention can effectively be used. An open reading frame (〇RF) or "encoding region" known in the art refers to the region between the translation start codon and the translation stop codon, which is also an area that can be effectively targeted. Within the context of the present invention, the targeting region encompasses the intragenic region of the translational start or stop codon of the open reading frame (ORF) of the gene. Another target region comprises the 5, non-translated region (5, utr) known in the art, which refers to the portion of the mRNA from the direction 5 of the translation initiation codon, and thus comprises the 5' capping site of the mRNA. And translating the nucleotides (or corresponding nucleotides in the gene) between the initiation codons. Another target region comprises the 3, non-translated region (3'UTR) known in the art, which refers to the mRNA portion from the translation stop codon 3, 156 342. doc -32 - 201143782 up and thus contains mRNA Translate the stop codon and the nucleotide between the 3' ends (or the corresponding nucleotide on the gene). The 5' capping site of the mRNA includes a 5'-most residue N7-mercaptoguanosine residue joined to the mRNA via a 5'-5' triphosphate linkage. The 5' capped region of the mRNA is considered to comprise the 5' capping structure itself and 50 nucleotides immediately adjacent to the capping site. Another target area of the present invention is a 5. capping area. Although some eukaryotic mRNA transcripts are directly translatable, many eukaryotic mRNA transcripts contain one or more regions that are transcribed and translated as "introns" prior to translation. The remaining (and thus translated) regions are referred to as "exons" and are spliced together to form a continuous mRNA sequence. In one embodiment, a targeted splice site (ie, an 'intron-exon node or an exon-intron node) is particularly useful for aberrant splicing associated with disease, or for excessive splicing products Produce a situation related to the disease. Another embodiment of an abnormal fusion node that is due to rearrangement or loss is a dry site. An mRNA transcript produced by splicing two (or more) mRNAs from different gene sources is referred to as a "fusion transcript." An antisense compound that is light, for example, to a DNA or mRNA precursor, can be used to efficiently bind to an intron. In one embodiment, the antisense raised nucleotide binds to the coding and/or non-coding region of the dry polynucleotide and modulates the performance and/or function of the dry molecule. In one embodiment, the antisense oligonucleotide binds to a natural antisense polynucleotide and modulates the expression and/or function of the target molecule. In one embodiment, the antisense oligonucleotide binds to a sense polynucleotide and modulates the expression and/or function of the target molecule. Alternative RNA transcripts can be produced from the same genomic region of DNA. Alternative transcripts such as 156342.doc •33· 201143782 are commonly referred to as “variants”. More specifically, "mRNA precursor variants" are transcripts produced from the same genomic DNA that differ from the other transcripts produced from the same genomic DNA in terms of starting or stopping positions and contain introns and Proton sequence. When one or more exons or intron regions, or portions thereof, are cleaved during splicing, the mRNA precursor variants produce smaller "mRNA variants". Thus, mRNA variants of treated mRNA precursor variants and each unique mRNA precursor variant must always produce a unique mRNA variant by splicing. Such mRNA variants are also referred to as "alternate splice variants". If the mRNA precursor variant is not spliced, the mRNA precursor variant is identical to the mRNA variant. Variants can be generated via the use of alternating signals that initiate or stop transcription. The mRNA precursor and mRNA may have more than one start codon or stop codon. Variants derived from mRNA precursors or mRNAs using alternative initiation codons are referred to as "alternative initiation variants" of mRNA precursors or mRNA. Their transcripts using alternative stop codons are referred to as "alternative stop variants" of mRNA precursors or mRNA. A specific type of alternative stop variant is a "P〇lyA variant" in which multiple transcripts are derived from an alternative to the transcription machine for one of the "P〇lyA stop signals", resulting in A transcript that terminates at the unique P〇lyA site. In the context of the present invention, the types of variants described herein are also examples of target nucleic acids. The position at which the target nucleic acid hybridizes to the antisense compound is defined at least as a portion of the target region which is 5 nucleotides long by the active antisense compound. Although specific sequences of certain exemplary target segments are set forth herein, those skilled in the art will recognize that such sequences are used to illustrate and elucidate specific implementations within the scope of the disclosure 156342.doc • 34 201143782. example. Those skilled in the art can readily identify other segments 7 according to the present disclosure that are considered to be 5-1 00 nucleotides in length and include a segment of at least five (5) consecutive segments selected from preferred interpretive target segments. The target segment of the nucleotide is also suitable for targeting 0. The target segment may comprise a DNA or RNA sequence comprising at least 5 contiguous nucleotides from the 5 terminus of one of the preferred interpretable target segments (remaining nucleoside The acid is a contiguous stretch of the same DNA or RNA that begins immediately upstream of the 5'-end of the light segment and continues until the sputum or RNA contains from about 5 to about 1 nucleotide. Likewise, preferred target segments are represented as DNA or RNA sequences comprising at least 5 contiguous nucleotides from the 3, _ terminus of one of the preferred interpretable target segments (remaining nucleotides are the same DNA) Or a continuous segment of the RNA that begins immediately downstream of the 3, _ terminal of the target segment and continues until the DNA or RNA contains from about 5 to about 100 nucleotides. The skilled artisan can identify other preferred target segments without undue experimentation. After identifying one or more target regions, segments or sites, the target can be selected to be sufficiently complementary to the target (ie, the hybridization is sufficiently sufficient and sufficiently specific). Antisense compounds to achieve the desired effect. In an embodiment of the invention, the oligonucleotide binds to an antisense strand of a particular target. The nucleotides are at least 5 nucleotides in length and can be synthesized to make each oligo Nucleic acid (4) overlapping sequences, so that the oligonuclear pure synthesis to cover the entire length of the polynuclear acid 4 also contains the m纟 flat code region. In one implementation, the antisense oligo (4) is more specific to the specific nucleic acid 156342.doc -35- 201143782 Sense compound to specific nucleic acid target A multi-step process that often begins with the identification of a nucleic acid sequence that regulates function. This can, for example, be a cellular gene (or mRNA transcribed from a gene) associated with a particular disorder or disease state, or a non-coding polynucleoside. Acids (eg, non-coding RNA (ncRNA)) RNA can be classified into (1) messenger RNA (mRNA), which translates into protein; and (2) non-protein-encoding RNA (ncRNA). ncRNA includes microRNA, antisense Transcripts and other transcription units (TUs) that contain high-density stop codons and lack any extensive "open reading frame". Many ncRNAs appear to start in the 3' non-translated region of the locus encoding the protein (3 The ntRNA is usually rare and at least half of the ncRNAs sequenced by the FANTOM Alliance do not appear to be polyadenylated. Most researchers focus on processing and export to the cytoplasm for obvious reasons. Polyadenylation of mRNA. Recently, the group showing non-polyadenylated nuclear RNA may be extremely large, and many of these transcripts are derived from so-called intergenic regions. ncRNA regulates the mechanism of gene expression and transcripts Base pairing. RNAs that function by base pairing can be grouped into: (1) cis-encoded RNA, which encodes at the same genetic position in which it functions, but encodes at the opposite strand of RNA and thus displays its target Complete complementarity; and (2) trans-encoded RNA, which encodes at a different chromosomal location than the RNA it functions with and generally does not exhibit complete base-pairing potential with its target. Unexpectedly limited by theory, this article Interference with an antisense oligonucleotide against an antisense polynucleotide can alter the expression of the corresponding sense messenger RNA. However, this regulation can be uncoordinated (an increase in messenger RNA caused by knockdown) or coordination (antisense weakening) Causes a decrease in accompanying messenger RNA). In such cases, the anti-156342.doc •36-201143782 sense oligonucleotide can target overlapping or non-overlapping portions of the antisense transcript to create a weakening or sequestration. Both coding and non-coding antisense transcripts can be targeted in the same manner, and any species can modulate the corresponding sense transcript in a coordinated or uncoordinated manner. The strategy employed in identifying new oligonucleotides for use with a target can be based on any other way of attenuating the antisense RNA transcript by antisense oligonucleotides or modulating the desired stem. Nongluo i: In the case of uncoordinated regulation, weakening antisense transcripts will increase the performance of the learned (sense) gene. If the latter gene encodes a known or putative drug target, it is conceivable to attenuate its antisense equivalent to mimic the effects of the receptor agonist or enzyme stimulator. In the case of coordinated regulation, the agricultural strategy can simultaneously weaken the antisense and sense transcripts and thereby reduce the conventional (sense) gene expression. For example, if the anti-sense nucleotide is used to achieve attenuation, then this strategy can be used to apply an antisense oligonucleotide targeting a sense transcript and another antisense targeting the corresponding antisense transcript. Oligonucleotides, or a single energy symmetric antisense oligonucleotide that simultaneously targets overlapping sense and antisense transcripts. According to the invention, an antisense compound comprises an antisense oligonucleotide, a ribozyme, an external leader sequence (EGS) oligonucleotide, an siRNA compound, a single- or double-stranded RNA interference (RNAi) compound (eg, an siRNA compound), and Other polymeric compounds that hybridize to at least a portion of the dry nucleic acid and modulate its function. Thus, it can be DNA, RNA, DNA-like, RNA-like, or a mixture thereof, or can be a mimetic of one or more of such substances. The compounds may be single-stranded double-stranded, cyclic or hairpin poly-compounding compounds and may contain structural elements such as internal or terminal, mismatch or loop. Antisense compounds are typically made 156342.doc • 37· 201143782 Linear n but can be joined or otherwise made into a cyclic and/or branched form. Antisense compounds can comprise, for example, the following constructs: hybridized to form complete Or a double strand of a partially double-stranded compound, or a single strand having sufficient auto-complementary I' to hybridize and & into a fully or partially double-stranded compound. The two chains can be joined internally to create a free 3, or 5, end or connectable to form a continuous hairpin. The hairpin type structure may have a pendant P knife at the M3' end to extend the single chain feature. The double-stranded compound may optionally comprise a pendant knife at the end. The modification may comprise a coupling group attached to one end, a selected nucleotide acid site, a sugar site or attached to a internuclear bud linkage. Alternatively, the two strands can be joined via a non-nucleic acid moiety or a linker group. Upon formation from only one strand, the dsRNA may be in the form of an auto-complementary hairpin-type molecule that folds in itself to form a duplex. Thus, the dsRNA can be either full or sickle double-stranded to specifically modulate @gene expression by stably expressing the dsRNA hairpin 5L in the transgenic cell line, however, in some embodiments, the gene expression or function is upregulated. When a single strand is formed from two strands, or a self-complementary hairpin-type molecule is opened (self-folded to form a duplex), the two strands (or regions in which the duplex forms a duplex) are Wats〇n_Crick mode base pairing complementary RNA strand. Following introduction into the system, the compounds of the invention may elicit the action of one or more enzymes or structural proteins to effect cleavage or other modification of the target nucleic acid or may act via a mechanism based on occupancy. In general, nucleic acids (including oligonucleic acids) can be described as "DNA-like" (ie, usually having one or more 2,_deoxy sugars and (through*)T instead of u bases) or "classes" rnA" (ie, usually having one or more 2'-hydroxy or 2,-modified sugars and (usually} υ instead of τ bases). Nucleic acid snail 156342.doc -38- 201143782 can be used in more than one structure Types, most commonly type A and type B. It is believed that, in general, nucleotides having a B-like structure are "like DNA" and those having a type A structure are "like RNA". (Chimeric) In embodiments, the antisense compound may contain A- and B-type regions. In one embodiment, the desired oligonucleotide or antisense compound comprises at least one of the following: antisense RNA, antisense DNA , chimeric antisense oligonucleotides, including modified linkages of antisense oligonucleotides, interfering RNA (RNAi), short interfering RNA (siRNA); micro interfering RNA (miRNA); small temporal RNA (stRNA); Or short hairpin RNA (shRNA); small RNA-induced gene activation (RNAa); small activating RNA (saRNA), or a combination thereof. dsRNA can also activate gene expression, a mechanism called Small RNA-induced gene activation" or RNAa. The dsRNA targeting the gene promoter induces efficient transcriptional activation of related genes. RNAa is displayed in human cells using synthetic dsRNA (called "small activating RNA" (saRNA). It is not known whether RNAa is conserved in other organisms. It has been found that small double-stranded RNA (dsRNA) (such as small interfering RNA (siRNA) and microRNA (miRNA)) is triggered by the evolutionary conservation mechanism of RNA interference (RNAi). RNAi typically causes gene silencing by remodeling chromatin to thereby repress transcription, thereby degrading complementary mRNA, or blocking protein translation. However, in the context of the examples section below, oligonucleotides may be displayed. Increase the expression and/or function of the LIM homolog 2 (LHX2) polynucleotide and its encoded product. The dsRNA can also be used as a small activating RNA (saRNA). It is not expected to be limited by theory, by the gene promoter In the sequence, saRNA can induce target gene expression, a phenomenon known as dsRNA-induced transcriptional activation (RNAa) of 156342.doc-39-201143782. In another embodiment, the "better dry segment" identified herein is available Screening for additional compounds that modulate the expression of LIM homolog 2 (LHX2) polynucleotides. "Modulators" are compounds that reduce or increase the performance of a nucleic acid molecule encoding LHX2' and include at least a complementary 5-nucleotide portion of the segment. The screening method comprises the steps of: contacting a preferred segment of a nucleic acid molecule encoding a lhx2 sense or natural antisense polynucleotide with one or more candidate modulators, and selecting One or more candidate modulators that reduce or increase the performance of a nucleic acid molecule encoding an LHX2 polynucleotide (eg, SEq m N〇: 5 to 16). If one or more candidate modulators are shown to be capable of modulating (eg, reducing or increasing) the performance of a nucleic acid molecule encoding an LHX2 polynucleotide, the modulator can be used in other investigative studies of LHX2 polynucleotide function' or as a A research, diagnostic, or therapeutic agent of the invention. Targeting a natural antisense sequence preferably modulates the function of the target gene. For example, the LHX2 gene (for example, accession number is NM_0 (M789). In one embodiment, the antisense polynucleotide of the LHX2 gene is killed. In one embodiment, the antisense oligonucleotide targets the LHX2 polynucleus. Sense and/or natural antisense sequences, variants, alleles, isoforms, homologs, canine variants, street organisms, fragments and complementary sequences of a nucleoside (for example, accession number _004789). Preferably, the nucleotide is raised as an antisense molecule and the target comprises an antisense and/or a sensed [coding and non-coding region of the Ηχ2 polynucleotide. The preferred segment of the invention may also be complementary to the present invention. Antisense compounds are combined to form stable double-stranded (duplex) oligonucleotides. It has been shown in the industry that the double-stranded oligonucleotides such as 3H can regulate the expression of 耙 and pass 156342.doc •40- 201143782 by The sense mechanism regulates translation and RNA processing. Additionally, the double-stranded portion can undergo chemical modification. For example, it has been shown that the double-stranded portion can inhibit the target by typical hybridization of the antisense strand of the duplex with the target, thereby Initiating enzymatic degradation of the target. In one embodiment, the antisense oligonucleotide targets the LIM homolog 2 (LHX2) polynucleotides (e.g., accession number NM_004789)), variants, alleles, isoforms, homologs, mutants, derivatives, fragments, and complementary sequences. Preferably, the oligonucleotide is an antisense molecule. According to an embodiment of the invention, the target nucleic acid molecule is not limited to LHX2 but extends to any of the isoforms, receptors, homologs, and the like of the LHX2 molecule. In one embodiment, the oligonucleotide targets a natural antisense sequence of the LHX2 polynucleotide (eg, the polynucleotides set forth in SEQ ID NOS: 2 to 4), any variant thereof, etc. Genes, homologs, mutants, derivatives, fragments and complementary sequences. Examples of antisense oligonucleotides are set forth in SEQ ID NOS: 5 to 16. In one embodiment, the oligonucleotide complements or binds to the nucleic acid sequence of the LHX2 antisense molecule (including, without limitation, non-coding sense and/or antisense sequences associated with the LHX2 polynucleotide) and modulates the LHX2 molecule Performance and / or function. In one embodiment, the oligonucleotide complements or binds to the nucleic acid sequence of the LHX2 natural antisense molecule (as set forth in SEQ ID NOS: 2 to 4) and modulates the expression and/or function of the LHX2 molecule. In one embodiment, the raised nucleotide comprises the sequence of at least 5 contiguous nucleotides of SEQ ID NOs: 5 to 16 and modulates the expression and/or work of the LHX2 molecule. 156342.doc -41 - 201143782 Polynucleotide Target These include LHX2 (including its family members), variants of LHX2; mutants of LHX2, including SNPs; non-coding sequences of LHX2; alleles of LHX2; species variants, fragments and the like. Preferably, the oligonucleotide is an antisense molecule. In one embodiment, the oligonucleotides that target the LHX2 polynucleotide include: antisense RNA, interfering RNA (RNAi), short interfering RNA (siRNA); micro interfering RNA (miRNA); small temporal rna (stRNA) Or short hairpin RNA (shRNA); small RNA-induced gene activation (RNAa); or small activating RNA (saRNA). In one embodiment, 'targeting LIM homologous sequence 2 (LHX2) polynucleotides (e. g., SEQ ID NOs: 2 to 16) can modulate the expression or function of such targets. In one embodiment, the performance or function is up-regulated compared to the control group. In one embodiment, the performance or function is downregulated as compared to a control group. In one embodiment, the antisense compound comprises the sequences set forth in SEQ ID NOs: 5 to 16. Such nucleotides may include one or more modified nucleotides, shorter or longer fragments, modified linkages, and the like. In one embodiment, SEQ ID NOS: 5 to 16 comprise one or more LNA nucleotides. Table 1 shows exemplary antisense oligonucleotides for use in the methods of the invention. Table 1 ·· Sequence number Antisense sequence name sequence SEQ ID NO: 5 CUR-1560 (^*T*(^*(^*(^*T*G*G*G*As|iC!*T*G* T*A*A*G*G*A SEQ ID NO: 6 CUR-1561 156342.doc •42· 201143782

SEQ ID NO: 7 CUR-1562 T*了*γ*(^*γ*(^*Τ*0*(^*€Ι*0*0*(^*Τ*ΟΙ*Τ*0*0*0 ^*Τ SEQ ID NO: 8 CUR-1563 SEQ ID NO: 9 CUR-1588 q*T*T*G*T*A*T*G*T申G*G*G*T*TT*T*G *G*G*T*G*G SEQ ID NO: 10 CUR-1589 As,eC*A*T*C*C*C*T*G3|£C*T*Ti,:T*C*T* C*Cs|iC*T*G SEQ ID NO: 11 CUR-1590 T*T*T*A*G*T*A*G*T*A*C*C*A*C*C*G*G *A*G*C*C SEQ ID NO: 12 CUR-1591 C*C*C*A*G*C*T*T*C*A*T*C*C*T*C*A*G* A*G*C SEQ ID NO: 13 CUR-1592 T*T*G*G*G*T*T'*Gs|err*G*A*G*G*G*G*A*G G *A SEQ ID NO: 14 CUR-1593 T*C*C*A*T*A*T*C*C*A*G*C*C*G*T*G*C*C*A*C* T SEQ ID NO: 15 CUR-1594 T*c*C*A*T*T*T*G*T*C*A*C*C*A*C*C*A*G*C*A*C SEQ ID NO: 16 CUR-1595 G*G*C*T*T*C*T*G*G*G*T*G*TsliT*T*C*G1,iT*T5|eT

The desired target nucleic acid can be modulated in a number of ways known in the art. For example, antisense oligonucleotides, siRNA, and the like are used. Enzymatic nucleic acid molecules (e.g., ribozymes) are nucleic acid molecules capable of catalyzing one or more different reactions, including the ability to repeatedly cleave other individual nucleic acid molecules in a nucleotide base sequence specific manner. The enzymatic nucleic acid molecule can be used, for example, to target substantially any RNA transcript. Due to their sequence specificity, trans-cleaved enzymatic nucleic acid molecules can be used as a therapeutic agent for human diseases. Enzymatic nucleic acid molecules can be designed to cleave specific RNA targets within the cellular RNA background. This cleavage event causes the mRNA to lose functionality and remove protein expression from the RNA. In this way, the synthesis of proteins associated with disease states can be selectively inhibited. In general, an enzymatic nucleic acid having an RNA cleavage activity functions by first binding to a target RN A . This binding is carried out via a target binding portion of an enzymatic nucleic acid which is in close proximity to the enzymatic portion of the molecule used to cleave #RNA. Thus, the enzymatic nucleic acid first recognizes the target RNA and then binds to the target RNA via a complementary base pair and acts enzymatically to cleave the target RNA upon binding to the exact site. This cleavage strategy for a target RNA will disrupt its ability to direct synthesis of the encoded protein by 156342.doc-43-201143782. After the enzymatic nucleic acid has bound and cleaves its RNA target, it is released from the RNA to find another target and can repeatedly bind and cleave the new target. Several methods, such as in vitro selection (evolution) strategies (〇rgel, (1979) pr〇c. R. Soc. London, B 205, 435), have been used to generate various reactions (eg, phosphodiester linkages and hydrazines). A new nucleic acid catalyst for the cleavage and linkage of amine linkages. The development of ribozymes with optimal catalytic activity will significantly contribute to any strategy that uses RNA lytic ribozymes to regulate gene expression. For example, hammerhead ribozymes act at a catalytic rate of about 1 min-丨 in the presence of a saturated (10 mM) concentration of Mg2+ cofactor. Artificial "RNA ligase" ribozymes have been shown to catalyze the corresponding self-modification reactions at a rate of about 1 〇〇. In addition, certain modified hammerhead ribozymes are known to have a binding arm composed of DNA that catalyzes RNA cleavage by a multiple of turnover rate. Finally, the preparation of certain nuclear (iv) analogs in place of the specific residues in the nucleus catalyzed core of the head-like ribozyme produces carboxylase, which shows a catalytic rate that is increased by up to a factor of 1. These findings indicate that ribozymes can significantly promote chemical conversion by significantly greater than the catalytic rate of the rate of destruction exhibited by most native self-cleaving ribozymes in vitro. The structure of certain self-cleaving ribozymes can be optimized for maximum catalytic activity, or a novel RNA motif that exhibits a significantly faster rate of RNA phosphodiester cleavage can be prepared. The intermolecular cleavage of the rna-bearing catalyzed by the RNA catalyst catalyzed by the "hammerhead" model was first shown in 1987 (10) enbeck, 〇c (1987) 328. 596_6〇〇) mNA catalyst and made it with the rural rna molecule 156342 .doc -44 - 201143782 Reaction, thus indicating that it is indeed catalytic. Catalytic RNA based on the "hammerhead" motif design has been used to cleave specific target sequences by making appropriate base changes in the catalytic RNA to maintain the necessary base pairing with the target sequence. This allows the use of catalytic RNA to cleave specific target sequences and demonstrates that catalytic RNA designed according to the "hammerhead" model can cleave specific receptor RNA in vivo. RNA interference (RNAi) has become an effective tool for regulating gene expression in mammalian and mammalian cells. This approach requires the delivery of small interfering RNA (siRNA) in the form of RNA itself or in the form of DNA using expression plasmids or viruses and coding sequences for small hairpin RNA for processing into siRNA. This system enables efficient delivery of siRNA precursors to their active cytoplasm and allows the use of regulated and tissue-specific promoters for gene expression. In one embodiment, the oligonucleotide or antisense compound comprises a polymer or polymer of ribonucleic acid (RNA) and/or deoxyribonucleic acid (DNA), or a mimetic, conjugate, analog or homolog thereof Things. The term encompasses oligonucleotides consisting of naturally occurring nucleotides, sugars, and covalent internucleoside (backbone) linkages; and oligonucleotides having non-naturally occurring portions and functioning in a similar manner. Such modified or substituted oligonucleotides are often preferred over natural forms due to desirable properties such as enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases. According to the invention, an oligonucleotide or "antisense compound" comprises an antisense oligonucleotide (eg RNA, DNA, mimetic, chimera, analog or homolog thereof), ribozyme, external leader sequence (EGS) Oligonucleotides, siRNA compounds, single- or double-stranded RNA interference (RNAi) compounds (eg, siRNA compound 156342.doc-45-201143782), saRNA, aRNA, and other six-nucleus with target nucleic acid) A main v is an oligomeric compound that is partially heterozygous and has its function. Therefore, its RNA, DNA, rNa, or i mixture *' 〇ΝΑ 'A, a mouthpiece, or may be a mimic of - or more of these. The compounds may be single bond, double chain, cyclic or hairpin type oligomeric compounds and may contain structural elements such as internal or terminal expansion, mismatch or loop. Antisense compounds are typically made in a linear form, but may be joined or otherwise formed into a cyclic and/or branched form. The antisense compound may comprise, for example, the following (four): two strands that are hybridized to form a fully or partially double-stranded compound, or a single strand that has sufficient auto-complementarity to hybridize and form a fully or partially double-stranded compound. The two strands can be joined internally to create a free 3' or 5' end or can be joined to form a continuous hairpin type structure or loop. The hairpin type structure may have an overhang at the end of 5, or 3 to extend the single-strand character. The double-stranded compound may contain an overhang at the end as needed. Other modifications may include a coupling group attached to one end, a selected nucleotide position, a sugar position, or a linkage to a nuclear tone. Alternatively, the two strands can be joined via a non-nucleic acid moiety or a linker group. When formed from only one strand, the dsRNA can be in the form of an auto-complementary hairpin-type molecule that folds in itself to form a duplex. Thus the 'dsRNA can be fully or partially double stranded. Gene expression can be specifically regulated by stably expressing the dsRNA hairpin type in the transgenic cell line. When formed from a single strand of two strands, or in the form of an autorecognition hairpin-type molecule (self-folding to form a duplex), the two strands (or regions in which the duplex forms a duplex) are Watson- Crick-type base pairing complementary RNA strand. Following introduction into the system, the compounds of the invention may elicit one or more enzymes or cleaves to effect cleavage or other modification of the t-nucleic acid or may act via a mechanism based on occupancy. In general, nucleic acids (including nucleus) can be described as "DNA-like" (ie, usually with one or more deoxy sugars and (usually) τ instead of u-test) or "RNA-like" (ie, , usually with one or more 2, a trans- or 2'-modified sugar and (usually) U instead of a T-test). Nucleic acid spiro = privately available - more than one type of structural class, most commonly type A and type B. As a general example, in the case of a nucleotide-like "DNa" having a B-like structure and having an A-like structure, the "antigen-like compound" may contain a_& Type b area. Antisense compounds of the invention can include an antisense portion of from about 5 to about 8 nucleotides in length (i.e., ' from about 5 to about 8 such linked nuclear buds). This refers to the length of the antisense stock or part of the antisense. In other words, the single-bond antisense compound of the present invention comprises 5 to about 8G cores (four), and the double-stranded antisense compound (phantom of dsRNA) of the present invention comprises a sense and a counter of 5 to about 8 nucleotides in length. A stock or part. Those skilled in the art should understand that this cover length is 5, Ο 6 19 31 43 55 67 79 20 32 44 56 68 , 9 21 33 45 57 69 10 22 34 46 58 70 11 23 35 47 59 71 12 24 36 48 60 72 13 25 37 49 61 73 14 26 38 50 62 74 15 27 39 51 63 75 16 28 40 52 64 76 17 29 41 53 65 77 18 30 42 54 66 78 , or 80 nucleotides, or any of them The antisense part of the scope. In an embodiment, the antisense compound of the invention has an antisense portion of from 1 to 5 nucleotides in length. Those skilled in the art should understand that this shows the reverse 156342.d〇 (-47· 201143782 meaning part length is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 , 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47 An oligonucleotide of 48, 49, or 50 nucleotides, or any range thereof. In some embodiments, the oligonucleotide is 15 nucleotides in length. In one embodiment, the invention The antisense or oligonucleotide compound has an antisense portion of 12 or 13 to 30 nucleotides in length. Those skilled in the art will appreciate that the length of the antisense portion is 12, 13, 14, 15, 16 , 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 nuclear acid, or any range of antisense compounds thereof. In one embodiment, The oligomeric compounds of the invention also comprise variants in which different bases are present at one or more nucleotide positions in the compound. For example, if the first nucleotide is adenine, it may be produced. The position contains a variant of thymidine, guanosine or cytidine. This can occur anywhere in the antisense or dsRNA compound. These compounds are then tested using the methods described herein to determine their ability to inhibit the performance of the target nucleic acid. In embodiments, the homology, sequence identity or complementarity between the antisense compound and the target is from about 40% to about 60%. In some embodiments, the homology, sequence identity or complementarity is about 6 〇% to about 7%. In some embodiments, homology, sequence identity or complementarity is from about 7% to about 80%. In some embodiments, homology, sequence identity or complementarity From about 80% to about 90%. In some embodiments, homology, sequence identity or complementarity is about 90%, about 92%, about 94%, about 95%, about 96%, about 156,342.doc -48· 201143782 97%, about 98°/, about 99% or about 100%. In one embodiment, antisense oligonucleotides (for example, the nucleic acid molecules described in SEQ ID NOS: 5 to 16) Included in one or more substitutions or modifications. In one embodiment, the nucleotide is substituted with a locked nucleic acid (LNA). • In one embodiment, the oligonucleotide One or more regions of a sense and/or antisense nucleic acid molecule that encodes a coding and/or non-coding sequence associated with LHX2 and sequences as set forth in SEQ ID NOs: 1 to 4. Oligonucleotides also target SEQ ID NO: overlap region of 1 to 4. 〇U Certain preferred oligonucleotides of the invention are chimeric oligonucleotides. In the context of the present invention, "oligonucleotide" or "complex" is an oligonucleotide comprising two or more chemically distinct regions, each region consisting of at least one nucleotide. The oligonucleotides typically contain at least one modified nucleotide acid region that confers one or more beneficial properties (eg, increased nuclease resistance, increased cellular uptake, increased binding affinity to the target); and is capable of cleaving RNA : DNA or RNA: the region of the RNA heterozygote that is regulated by the enzyme. 0 For example, RNase Η is an endonuclease that cleaves the RNA strand of an RNA:DNA duplex. Thus, activation of RNase Η cleaves RNA targets, thereby greatly enhancing the efficiency of antisense regulation of gene expression. Thus, comparable results are generally obtained with shorter oligonucleotides when using chimeric-oligonucleotides compared to phosphorothioate deoxyoligonucleotides that hybridize to the same 'target region'. Cleavage of the RNA target can typically be detected by gel electrophoresis and, if desired, related nucleic acid hybridization techniques known in the art. In one embodiment, the chimeric oligonucleotide comprises at least one region modified to increase target binding affinity, and (often) used as a region for RNAse Η receptor. The affinity of an oligonucleotide to its target (in this case, a nucleic acid encoding ras), Tm-based oligonucleotides, is typically determined by measuring the Trn of the oligonucleotide/target pair 156342.doc-49-201143782. The temperature at which the target dissociates; spectrophotometry is used to detect dissociation. The higher the Tm, the greater the affinity of the oligonucleotide to the target. The chimeric antisense compounds of the invention may be formed as a composite structure of two or more oligonucleotides, modified oligonucleotides, oligonucleosides and/or oligonucleotide mimetics as described above. Thus, compounds are also known in the art as hybrids or combinations. Representative U.S. patents that teach the preparation of such hybrid structures include, but are not limited to, U.S. Patent Nos. 5, 〇13, 8:3 、, 丨^, 77, 5, 220, 〇〇7 胄, 5, 256, 775. No. 5,366,878, 5,403,711, 5,491,133, 5,565,35, 5,623,065, 5,652,355, 5,652,356, and 5,700,922, each of which is incorporated herein by reference. In one embodiment, the modified region of the oligonucleotide comprises at least one nucleotide modified at the 2' position of the sugar, preferably via a 2,-0 alkyl, 2,-0-alkyl group. - 〇-alkyl or 2'-fluoro modified nucleotide. In another embodiment, the RNA modification comprises 2' to the ribose, abasic residue or the reverse base of the 3' end of the pyrimidine Fluorine, 2,-amino group and 2, 〇-methyl modification. These modifications are usually incorporated into the oligonucleotide and the oligonucleotides have been shown relative to the 2,-deoxyoligonucleoside I for a given The target has a higher Tm (i.e., higher target binding affinity). This increased affinity effect greatly enhances the inhibition of gene expression by RNAi oligonucleotides. An endonuclease that cleaves an RNA strand in an rNA:dna duplex: thus, activation of this enzyme cleaves the rna stem, and thereby greatly enhances the efficiency of RNAi inhibition. The cleavage of the RNA target is usually performed by 156342. Doc •50· 201143782 Gel electrophoresis for display. In one embodiment, chimeric nucleotides are also modified to enhance nuclease resistance. Cells contain a variety of degradable nucleic acid exonuclease and endonuclease A variety of nucleotide and nucleoside modifications have been shown to allow oligonucleotides incorporating such modifications to be more resistant to nuclease digestion than native oligodeoxynucleotides. Nuclease resistance is typically achieved by oligonucleosides The acid is incubated with the cell extract or the isolated nuclease solution and is often measured by gel electrophoresis over a period of time to measure the amount of remaining intact oligonucleotide. Oligonucleotides that have been modified to enhance their nuclease resistance Glycosides remain intact for longer than unmodified oligonucleotides. A variety of oligonucleotide modifications have been shown to enhance or confer nuclease resistance. Currently, oligonucleotides containing at least one phosphorothioate modification are preferred. In some Nucleotide-enhancing nucleotides that enhance target binding affinity can also independently enhance nuclease resistance. Specific examples of some preferred nucleotides to be used in the present invention include those including modified backbones, The modified backbones are, for example, phosphorothioates, phosphotriesters, decyl phosphonates, short chain alkyl or cycloalkyl hydrazine linkages or short chain heteroatoms or heterocyclic saccharide linkages. Preferred are oligonucleotides having a phosphorothioate backbone and those having a hetero atom backbone, such as CH2--NH--0--CH2, CH, -N ( CH3)--0--CH2 [referred to as fluorenylene (mercaptoimine) or MMI backbone], CH2--0--N(CH3)--CH2, CH2-N(CH3)--N (CH3)--CH2 and 0--N(CH3)--CH2--CH2 backbone, wherein the natural phosphodiester backbone is represented by 0--P--0--CH). The guanamine backbone disclosed by De Mesmaeker et al. (19'95) Acc. Chem. Res. 28:366-374 is also preferred. Also preferred are oligonucleotides having a morpholino backbone structure (Summerton and Weller, U.S. Patent No. 5,034,506) in another example, 156, 342. doc - 51, 201143, 782 (for example, peptide nucleic acid (PNA) master Chain), the phosphodiester backbone of the oligonucleotide is replaced by a polyamine backbone, which binds directly or indirectly to the aza nitrogen atom of the polyamine backbone. The raised nucleotides may also include one or more substituted sugar moieties. Preferred nucleotides include one of the following groups at the 2' position: OH, SH, SCH3, F, OCN, 0CH30CH3, OCH3 0(CH2)n CH3, 0(CH2)n NH2 or 0(CH2) n CH3 (n is from 1 to about 10); C1-C10 lower alkyl, alkoxy, substituted lower alkyl, alkaryl or aralkyl; Cl; Br; CN; CF3; OCF3; , S--, or N-alkyl; 〇__, S--, or N-alkenyl; SOCH3; S02 CH3; 0N02; N02; N3; NH2; heterocycloalkyl; heterocycloalkylaryl; Alkylamino group; polyalkylamino group; substituted decyl group; RNA cleavage group; reporter group; immigration agent; group for improving the pharmacokinetic properties of nucleotide recruitment; A group that improves the pharmacodynamic properties of an oligonucleotide and other substituents having similar properties. A preferred modification comprises 2'-methoxyethoxyethoxy [2LO-CH2 CH2 OCH3, also known as 2-0-(2-decyloxyethyl)]. Other preferred modifications include 2'-decyloxy (2'-0--CH3), 2'-propoxy (2'-OCH2 CH2CH3) and 2'-fluoro (2'-F). Similar modifications can be made at other positions on the nucleotide, particularly at the 1' position of the 3' terminal nucleotide and at the 5' position of the 5' terminal nucleoside. The nucleotides may also have a sugar mimetic, for example, using cyclobutyl instead of the pentofuran. Nucleotides may also be modified or substituted, in addition or alternatively as a nucleobase (commonly abbreviated as "base"). As used herein, "unmodified" or "natural" nucleotides include adenine (A), guanine (G), thymine (T), cytosine (C), and uracil (U). The modified nucleotide comprises only 156342.doc -52 - 201143782 or a nucleotide found in the natural nucleic acid, such as hypoxanthine 6 methyl adenine, 5-Me pyrimidine, especially 5-methyl Pyrimidine (also known as 5-mercapto-deoxycytosine and commonly referred to in the industry as 5_Me-C), 5-based on mouth-bite (HMC), glycosyl-based HMC, and gentio-disaccharide-based HMC; ^External 夂 成 nucleation bud • Acid, for example, 2_amino adenine, 2-(methylamino) adenine, 2_(imidazolylalkyl) adenine, 2_(alkylalkylamino) Adenine or other hetero-substituted alkyl adenine, 2-thiouracil, 2-thiothymidine, 5-bromouracil, Hong ◎ hydroxymethyl uracil, 8-azaguanine, 7-azepine guanine , 6 (6-aminohexyl) adenine and 2,6-diaminopurine. "Common" bases known in the art (e.g., inosine) can be included. The 5_Me-C substitution has been shown to increase the stability of the nucleic acid duplex by 0.6-1.2. (: and is currently a preferred base substitution. Another modification to the nucleotides of the present invention involves chemically linking one or more of the conjugates that enhance the activity or cellular uptake of the raised nucleotide to the recruitment Nucleotides "These moieties include, but are not limited to, lipid moieties (eg, cholesterol moieties), cholesteryl moieties, aliphatic chains (eg, dodecanediol or ◎ undecyl residues), polyamines or poly(ethylene) Alcohol chains, or adamantane acetic acid. Nucleotides comprising lipophilic moieties, and methods of making such oligonucleotides are known in the art, for example, U.S. Patent Nos. 5,138, No. 45, No. 5,218,1 〇5 and 5,459,255. All positions in a given nucleotide need not be uniformly modified, and in fact one or more of the above modifications may be incorporated into a single oligonucleotide or even incorporated into a nucleoside In the single nucleoside of the acid, the present invention also encompasses the nucleus acid which is conjugated to the above-mentioned nucleotides. In another embodiment, the nucleic acid molecule of the present invention is coupled to another moiety, 156342. Doc -53· 201143782 This other part contains but not limited to Abasic nucleotides, polyethers, polyamines, polyamines, peptides, carbohydrates, lipids, or polyhydrocarbon compounds. Those skilled in the art will recognize that such molecules can be attached to sugars, The number of positions on the base or phosphate vine group includes one or more of any of the nucleic acid molecules of the nucleic acid molecule. The oligonucleotides used in the present invention can be prepared in a convenient conventional manner via well-known solid phase synthesis techniques. Equipment for carrying out the synthesis is sold by a number of suppliers, including Applied Biosystems. Any other means for this synthesis may also be employed, and the current synthesis of oligonucleotides is well known to those skilled in the art. It is also well known to use similar techniques for preparation. Other oligonucleotides such as thioacids S and alkylated derivatives are well known in the art using similar techniques and commercially available modified DNA synthetic amidite and fixed pore glass (CPG) products (eg, Fluorescently labeled, biotinylated by biosynthetic, luciferin, acridine or psoralen modified DNA synthesis nucleotides and / or CPG (available from Glen Research, Sterling, VA) Or other modifications Nucleotides, such as cholesterol-modified oligonucleotides. According to the present invention, the use of modifications (e.g., using LNA monomers) to enhance the efficacy, specificity, and duration of action and broaden the administration of oligonucleotides includes, for example, Chemical methods such as MOE, ANA, FANA, PS, etc. This can be achieved by using LNA monomers instead of some of the monomers in the current oligonucleotide. LNA-modified oligonucleotides can have dimensions similar to the parent compound or Preferably, the LNA-modified oligonucleotide has less than about 70%, more preferably less than about 60%, most preferably less than about 50% of the LNA monomer, and the size thereof Between about 5 and 25 nucleotides, more preferably about 12 and 20 cores 156342.doc • 54·201143782. Preferred modified oligonucleotide backbones include, but are not limited to, phosphorothioates, palmitic phosphorothioates, dithiophosphates, phosphotriesters, aminoalkyl phosphates, methyl phosphonates, and Other alkyl phosphonates (including phosphonic acid 3-alkyl esters and palmitic phosphonates), phosphinates, amino phosphates (including amine-based acid 3-amino esters and amine-based amines) Alkyl ester), thiocarbonylamine-based acid-decomposing S, phosphonic acid sulfur-based sulphuric acid vinegar, sulfur-based ketone-based linoleic acid triacetate, and a common 3'-5' linkage 2, _5' connection analogs of these main chains, C3 and those with opposite polarity (where the connection of adjacent nuclear unit pairs becomes 5'-3' or 2'-5' becomes 5 '-2'). Also included are various salts, mixed salts and free acid forms. Representative U.S. patents including the above-described broken bonds include, but are not limited to, U.S. Patent Nos. 3,687,808, 4,469,863, 4,476,301, 5,023,243, 5,177,196, 5,188,897, 5,264,423, 5,276,019, 5,278,302, Q 5,286,717, 5,321,131, 5,399,676, 5,405,939, 5,453,496, 5,455,233, 5,466,677, 5,476,925, 5,519,126, 5,536,821 No. 5,541,306, 5,55, η, pp. 5,563,253, 5,571, 799, 5,587, 361, and 5, 625, 050, each incorporated herein by reference. Preferred modified nucleotide backbones containing no phosphorus atoms have a backbone formed by the following: short chain alkyl or cycloalkyl nucleoside linkages, mixed heteroatoms, and alkyl or cycloalkyl nucleosides Inter-bonding, or one or more short-bond heteroatoms or heterocycles 156342.doc -55- 201143782 Internucleoside linkages. These main chains include those of ώ ώ 寺 具有 吗 ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( ( The main chain of the methyl ketone group; the main chain of the methylene ketone and the thioformamyl group; the main chain containing the oxime 3 with an olefin; the main chain of the amino sulfonic acid s; Amine and methylene, its f-base group main chain; sulfonate and sulfonate, the main bond of the amine, and the main bond of the amine; and: i: he and the ancient.. 3⁄4 people Χτ, he has mixed Ν, 〇, S and Representative of CH2. Representative US patents for cultivating and nucleating nucleosides include, but are not limited to, U.S. Patents 5'〇34,506, 5,166,315, 5,185,444, 5,214,134, 帛5,216,141 No. 5, 235, 〇 33, 5'264, 562, 5, 264, 564 胄, 帛 5, 4 〇 5, 938, 5, 434, 257, 5, partial, 677 胄, 帛 5, 47 〇, 967, 5 '489'677, 帛5,541,307, 帛5,561,225, 5,596,086, 5 602 240 胄, 帛5 61〇289, 5'602'240, 5,608,04·6 1帛5, 610, 289, 5, 618 '704, 5, 623, 〇 7 〇胄, 5, 663, 312, 5, 633, 360, 5, 677, 437, and 5, 677, 439, each of which is incorporated herein by reference. In the nucleotide mimetic, both the sugar and the internucleosis linkage (ie, the backbone) in the nucleotide unit are replaced by a new group. The base unit is retained to hybridize with a suitable nucleic acid target compound. A poly compound (a nucleotide mimetic that has been shown to have excellent hybridization properties) is called a peptide nucleic acid (ΡΝΑ). In a ruthenium compound, the sugar-primary bond in the nucleotide is contained in the guanamine-containing backbone, specifically The aminoethylglycine backbone is substituted. The nucleobase is retained and directly or indirectly bonded to the aza nitrogen atom of the guanamine moiety of the backbone 156342.doc-56-201143782. A representative US representative of the preparation of PNA compounds Patents include, but are not limited to, U.S. Patent Nos. 5,539,082, 5, 714, 331, and 5, 719, 262, each of which is incorporated herein by reference. 4, 1497-1500. An embodiment of the invention is an oligonucleotide having a phosphorothioate backbone and a nucleoside having a hetero atom backbone, and in particular, the above-mentioned U.S. Patent No. 5,489,677 -CH2-NH-0-CH2-, -CH2-N(CH3)-0-CH2-[called methylene (methylimido) or MMI backbone], -CH2-0-N ( CH3)-CH2- '-CH2N(CH3)-N(CH3)CH2- and -0-N(CH3)-CH2-CH2-[where the natural phosphodiester backbone is represented as -0-P-0-CH2- And the guanamine backbone of U.S. Patent No. 5,602,240, incorporated herein by reference. Also preferred are the oligonucleotides having the morpholino backbone structure of U.S. Patent No. 5,034,506, the disclosure of which is incorporated herein by reference. The modified oligonucleotide may also contain one or more substituted sugar moieties. Preferred oligonucleotides include one of the following in the 2' position: OH; F; 0-, S- or N-alkyl, Ο-, S- or N-alkenyl; 0-, S- or N - alkynyl; or decyl-fluorenyl-alkyl, wherein the alkyl, alkenyl and alkynyl groups may be substituted or unsubstituted C1 to C6 alkyl or C2 to C10 alkenyl and alkynyl groups. Particularly preferred are 〇(CH2)n OmCH3, 0(CH2)n, OCH3, 0(CH2)nNH2, 0(CH2)nCH3, 0(CH2)n0NH2, and 〇(CH2nON(CH2)nCH3)2, where n And m can be from 1 to about 10. Other preferred oligonucleotides include one of the following in the 2' position: a C1 to C6 lower alkyl 'substituted lower alkyl, alkaryl, aralkyl, fluorenyl-aryl or 0-aryl Alkyl, SH, SCH3, OCN, C, Br, CN, 156342.doc -57- 201143782 CF3, OCF3, SOCH3, S02CH3, 0N02, N02, N3, NH2, heterocycloalkyl, heterocycloalkylaryl, amine Alkylamino group, polyalkylamine group, substituted alkylene group, RNA cleavage group, acceptor group, intercalator, group of pharmacokinetic properties of modified oligonucleotide, or modified oligo A group of pharmacodynamic properties of a glycoside, and other substituents having similar properties. Preferred modifications include 2'-methoxyethoxyethoxy (2'-0-CH2CH20CH3, also known as 2,-0-(2-methoxyethyl) or 2'-MOE), ie, alkoxy Alkoxy group. Other preferred modifications include 2'-didecylaminooxyethoxy (i.e., 〇(CH2)20N(CH3)2 group, also known as T-DMAOE, as described in the Examples below), And 2'-didecylaminoethoxyethoxy (also known in the art as 2'-0-didecylaminoethoxyethyl or 2'-DMAEOE), ie, 2'-0 -CH2-0-CH2-N(CH2)2). Other preferred modifications include 2'-decyloxy (2'-0-CH3), 2'-aminopropoxy (2'-OCH2CH2CH2NH2) and 2'-fluoro (2'-F). Similar modifications can be made at other positions on the oligonucleotide, especially at the 3' end nucleotide or the 2'-5' ligation oligonucleotide in the 3' and 5' end nucleotides of the sugar. 'Bit. The oligonucleotide may also have a sugar mimetic, for example, using a cyclobutyl moiety in place of the pentofuranosyl sugar. Representative U.S. patents which teach the preparation of such modified sugar structures include, but are not limited to, U.S. Patent Nos. 4,981,957, 5,118,800, 5,319,080, 5,359,044, 5,393,878, 5,446,137, 5,466,786, Nos. 5,514,785, 5,519,134, 5,567,811, 5,576,427, 5,591,722, 5,597,909, 5,610,300, 5,627,053, 5,639,873, 5,646,265, 156,342.doc •58 - 201143782 5,658,873, 5,670,633, and 5,700,920 each incorporated herein by reference. Oligonucleotides can also be modified or replaced by a verification base (often abbreviated as "test base" in the industry). As used herein, "unmodified" or "natural" nucleotides include purine bases (adenine (A) and guanine and pyrimidine bases (thymine (T), beetle bite (C), and urinary bites ( U)). Modified nucleotides include other synthetic and natural nucleotides, such as 5-methylcytosine (5-me_C), 5-hydroxymethyl beetle, yellow noise, hypoxanthine, 2 - Amino adenine, glandular sputum and 6-methyl and other alkyl derivatives of guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2 thiopyrimidine, 2-thiothymidine and 2-thiocytosine, 5-protonated uracil and cytosine, 5-propynyl uracil and cytosine, 6-oxyuracil, cytosine and thymine, 5- Uracil (false urinary sputum), 4-thiourine bite, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-carbyl and other 8-substituted adenine And guanine, 5-halogen, especially 5--; odor, 5-trifluoromethyl and other 5-substituted urinary mouth bites, 7-mercapto guanine and 7-mercapto adenine, 8 -azaguanine and 8-azadenine, 7-deazaguanine and 7-deaza adenine and 3- Denitrified guanine and 3 - deaza adenine. In addition, the nucleotides include those disclosed in U.S. Patent No. 3,687,808, the disclosure of which is incorporated herein by reference to the " The Concise Encyclopedia of Polymer Science And Engineering, pp. 858-859 Page, Kroschwitz, JI Editor, John Wiley & Sons, 1990, by Englisch et al., "Angewandle Chemie, International Edition", 1991, 30, p. 613, and by Sanghvi, YS, Chapter 15, 156342.doc -59-201143782 "Antisense Research and Applications", pp. 289-302, Crooke, ST and Lebleu, B. ea., CRC Press, 1993. Some of these nucleotides are especially useful. To increase the binding affinity of the oligomeric compound of the present invention. The nucleotides include a 5-substituted mouth bite, a 6-aza. The bite and N-2, Ν-6 and 0-6 substituted 嗓吟' include 2- Aminopropyl adenine, 5-propionyl uracil and 5-propynyl cytosine. 5-methylcytosine substitution has been shown to increase the stability of nucleic acid duplexes by 0.6-1.2 ° C (Sanghvi, YS, Crooke, ST and Lebleu, B. Editor, "Antisense Research and A Pplications, CRC Press, Boca Raton, 1993, pp. 276-278) and are currently preferred base substitutions, even more particularly when combined with 2'-methoxyethyl saccharide modifications. Representative U.S. patents which teach the preparation of the above-described modified nucleotide acid and other modified nucleic acids include, but are not limited to, U.S. Patents 3,687,8-8, and 4,845,205, 5,130,302, 5,134,066. , Nos. 5,175,273, 5,367,066, 5,432,272, 5,457,187, 5,459,255, 5,484,9-8, 5,502,177, 5,525,711, 5,552,54, 5,587,469 No. 5,596,091, 5,614,617, 5,750,692, and 5,681,941 each incorporated herein by reference. Another modification to the oligonucleotides of the invention involves chemically linking one or more conjugates that enhance the activity, cellular distribution or cellular uptake of the oligonucleotide to the nucleotide. *Hai and other parts include, but are not limited to, lipid parts (such as cholesterol part), biliary 156342.doc • 60-201143782 hexyl-s-triphenylmercaptothiol), thiocholesterol, lipodiene diacetate or undecane Base residues), phospholipids (eg, acid, sulfur scales (eg, aliphatic bonds (eg, dodeca-hexadecyl-racemic-glycerol or triethylammonium 1,2,di-anthracene-hexa) Alkyl--racemic-glycerol-3-H-phosphonic acid vinegar, polyamine or polyethylene glycol chain, or diamondine acetic acid, palmitoyl moiety, or octadecylamine or hexylamino-carbonyl-oxycholesterol Representative US patents that teach the preparation of such oligonucleotide primers include Ο

However, it is not limited to US 5,218,105, 5.545.730, 5.580.731, 5,109,124, 5,414,077, 5,578,718, 4,605,735, 4,789,737, 4,876,335, 5.082.830, 5.082.830 , 5, 245, 022, 5, 262, 536, 5, 317, 098, 5, 416, 203, 5, 512, 667, Bliss 4, 828, 979, 5, 525, 465, 5, 552, 538, 5, 580, 731, 5, 118, 802, 5, 486, 603, pp. 5,608,046, 4,667,025, 4,824,941, 4,904,582, 5,112,963, 5,112,963, 5,254,469, 5,272,250, 5,371,241, 5,451,463 No. 5,514,785, 4,948,882, 5,541,313, 5,578,717, 5,591,584, 5,138,045, 5,512,439, 4,587,044, 4,762,779, 4,835,263, 4,958,013, 5.214.136, 5.214.136, 5,258,506, 5,292,873, 5, 391,723, 5,510,475, 5,565,552, 156,342.doc-61-201143782 5,567,810, 5,574,142, 5,585,481, 5,587,371, 5,595,726, 5,597,696, 5,599,923, Nos. 5, 599, 928, and 5,688, 941 each incorporated herein by reference. Drug Research. The compounds of the invention may also be added to the area of drug research and dry validation. The present invention encompasses the use of the compounds identified herein and preferred target segments for drug research, which attempts to elucidate the relationship between a UM homologous sequence 2 (LHX2) polynucleotide and a disease state, phenotype, or condition. The relationship of existence. The methods comprise detecting or modulating an LHX2 polynucleotide comprising contacting a sample, tissue, cell, or organism with a compound of the invention, and measuring the nucleic acid or protein of the LHX2 polynucleotide at a time after treatment. And/or related phenotype or chemically untreated sample or another sample in the present invention' and a comparatively measured value and compound treated sample as needed. The methods may also be performed in parallel or in combination with other experiments to determine the function of an unknown gene for the t-validation process' or to determine a particular base@product for use in treating or preventing a particular disease, condition, or phenotype (4) Effectiveness. Evaluation of gene expression up-regulation or inhibition: The transfer of exogenous nucleic acid into a host cell or organism can be assessed by directly detecting the presence of nucleic acid in the cell or organism. This detection can be achieved by a number of methods well known in the art. For example, the presence of an exogenous nucleic acid can be detected by Southern blotting or by polymerase chain reaction (better technique for the amplification of a nucleic acid-related nucleotide sequence by a heterosexual amplification). Conventional methods for measuring the performance of exogenous nucleic acids. For example, Northern blot and reverse transcription pcR (RT_pcR) can be used to detect mRNA produced from an exogenous nucleic acid by J56342.doc-62-201143782. Activity or reporter protein activity to detect the expression of RNA in an exogenous nucleic acid. For example, the antisense modulating activity can be indirectly measured according to the decrease or increase in the performance of the target nucleic acid, thereby indicating that the exogenous nucleic acid is in the effector-subRNA Based on sequence conservation, primers can be designed and used to amplify the coding region of the target gene. Initially, the highest expression coding region from each gene can be used to create a control gene model, but any coding or non-coding region can be used. Each control gene is assembled by inserting each coding region between the reporter coding region and its r\^P〇ly(A) signal. These plasmids will be generated on the gene. The swim portion has a reporter gene and has a potential RNAi target mRNA in the 3' non-coding region. The effectiveness of the individual antisense oligonucleotides is analyzed by modulating the reporter gene. Reporter genes for use in the methods of the invention Containing acetaminolate synthase (AHAS), alkaline phosphatase (AP), beta galactosidase (LacZ), beta glucuronidase (GUS), chloramphenicol acetyltransferase (CAT), Green fluorescent protein (GFP), red fluorescent protein (RFP), yellow ^ fluorescent protein (YFP), cyan fluorescent protein (CFP), horseradish peroxidase (HRP), luciferase (Luc) , nopaline synthase (NOS), octopine carnitine synthase (OCS), and its derivatives. A variety of markers can be used to confer resistance to: • Selectable markers: ampicillin (ampicillin), bleomycin ( Bleomycin), chloramphenicol, gentamycin, hygromycin, kanamycin, lincomycin, methotrexate, grass Phosphothosicin, puromycin, and tetracycline. Determination of reporters Gene 156342.doc -63- 201143782 Methods of modulation are well known in the art and include, but are not limited to, fluorescent methods (eg, fluorescence spectroscopy, fluorescence activated cell sorting (FACS), fluorescence microscopy), Antibiotic Resistance Assays LHX2 protein and mRNA expression can be analyzed using methods known to those skilled in the art and described elsewhere herein. For example, immunological assays such as ELISA can be used to measure protein content. The lhx2 ELISA assay kit is commercially available', for example, from R&D Systeins (Minneap® Us, MN). In the examples, the sample treated with the antisense oligonucleotide of the present invention (for example, cells or tissues in vivo or in vitro) is evaluated by comparison with the Lhx2 expression in the control sample. LHX2 behaves (eg, mRNA or protein). For example, methods known to those skilled in the art can be used to compare the performance of a protein or nucleic acid to a simulated or untreated sample. Alternatively, the treated sample can be compared to a control antisense oligonucleotide (e.g., with altered or different sequences) depending on the desired information. In another embodiment, the treated sample and the untreated test can be compared using differences in the performance of different nucleic acids (including any criteria deemed suitable by the investigator, eg, housekeeping genes) in the treated and untreated samples. The difference in the expression of LHX2 protein or nucleic acid in the sample. The observed difference can be expressed, for example, in ratios or parts as needed for comparison with a control group. In an embodiment, the amount of LHX2 mRNA or protein in the sample treated with the antisense nucleus acid of the present invention is increased or decreased by about 1.25 times to about 1 with respect to the untreated sample or the sample treated with the control nucleic acid. 〇 times or higher. In embodiments, the LHX2 mRNA or egg 156342.doc-64-201143782 white matter is increased or decreased by at least about 1.25 times, at least about 1.3 times, at least about 1.4 times, at least about 1.5 times, at least about 1.6 times, at least about 1.7.倍, at least about 1.8 times, at least about 2 times, at least about 2.5 times, at least about 3 times, at least about 3.5 times, at least about 4 times, at least about 4.5 times, at least about 5 times, at least about 5.5 times, at least about 6 Multiplier, at least about 6.5 times, at least about 7 times, at least about 7.5 times, at least about 8 times, at least about 8.5 times 'at least about 9 times, at least about 9.5 times, or at least about 10 times or more.

Kits, Research Reagents, Diagnosis, and Therapy The compounds of the present invention are useful in the diagnosis, treatment, and prevention, and are useful as kits for research reagents and components. In addition, those skilled in the art typically employ antisense nucleotides that are capable of inhibiting gene expression with strong specificity to elucidate the function of a particular gene or to distinguish the function of each member of the biological pathway. For use in kits and diagnostics and in a variety of biological systems, the present sigma sigma (alone or in combination with other compounds or therapeutic agents) can be used as a tool in differential and/or combinatorial analysis to elucidate cells and The pattern of expression of a portion of the gene expressed in the tissue or the entire complementary sequence. The term "biological system" or "system" as used herein is defined to mean any organism, cell, cell culture or tissue that exhibits, or is sufficient to represent, the LIM homolog 2 (LHX2) gene product. Such systems include, but are not limited to, classes, transgenic animals, cells, cell cultures, & plants, grafts, and combinations thereof.移植 移植 理 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , Analyze the resulting pattern for the degree of difference in genetic performance of 156342.doc -65- 201143782 correlation, signaling pathway, cell localization, degree of performance, size, structure or function of the gene being tested. Such assays can be performed in stimulated or unstimulated cells and in the presence or absence of other compounds that affect performance patterns. Examples of methods for gene expression analysis known in the art include DNA arrays or microarrays, SAGE (series analysis of gene expression), READS (digestion amplification of digestive (7), TOGA (general gene expression analysis), protein arrays and Proteomics, sequenced display (EST) sequencing, subtractive RNA fingerprinting (Surf), subtractive selection, differential display (DD), comparative genomic hybridization, FISH (fluorescence in situ hybridization) and mass spectrometry. The compounds of the invention are useful in research and diagnostics because such compounds can hybridize to nucleic acids encoding LIM homolog 2 (LHX2). For example, oligonucleotides that hybridize under the efficiency and conditions of an effective LHX2 modulator as disclosed herein are effective primers or probes and probes under conditions that facilitate gene amplification or detection, respectively. Needles can be used in methods that require specific detection of nucleic acid molecules encoding LHX2 and amplification of such nucleic acid molecules for detection or for use in other LHX2 studies. Hybridization of the antisense oligonucleotide (especially primers and probes) of the present invention with a nucleic acid encoding LHX2 can be detected by means of f. Such means may include coupling the enzyme to the nucleus (4), a radiolabeled oligonucleotide, or any other suitable means of detection. It is also possible to manufacture a kit that detects the (3) phantom content in the sample using an edge detection method. Those skilled in the art also utilize the specificity and sensitivity of antisense compounds in therapeutic applications. Antisense compounds have been used as therapeutic components to treat disease states in animals (3 humans). Anti-Stupidity Hanyi Nutrient Music is safe and effective 156342.doc -66 - 201143782 The field is being administered to humans and many clinical trials are currently being implemented. It is thus determined that the antisense compound can be a useful therapeutic modality that can be used in the treatment of cells, tissues, and animals, particularly humans. At the time of treatment, an animal (preferably a human) suspected of having a disease or condition treatable by modulating the expression of LHX2 polymorphic acid is treated by administering an antisense compound of the present invention. For example, in one non-limiting embodiment, the methods comprise the step of administering to a subject in need of treatment a therapeutically effective amount of an LHX2 modulator. The LHX2 modulator of the present invention is effective for regulating the activity of LHX2 or regulating the performance of LHX2 protein. In one embodiment, the activity or performance of LHX2 in the animal is inhibited by about 10% compared to the control. Preferably, the activity or performance of LHX2 in the animal is inhibited by about 30%. More preferably, the activity or performance of LHX2 in the animal is inhibited by 50% or more. Thus, an oligomeric compound can modulate the expression of LIM homolog 2 (LHX2) mRNA by at least 10%, at least 50%, at least 25%, at least 30%, at least 40%, at least 50%, at least as compared to a control group. 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%. In one embodiment, the activity or performance of LIM homolog 2 (LHX2) in an animal is increased by about 10% compared to a control group. Preferably, the activity or performance of LHX2 in the animal is increased by about 30%. More preferably, the activity or performance of LHX2 in animals is increased by 50% or more. Thus, the oligomeric compound can modulate the performance of the LHX2 mRNA by at least 10%, at least 50%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, compared to the control group. At least 75%, at least 80%, at least 156,342.doc -67.201143782 85/〇, at least 90%, at least 95%, at least 98. /. At least 99%, or 100%. For example, the reduction in the expression of UM homologous sequence 2 (LHX2) in serum, blood, adipose tissue, liver or any other body fluid, tissue or organ of an animal can be measured. Preferably, the cells contained in the fluid, tissue or organ analyzed contain nucleic acid molecules encoding the LHX2 peptide and/or the LHX2 protein itself. The compound can be used in a pharmaceutical composition by adding an effective amount of a compound of the invention to a suitable pharmaceutically acceptable diluent or carrier. The uses and methods of the compounds of the invention may also be used for prophylactic purposes. Conjugates Another modification of the nucleotides of the present invention involves the chemical attachment of one or more moieties or conjugates that enhance the activity, cellular distribution or cellular uptake of the nucleotide to the nucleotide. The moieties or conjugates may comprise a coupling group covalently bonded to a functional group such as a primary or secondary hydroxyl group. The coupling group of the present invention comprises an intercalating agent, a reporter molecule, a polyamine, a polyamine, a polyethylene glycol, a polymymy, a group for enhancing the pharmacodynamic properties of the polymer, and an oligo reinforced group. a group of pharmacokinetic properties f. Typical coupling groups include cholesterol, lipids, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, luciferin, rh〇damine, coumarin, and dyes. . In the context of the present invention, a group that enhances pharmacodynamic properties comprises a group that improves uptake, enhances degradation resistance, and/or enhances sequence-specific hybridization with dry nucleic acids. In the context of the present invention, the group which enhances the metabolic kinetics of the drug H contains <RTI ID=0.0>>>>> A representative coupling group is disclosed in International Patent Application No. PCT/US92/09196, filed on Oct. 23, 1992, and hereby incorporated herein by reference. Coupling moieties include, but are not limited to, lipid moieties (eg, cholesterol moieties), cholic acid, thioethers (eg, hexyl-5-trityl mercaptan), thiocholesterol, aliphatic chains (eg, dodecanediol) Or undecyl residue), phospholipid (for example, di-hexadecyl-racemic-glycerol or triethylammonium 1,2-di-indole-hexadecyl-racemic-glycerol_3_H Phosphonate), polyamine or polyethylene glycol chain, or adamantane acetic acid, palmitoyl moiety, or octadecylamine or hexylamino-carbonyl-oxycholesterol moiety To active drug substances 'eg aspirin, warfarin, phenylbutazone, ibuprofen, suprofen, fenbufen, ketoprofen ), (S)-(+)-pranoprofen ((S)-(+)-pranoprofen), carprofen, dansylsarcosine, 2,3,5-triiodobenzene Sulfonic acid, flufenamic acid, leucovorin, benzophenone, chlorinated D-Qin, diazep, ° D-omeemine (indomethicin), barbiturate, Cephalosporin, a tranexamine, an antidiabetic, an antibacterial or an antibiotic. Representative U.S. patents which teach the preparation of such nucleotide conjugates include, but are not limited to, U.S. Patent Nos. 4,828,979, 4,948,882, 5,218,105, 5,525,465, 5,541,313, 5.545. 730, 5, 552, 538, 5, 578, 717, 5.580.731, 5, 580, 731, 5, 591, 584, 156, 342. doc - 69 - 201143782 5, 109, 124, 5, 118, 802, 5, 138, 045, 5, 414, 077, 5, 486, 603, , No. 5, 512, 439, No. 5, No. 5, 598, pp. No. 5, 082, 013, No. 5, 082, 830, No. 5, 112, 963, No. 5, 214, 136, No. 5, 082, 830, No. 5,112, 963, No. 5, 214, 136, No. 5, 245, 022, No. 5, 258, 506, No. 5, 258, 506, No. 5, 262, 536, No. 5, 272, 250, No. 5, 292, 873 No. 5,317,098% Nos. 5, 371, 241, 5, 416, 203, 5, 451, 463, 5, 510, 475, 5, 512, 667, 5, 514, 785, 5, 565, 552, 5, 567, 810 & 5, 574, 142, 5, 585, 481, Nos. 5,587,371 & 5,595,726, 5,597,696, 5,599,923, 5,599,928 and 5,688,941. Formulations The compounds of the invention may also be mixed, encapsulated, coupled or otherwise with other molecules, molecular structures or mixtures of compounds (eg, liposomes, molecules that target receptors, oral, rectal, topical or other formulations). The combination is combined to aid in ingestion, distribution and/or absorption. Representative U.S. patents which teach the preparation of such formulations which facilitate ingestion, distribution and/or absorption include, but are not limited to, U.S. Patent Nos. 5,108,921, 5,354,844, 5,416,016, 5,459,127, 5,521,291. '5, 543, 165, 156 342. doc • 70 · 201143782 5, 547, 932, 5, 583, 020, 5, 591, 721, 4, 426, 330, 4, 534, 899, 5, 013, 556, 5, 108, 921, 5, 213, 804, Nos. 5,227,170, 5,264,221, 5,356,633, 5,395,619, 5,416,016, 5,417,978, 5,462,854, 5,469,854, 5,512,295, 5,527,528, 5,534,259, 5,543,152 No. 5,556,948, 5,580, 5 <75, and 5,595,756 each incorporated herein by reference. Although it is not necessary to administer antisense nucleoside acid in the context of the vector in order to modulate performance and/or function, embodiments of the invention relate to expression vector constructs for expression of antisense oligonucleotides, including promoters , a hybrid promoter gene sequence and having a strong constitutive promoter activity, or a promoter activity that can be induced under the desired conditions. In one embodiment, the practice of the invention involves the administration of at least one of the above antisense oligonucleotides using a suitable nucleic acid delivery system Q. In one embodiment, the system comprises a non-viral vector operably linked to a polynucleotide. Examples of such non-viral vectors include only oligonucleotides (e.g., any one or more of SEq ID NO: 5 to 16) or in combination with a suitable protein, polysaccharide or lipid formulation. Further, a suitable nucleic acid delivery system comprises a viral vector, typically from adenosis, adeno-associated virus (AAV), helper cell-dependent adenosis, retrovirus, or Japanese liposome-hemagglutinating virus (HVJ) complex. A sequence of at least one of them. Preferably, the viral vector comprises a strong eukaryotic promoter (e.g., a cytomegalovirus (CMV) promoter) operably linked to 156342.doc-71-201143782 to a polynucleotide. Further preferred carriers comprise viral vectors, fusion proteins and chemical conjugates. The retroviral vector comprises Moloney mouse leukemia virus and HIV-based virus. A preferred HIV-based viral vector comprises at least two vectors, wherein the gag and pol genes are from the HIV genome and the env gene is from another virus. The dnA disease vector is preferred. Such vectors comprise a ρ〇χ vector (e.g., orthopoxvirus or acne virus vector), a herpesvirus vector (e.g., herpes simplex prion (HSV) vector), an adenoviral vector, and an adeno-associated viral vector. A compound of the invention encompasses any pharmaceutically acceptable salt, ester, or salt of such an ester, or is capable of providing (directly or indirectly) a biologically active metabolite or residue thereof after administration to an animal (including humans) Any other compound. The term "pharmaceutically acceptable salt" refers to a physiologically and pharmaceutically acceptable salt of a compound of the present invention: that is, a salt which retains the desired biological activity of the parent compound and which does not impart an undesirable toxicological effect. For oligonucleosides, the preferred examples of the salts of the T-accepting salts and their use are further described in U.S. Patent No. 6,287,860, incorporated herein by reference. The invention also encompasses pharmaceutical compositions and formulations comprising the antisense compounds of the invention. The pharmaceutical compositions of the present invention can be administered in a variety of ways, depending on the desired local or systemic treatment and the area to be treated. Administration may be local (including ocular administration and administration (4), including vaginal and rectal delivery), lung (eg, by inhaling or spraying powder or aerosol, including by atomizer); , intranasal, epidermal and percutaneous, oral or parenteral administration. Parenteral 156342.doc -72- 201143782 administration includes intravenous, intra-arterial, subcutaneous, intraperitoneal or intramuscular injection or infusion; or intracranial (eg, intrathecal or intraventricular) administration. For treatment of tissue in the central nervous system, administration can be by, for example, injection or infusion into the cerebrospinal fluid. The administration of antisense RNA in cerebrospinal fluid is described, for example, in "Methods for slowing familial ALS disease progression", U.S. Patent Application Publication No. 2007/0117772, the entire contents of which is incorporated herein by reference. When it is intended to administer an antisense oligonucleotide of the present invention to a cell of the central nervous system, one or more agents capable of promoting penetration of the subject antisense oligonucleotide through the blood-brain barrier may be administered. Injections can be made, for example, in the entorhinal cortex or hippocampus. The delivery of a neurotrophic factor by administering an adenoviral vector to a motor neuron in muscle tissue is described, for example, in "Adenoviral-vector-mediated gene transfer into medullary motor neurons", U.S. Patent No. 6,632,427, incorporated herein by reference. Incorporated herein. It is known in the art to deliver a carrier directly to the brain (e.g., striatum, thalamus, hippocampus, or substantia nigra) and is described in, for example, U.S. Patent No. 6,756,523, "Adenovirus vectors for the transfer of foreign genes into cells". In the central nervous system particularly in brain", which is incorporated herein by reference. Administration can be by rapid administration by injection or by slow infusion or administration of a sustained release formulation over a period of time. The title antisense oligonucleotide can also be linked or coupled to an agent that provides the desired pharmaceutical or pharmacodynamic properties. For example, an antisense oligonucleotide can be coupled to any substance known in the art to promote permeation or delivery in the blood-brain barrier (eg, an antibody to the iron-protein 156342.doc-73-201143782 transfer protein receptor), and It is administered by intravenous injection. The antisense compound can be linked to a viral vector which can, for example, render the antisense compound more effective and/or increase the delivery of the antisense compound in the blood-brain barrier. The osmotic blood brain barrier can also be destroyed by, for example, infusion of sugars including, but not limited to, meso-erythritol, xylitol, D(+) galactose, D(+) lactose, D (+) xylose, dulcitol, muscle-inositol, L(-) fructose, D(-) mannitol, D(+) glucose, D(+) arabinose, D(-) arabinose, fiber two Sugar, D(+) maltose, D(+) raffinose, L(+) rhamnose, D(+) melibiose, D(-) ribose, ribitol, D(+) arabitol, L (-) arabitol, D(+) fucose, L(-) fucose, D (one) to threose, L(+) to threose, and L(-) tosuose; or amino acid, Including but not limited to glutamine, lysine, arginine, aspartame, aspartic acid, cysteine, glutamic acid, glycine, histidine, leucine, methyl sulfide Amine acid, phenylalanine, valine, serine, threonine, tyrosine, proline, and taurine. Methods and materials for enhancing blood-brain barrier penetration are described, for example, in "Method for the delivery of genetic material across the blood brain barrier", No. 6,294,042, "Material for passage through the blood-brain". "Blocker" and "Parotheral delivery systems", No. 6,936,589, the entire contents of each of which are incorporated herein by reference. The title antisense compound can be mixed, encapsulated, coupled or otherwise combined with other molecules, molecular structures or mixtures of compounds (eg, liposomes, molecules that target receptors, oral, rectal, topical or other formulations). To help ingest, distribute and / or absorb. For example, cationic lipids may be included in the formulation 156342.doc-74-201143782 to promote oligonucleoic acid uptake. One of the compositions shown to promote capture is LIPOFECTIN (available from GIBCO-BRL, Bethesda, MD). It is believed that at least one 2'-0-methoxyethyl modified nucleotide is particularly useful for oral administration. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders. Pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be required or desired. You can also use a wrap 避孕 condom, gloves, and the like. The pharmaceutical formulations of the present invention which may be conveniently presented in unit dosage form can be prepared according to conventional techniques well known in the pharmaceutical industry. Such techniques include the step of bringing the active ingredient into association with a pharmaceutical carrier or excipient. In general, the formulations may be prepared by uniformly and intimately bringing the active ingredient into the liquid carrier or the fine solid carrier or both, and then, if necessary, shaping the product. The compositions of the present invention may be formulated into any of a number of possible dosage forms, such as, but not limited to, lozenges, 'knee pockets, gel capsules, liquid syrups, soft gels, suppositories, and enemas. The compositions of the present invention may also be formulated as suspensions in aqueous, non-aqueous or mixed media. The aqueous suspension may further contain a substance which increases the viscosity of the suspension, and contains, for example, sodium carboxymethyl cellulose, sorbitol and/or dextran. The suspension may also contain a stabilizer. The pharmaceutical compositions of the present invention include, but are not limited to, solutions, emulsions, foams, and formulations containing liposomes. The pharmaceutical compositions and formulations of the present invention may include one or more penetration enhancers, carriers, excipients or other active or inert ingredients of 156342.doc • 75- 201143782. 2 (4) (4) Dispersed in another liquid in the form of droplets (diameter through a heterogeneous system. The emulsion may contain additional components in addition to the dispersed phase, and may be present as a solution in the secondary phase, sub 7 aqueous phase, oil phase or itself As an active drug for the separation phase, the present application contains a microemulsion. The emulsion and its use are well known in the art and are described in U.S. Patent No. 6,287,860. The formulation of the present invention comprises a lipid. The term "liposome" as used in the present invention means a vesicle composed of two amphiphilic lipids in one or one spherical double layer. A lipid system monolayer or multilamellar vesicle a membrane having a self-lipophilic! raw material formation and an aqueous internal structure containing the composition to be delivered. The positively charged liposomes of the cationic oligosaccharide system are believed to interact with negatively charged ship molecules to form a stable complex. Liposomes that are pH-sensitive or negatively charged are believed to be captureable rather than complexed. Both ionic and non-cationic liposomes can be used to deliver DNA to cells. $Fat (4) also contains "spatial-stable" lipids. (d), the technique used herein." A button comprising one or more specific lipids. In the inclusion of liposomes, such characteristic lipids produce enhanced cycle life relative to liposomes lacking the particular lipids. Liposomes. Examples of sterically stabilized liposomes are those in which the vesicle-forming lipid portion of the liposome comprises one or more glycolipids or a hydrophilic or polymeric polymer (eg, polyethylene glycol (peg) moiety). Liposomes and their use are further described in U.S. Patent No. 6,287,860. The pharmaceutical formulations and compositions of this month may also contain surfactants. Table 156342.doc -76- 201143782 Surfactants in pharmaceutical products, The use of formulations and emulsions is well known in the art. Surfactants and their use are further described in U.S. Patent No. 6,287,860, which is incorporated herein by reference in its entirety herein in Agent to achieve effective delivery of nuclear 'acids, especially nucleotides. In addition to contributing to the diffusion of non-lipophilic drugs in the cell membrane, penetration enhancers also enhance lipophilic drugs Permeability of substances. Penetration enhancers can be classified into one of five categories: that is, surface active agents, fatty acids, bile salts, chelating agents, and non-chelating non-surfactants ° penetration enhancers and Its use is further described in U.S. Patent No. 6,287,860, the disclosure of which is incorporated herein by reference in its entirety in its entirety in the the the the the the the the Preferred formulations for topical administration comprise a mixture of the present invention and a local delivery agent, for example, lipids, liposomes, fatty acids, fatty acid esters, steroids, chelating agents, and surface active agents. 〇Preferred lipids and liposomes comprise neutral lipids and liposomes (eg diterpenoids _ phospholipids D〇PE ethanolamine, digestive phosphatidylcholine 1 DMPC, monostearyl phospholipids) Choline choline), negative lipids and liposomes (eg, a ugly phosphatidyl glycerol DMpG) and cationic lipids and lipids (eg, dioleyltetramethylaminopropyl D〇TAp and Two oil base Alcohol amines DOTMA). For topical or other administration, the oligonucleotides of the present invention may be encapsulated in or may form a complex with the liposome (especially cationic liposomes. Alternatively, the nucleophilic acid may be associated with lipids, especially Cationic lipid complexes. 156342.doc -77- 201143782 Preferred fatty acids and esters, pharmaceutically acceptable salts thereof, and uses thereof are further described in U.S. Patent No. 6,287,860. Compositions and formulations for oral administration The composition comprises a powder or a particle, a microparticle, a nanoparticle, a suspension or solution in a water or non-aqueous medium, a capsule, a gel capsule, a sachet, a lozenge or a micro-tablet. It may be desirable to use a thickener. A flavoring agent, a diluent, an emulsifier, a dispersing aid or a binder. Preferred oral formulations are those in which the present invention is administered in combination with one or more penetration enhancers, surfactants, and chelating agents. Preferred surfactants comprise fatty acids and/or esters or salts thereof, bile acids and/or salts thereof. Preferred bile acids/salts and fatty acids and their use are further described in U.S. Patent No. 6,287,86, which is incorporated herein by reference. Reference method incorporated into this Also preferred is a combination of penetration enhancers, for example, a combination of a fatty acid/salt and a bile acid/salt. Particularly preferred combinations are sodium laurate, citric acid and UDCA. Other permeation enhancers include oxyethylene Winter lauric ether, polyoxyethylene, _ cetyl ether. The nucleus acid of the present invention can be orally delivered in the form of particles (including spray dried granules, or composited to form micro or nano granules). The ruthenium complexing agent and its use are further described in U.S. Patent No. 6,287, the disclosure of which is hereby incorporated by reference herein in its entirety herein in I bacteria aqueous solution containing buffers, diluents and other suitable additives, such as, but not limited to, penetration enhancers', carrier compounds and other pharmaceutically acceptable carriers or excipients Certain embodiments of the present invention provide a pharmaceutical group containing one or more oligomeric species and one or more other chemotherapeutic agents that are exerted by a non-antisense mechanism 156342.doc -78· 201143782 Ο

Compound. Examples of such chemotherapeutic agents include, but are not limited to, cancer chemotherapeutic drugs, such as daunorubicin, daunomycin, dactinomycin, doxorubicin, and softness. Epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide, ifosfamide, sputum bite (cytosine arabinoside), bischloroethyl-nitrosurea, busulfan, mitomycin C, actinomycin D, mithramycin, splatter Prednisone, hydroxyprogesterone, testosterone, tamoxifen, dacarbazine, procarbazine, hexamethylmelamine, five Pentamethylmelamine, mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosurea, nitrogen mustar Ds), melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-azacytidine, hydroxyurea, deoxycoformycin, 4-hydroxyperoxycyclo-phosphoramide, 5-fluorourine 5-fluorouracil (5-FU), 5-IL-deoxyuronium (5-FUdR), methotrexate (MTX), colchicines, taxol , Changchun new test (vincristine), Changchun test (vinblastine), 156342.doc -79- 201143782 etoposide (VP-16), trimetrexate (trimetrexate), irinotecan (irinotecan), topotem Topotecan, gemcitabine, teniposide, cisplatin, and diethylstilbestrol (DES). When used with a compound of the invention, the chemotherapeutic agents can be isolated (eg, 5-FU and oligo-acidic acid), sequentially (eg, 5-FU and oligonucleotides, followed by MTX after a certain period of time) And nucleotides), or in combination with one or more other such chemotherapeutic agents (eg, 5-FU, MTX and oligonucleotides, or 5-FU, radiation therapy, and oligonucleotides). Anti-inflammatory drugs (including but not limited to non-steroidal anti-inflammatory drugs and corticosteroids, as well as antiviral drugs, including but not limited to ribivirin, vidarabine, aciclovir (acyci〇v (1) And ganciclovir may also be combined in the compositions of the invention. Combinations of antisense compounds and other non-antisense drugs are also within the scope of the invention. Compounds of two or more combinations may be taken together or sequentially In another related embodiment, the compositions of the present invention may contain one or more antisense compounds (especially oligonucleotides) that target the first nucleic acid and one or more additional antisense compounds that target the second nucleic acid target. For example, the first target may be a specific antisense sequence of the homologous sequence 2 (LHX2), wherein: (4) is a region from another nucleotide sequence. __Select... ^ is selected as the present invention The composition may be: antisense compounds of different regions. Many examples of antisense compounds are disclosed herein and other examples may be selected from suitable compounds known in the art. Compounds of two or more combinations may be used or sequentially Use. Dosing: 156342. Doc 201143782 It is believed that those skilled in the art should be aware of the formulation of therapeutic compositions and their subsequent administration (administration). The administration depends on the severity and responsiveness of the condition to be treated. It lasts for several days to several months, or until the right to cure or reduce the disease state. The optimal drug administration plan can be calculated by measuring the drug accumulation in the patient. Those skilled in the art can easily determine the optimal dosage and dosage method. And the rate of repetition. The optimum dose may vary depending on the relative efficacy of the individual oligonuclei, and can usually be calculated based on the discovery of U(10) in vivo and in vivo (four). In general, the dose is 〇.〇 1 ton / kg body weight to 1 〇〇 g / kg body weight, and can be given daily, weekly, monthly or yearly - one or more times, or even every 2 to 20 years - times. Those skilled in the art The repetition rate and concentration of the drug can be easily estimated based on the measured residence time and concentration of the drug in the body fluid or tissue. After successful treatment, it can be expected that the patient can maintain the therapy to prevent recurrence of the disease state, wherein 'I is 0.01 A maintenance dose between pg/kg body weight and 100 g/k^ weight is administered to raise the bitter acid, and is administered _ times per day to - or more times to every 20 years. In the examples, the following is used Dosage of the drug to treat the patient: at least about 1 mg/kg body weight, at least about 2 mg/kg body weight, at least about 3 mg/kg body weight, to v about 4 mg/kg body weight, at least about 5 mg/kg body weight, at least about 6 Mg/kg body weight to about 7 mg/kg body weight, at least about 8 mg/kg body weight, at least about 9 mg/kg body weight, at least about 10 mg/kg body weight, at least about 15 body weight, to/, spoon 20 mg/kg body weight At least about 25 body weight, at least about mg/kg body weight, at least about 35 mg/kg body weight, at least about 4 mg/kg body weight, to y, spoon 45 mg/kg body weight, at least about 5 mg/kg body weight, at least About (10) 156342.doc

• 8 U 201143782 mg/kg body weight, at least about 7 mg/kg body weight, at least about 8 mg/kg body weight, at least about 90 mg/kg body weight, or at least about 1 inch, body weight. Some of the injectable doses of antisense oligonucleotides are described, for example, in "Antisense m〇dulati〇n 〇fpTpiB up." in U.S. Patent No. 7,563, the entire contents of which is incorporated herein by reference. The various embodiments of the present invention have been described hereinabove, however, it is understood that the embodiments are provided by way of example and not limitation. Many changes may be made to the disclosed embodiments without departing from the spirit and scope of the invention. Therefore, the breadth and scope of the present invention should not be limited by any of the above embodiments. All documents mentioned herein are incorporated herein by reference. All publications and patent documents cited in this application are hereby incorporated by reference in their entirety in the extent of the extent The Applicant does not recognize that any particular reference is a "previous technique" of its invention for each reference cited in this document. Examples of compositions and methods of the invention are illustrated in the following examples. EXAMPLES The following non-limiting examples are illustrative of selected embodiments of the invention. It is to be understood that the ratios of the elements of the components shown and the substitutions thereof are within the scope of the embodiments of the present invention. Example 1: Design of antisense oligonucleotides specific for the antisense nucleic acid molecule of L1M homologous sequence 2 (LHX2) and/or the sense strand of LHX2 polynucleotide, as described above, the term "has "Specific oligonucleotide" or "targeted oligonucleotide" refers to an oligonucleotide having the following sequence: capable of forming a stable complex with one of the target genes, 156342.doc •82-201143782, or (ϋ) is capable of forming a stable duplex with a portion of the mRNA transcript of the targeted gene. Promote the selection of appropriate nucleotides by using a computer program (eg IDT AntiSense Design, IDT OligoAnalyzer) that automatically identifies sub-sequences of i9-25 nucleotides in each given sequence, such The sequence forms a hybrid with the target polynucleotide sequence and has a desired melting temperature (typically 50-60. 〇' and does not form a self-dimer or other complex secondary structure. 〇 by using an auto-alignment nucleic acid sequence and Computer programs indicating areas of homology or homology to further facilitate the selection of appropriate nucleotides. Use of these

The program compares the obtained nucleic acid sequences by, for example, searching a database such as GenBank or by sequencing the PCR products. Comparison of nucleic acid sequences from a range of genes and intergenic regions in a given genome allows for the selection of nucleic acid sequences of the appropriate degree of specificity for the target gene. Such procedures allow for the selection of oligonucleotides that exhibit a high degree of complementarity to a target nucleic acid sequence in a given genome and a lower degree of complementarity to other nucleic acid sequences. Those skilled in the art will recognize that a wide range of gene regions suitable for use in the present invention can be selected. An antisense compound is "specifically hybridizable" when the binding of the compound to the target nucleic acid interferes with the normal function of the target nucleic acid to modulate function and/or activity, and is sufficiently complementary to avoid antisense 4 conjugate Non-specific with non-dry nucleic acid sequences under conditions where specific binding is desired to occur (ie, under physiological conditions in the case of in vivo analysis or therapeutic treatment, and under conditions of analysis in the context of in vitro analysis) Combine. 156342.doc -83· 201143782 The hybridization properties of the oligonucleotides described herein can be determined by one or more in vitro assays known in the art. For example, the nature of the nucleotides described herein can be obtained by measuring the binding strength between the target natural antisense molecule and the potential drug molecule using a melting curve analysis. Any of the established methods of measuring the strength of the intermolecular interaction (e.g., melting curve analysis) can be used to estimate the binding strength between the crude natural antisense molecule and the potential drug molecule (molecule). Melting curve analysis measures the temperature at which a natural antisense molecule/molecular complex rapidly transitions from a double strand to a single stranded form. This temperature has been widely accepted as a reliable measure of the strength of the interaction between two molecules. Melt curve analysis can be performed using a cDNA copy of the actual native antisense RNA molecule or a synthetic DNA or RNA nucleotide corresponding to the molecular binding site. Multiple kits containing all necessary reagents to perform this assay (e.g., Applied Biosystems, MeltDoctor kit) can be used. Such kits comprise a suitable buffer solution containing one of a double stranded DNA (dsDNA) binding dye (e.g., ABI HRM dye, SYBR Green, SYTO, etc.). The nature of the dsDNA dye is that it emits almost no fluorescence in free form, but has high fluorescence when bound to dsDNA.

To perform the analysis, the cDNA or corresponding oligonucleotide is mixed with the molecule at a concentration defined by a particular manufacturer's protocol. The mixture was heated to 95 °C to dissociate all of the preformed dsDNA complexes and then slowly cooled to room temperature or other lower temperatures as defined by the kit manufacturer to anneal the DNA molecules. The newly formed composite was then slowly heated to 95 ° C while continuing to collect data on the amount of fluorescence produced in the reaction. The fluorescence intensity is inversely proportional to the amount of dsDNA present in the reaction 156342.doc •84- 201143782. Data can be collected using a real-time PCR instrument compatible with the kit (eg, ABI's StepOne Plus Real Time PCR System or lightTyper instrument, Roche Diagnostics, Lewes, UK). Draw a plot of the negative derivative of fluorescence relative to temperature (-d (fluorescence)/dT), y-axis) and temperature (X-axis) by using appropriate software (eg lightTyper (Roche) or SDS Dissociation Curve, ABI) To build a melting peak. The data was analyzed to determine the temperature at which the dsDNA complex rapidly converted to a single-stranded molecule. This temperature is called Tm and is proportional to the strength of the interaction between the two molecules. Typically, the Tm is above 40 °C. Example 2: Modulation of LHX2 Polynucleotide HepG2 cells were treated with antisense nucleoside acid. All antisense oligonucleotides used in Example 2 were designed as described in Example 1. The manufacturer (IDT Corporation, Coralville, IA) was instructed to manufacture the designed phosphorothioate-bonded oligonucleotides and to provide the designed phosphorothioate analogs shown in Table 1. The star symbol between the nucleotides indicates the presence of a phosphorothioate bond. The oligonucleotides required for the experiments in Example 2 can be synthesized using any suitable state of the art, such as the method used in IDT: the use of phosphorous ruthenium on solid supports (e.g., 5 micron fixed glass beads (CPG)) Amine monomer (common nucleotides, all of which are protected by protecting groups such as triphenylsulfonyl groups on sugars, benzoquinones on A and C, and N-2-isobutyl) on G) . The protecting group prevents undesired reactions from occurring during oligonucleotide synthesis. The protecting group is removed at the end of the synthesis process. The initial nucleotide is attached to the solid support via 3' carbon and synthesized in the 3' to 5' direction. Add a new base to the growing oligonucleotide strand in the following 4 steps: 1) Remove the protecting group from the 5' oxygen of the bitter acid using a tri-glycolic acid from the immobilized core 156342.doc -85- 201143782; 2) Use The tetranucleotide couples the immobilized nucleotide to the next nucleotide in the sequence; the reaction is continued via the tetrazolylphosphinamide intermediate; 3) the unreacted free nucleotide and the reaction are removed by washing By-product and capping the unreacted fixed oligonucleotide to prevent its participation in the next round of synthesis; by using acetic anhydride and N-methylimidazole, the free 5, hydroxyl group is acetylated to achieve capping; 4) Stabilize the bond between nucleotides, use Egyptian water (if you want to produce acid diester bonds), or use # reagent (3H1,2_ benzodithiol-3-keto-1,1-dioxide) (If a phosphorothioate bond is desired) to oxidize phosphorus. The chimeric backbone can be constructed by alternately using two oxidizing agents. The above 4-step cycle is repeated for each nucleotide in the sequence. Upon synthesis of the complete sequence, the acid is removed from the solid support using ammonium hydroxide at elevated temperature and deprotected. The protecting group is removed by washing with desalting and the remaining oligonucleotide is lyophilized. To carry out the experiment designed in Example 2, the growth medium (MEM/EBSS (Hyclone catalog number: SH30024, or Mediatech catalog number: MT-10-〇l〇-CV) + l at 37 ° C and 5% CO 2 〇% FBS (MediateCh catalog number: MT35-011-CV) + penicillin/streptomycin (Mediatech catalog number: Mt3〇-〇〇2-CI)) grew HepG2 cells from ATCC (catalog) No.: HB-8065). One day before the experiment, the cells were plated again in a 6-well plate at a density of 0.5 x 104 / ml and incubated overnight at 37 ° C and 5% C 〇 2 . On the day of the experiment, the medium in the 6-well plate was replaced with fresh growth medium. Oligonucleotides delivered by the manufacturer in the form of Donggan were diluted to a concentration of 2 〇 μΜ in deionized water without RNAse/DNAse. 2 μΐ 156342.doc -86 - 201143782 This solution was incubated with 400 μΐ OptiMEM medium (Gibco catalog number: 31985-070) and 4 μL Lipofectamine 2000 (Invitrogen catalog number: 11668019) for 20 min at room temperature, then drop by drop Apply to a well of a 6-well plate with HepG2 cells. A similar mixture containing 2 μM water instead of the nucleotide solution was used as a mock transfection control group. After incubation for 3-18 h at 37 ° C and 5% CO 2 , the medium was changed to a fresh growth medium. After 48 h of antisense nucleotide addition, the medium was removed and the SV Total RNA Isolation System from Promega (catalog number: Z3105) or the RNeasy Total RNA Isolation kit from Qiagen (catalog number: 74181) was used according to the manufacturer's instructions. RNA is extracted from cells. 600 ng of extracted RNA was added to the reverse transcription reaction using the Verso cDNA kit from Thermo Scientific (catalog number: AB1453B) or the high capacity cDNA reverse transcription kit (catalog number: 43688 13), as in the manufacturer's protocol. Said. The cDNA from this reverse transcription reaction was used by real-time PCR using ABI Taqman Gene Expression Mix (catalog number: 4369510) and primers/probes designed by ABI (Applied Biosystems Taqman Gene Expression Assay: Hs00180351_ml (Human) 5 Applied Biosystems, Foster City CA) to monitor gene expression. The following PCR cycle was performed using a StepOne Plus real-time PCR machine (Applied Biosystems): 2 min at 50 °C, 10 min at 95 °C, 40 cycles (15 seconds at 95 °C, at 60 °C) Hold at °C for 1 min). The fold change in gene expression after treatment with antisense nucleotides was calculated based on the difference between the 18S normalized dCt values between the treated and mock transfected samples. Sensitive shoulder: Real-time PCR results showed that the content of LHX2 mRNA in HepG2 cells was significantly increased after 48 h treatment with an oligomer designed to be LHX2 antisense 156342.doc -87-201143782 smaplerbu.aApr07 (Fig. 1). 373 cells were treated with antisense nucleotides at 37 ° C and 5 ° /. C02 under growth medium (MEM/EBSS (Hyclone catalog number: SH30024, or Mediatech catalog number: MT-10-010-CV) + 10% FBS (Mediatech catalog number: MT35-011_CV) + penicillin/streptomycin (Mediatech) Catalog number: MT30-002-CI)) 3T3 cells from ATCC (Catalog No.: CRL-1658) were grown. One day before the experiment, the cells were plated again in a 6-well plate at a density of 1.5 x 105/ml at 37 ° C and 5 ° /. C〇2 is cultivated. On the day of the experiment, the medium in the 6-well plate was changed to fresh growth medium. All antisense oligonucleotides were diluted to a concentration of 20 μΜ. 2 μL of this solution was incubated with 4 μμΐ 〇pti-MEM medium (Gibco Cat. No. 31985-070) and 4 μL Lipofectamine 2000 (Invitrogen Cat. No. 11668019) at room temperature for 20 min and applied to 6 wells. The plate has 3T3 cells in each well. A similar mixture containing 2 μM water instead of the oligonucleotide solution was used as a mock transfection control group. At 37. (: and after culturing for 3-18 h at 5% C02, the medium was changed to fresh growth medium. After adding antisense oligonucleotide for 48 h, the medium was removed and the SV Total RNA Isolation System from Promega was used. Z3 105) or RNeasy Total RNA Isolation kit from Qiagen (Catalog No. 741 81) Extract RNA from cells according to the manufacturer's instructions. Add 600 ng of RNA to the Verso cDNA kit from Thermo Scientific (Catalog No.: AB1453B) Or the reverse transcription reaction carried out by the high-capacity cDNA reverse transcription kit (catalog number: 43 688 13), as described in the manufacturer's section 156342.doc *· 88 · 201143782. The cDNA from this reverse transcription reaction is used. Real-time PCR uses ABI Taqman Gene Expression Mix (catalog number: 43695 10) and ABI designed primer/probe (Applied Biosystems Taqman Gene Expression Assay: Mm008 39783_ml (Mouse), Applied Biosystems, Foster City CA) to monitor gene expression The following PCR cycle was performed using a StepOne Plus real-time PCR machine (Applied Bio systems): kept at 50 ° C for 2 min, Hold at 95 ° C for 10 min, 40 cycles (15 seconds at 95 ° C, 1 min at 60 ° C)., Crust: Real-time PCR results show that the design is LHX2 antisense molecule After 48 h of oligomer treatment, the amount of LHX2 mRNA in 3T3 cells was significantly increased (Fig. 2). Although the invention has been illustrated and described in terms of one or more embodiments, those skilled in the art are reading and understanding Equivalent changes and modifications may be made in the specification and the drawings. Further, although specific features of the invention may be disclosed in accordance with one of the various embodiments, the features may be combined with one or more other features of other embodiments. This disclosure may be desirable and advantageous for any given or specific application. The Abstract of the Disclosure allows the reader to quickly ascertain the nature of the technical disclosure. This summary is based on the understanding that it is not intended to be construed Or limit the scope or meaning of the following patent application scope. [Simplified Schematic] Figure 1 shows the use of phosphorothioate oligosaccharide introduced using Lipofectamine 2000 compared to the control group. The fold change and standard deviation of 156342.doc-89-201143782 LHX2 mRNA in HepG2 cells at 48 hours after treatment. Real-time PCR results showed that one of the concentrators designed using LHX2 antisense smaplerbu.aApr07 was treated. At 48 h after the hour, the content of LHX2 mRNA in HepG2 cells increased significantly. The bars denoted as CUR-1560 to CUR-1563 correspond to the samples treated with SEQ ID NOS: 5 to 8, respectively. Figure 2 shows the fold change and standard deviation of LHX2 mRNA in 3T3 cells at 48 hours after treatment with the phosphorothioate oligomer introduced by Lipofectamine 2000 compared with the control group. Real-time PCR results show that the design is The content of LHX2 mRNA in 3T3 cells was significantly increased 48 h after treatment of one of the LHX2 mouse antisense molecules. The bars denoted as CUR-1588 to CUR-1595 correspond to the samples treated with SEQ ID NOS: 9 to 16, respectively. BRIEF DESCRIPTION OF THE SEQUENCE LISTING - SEQ ID NO: 1 : Homo sapiens LIM homolog 2 (LHX2), mRNA (NCBI accession number: NM_004789); SEQ ID NO: 2: native LHX2 antisense sequence (smaplerbu.aApr07); SEQ ID NO : 3 : mouse native LHX2 antisense sequence (fl〇ysla.aSep07); SEQ ID NO.. 4: mouse native LHX2 antisense sequence (CJ098619); SEQ ID NOs: 5 to 16: antisense oligonucleotide . * indicates thiophosphate vinegar bond. 156342.doc 90- 201143782 Sequence Listing <11〇> American Okokena Limited Company <120> Treatment of LHX2 by Inhibition of Natural Antisense Transcript Against LIM Homologous Sequence 2 (LHX2) Disease <130> LHX2 <140>100117637 <141>2011/05/19 <150> 61/346,284 <151>2010/05/19 <160> 16 <170> Patent In version 3.5 <;210> 1 <21!> 2416 <212> DNA <213> Homo sapiens

<300><3〇8> mjmm3 <309> 2011-03-12 <313> (1)..(2416) <400> 1 gactgcagag ccggggctgg gctaggcgcg cgcttggaga gcattgcgcg cggctgggcc 60 cgcggccggc ggctcctcct cccactctgc tcctcctctt ttttctcctc ctccacctcc 120 tcctccgcct cctcctcctc ctcttcctcc icctcttcaa ttctcccggt ggctcgactc 180 ggctcgcagg cttcggagaa acccctactc cagtcgccga ctcagcgccc aagagggtcg 240 ccttgggctg ggggcgcacc ccagggaggg gaggggtcca ggcagctggg ccgccgcgga 300 cacctagcgg cttcagggtg aaccccgacc gcagccgtcg ccgcctcggg cagagtttgc 360 gcccttgctt tgcgccccgg gcgctgaagc cgggcgggcg atgcccgcgg cgtgaaagcg 420 cccgcggcgg gcgccgacct ctgtcctagt ctcctgctcc ccccgccccg cttgtcccgt 480 gcccttgtga ccctggcttt ggcgccgtcg cccaggcgcc ccgcaatgta gctgcccctg 540 cgcctcggcg ggaggcgtcc tgccccgcga gcgcccgggg cccggagccc ggcctggggg 600 ctcagccgag ctcgggcggg gccggggccg cggtggcgat gcaccgggcc cgttagcgcc 660 aggagcgcca ggcagctgag gcggggggca agccctccct cggaggagcc gcgcccccgg 720 ccccgccggt cccgccgcga tgctgttcca cagtctgtcg ggccccgagg tgcacggggt 78 0 catcgacgag atggaccgca gggccaagag cgaggctccc sccatcagct ccgccatcga 840 ccgcggcgac accgagacga ccatgccgtc catcagcagt gaccgcgccg cgctgtgcgc 900 cggctgcggg ggcaagatct cggaccgota ctacctgctg gcggtggaca agcagtggca 960 catgcgctgc ctcaagtgct gcgagtgcaa gctcaacctg gagtcggagc tcacctgttt 1020 cagcaaggac ggtagcatct actgcaagga agactactac aggcgcttct ctgtgcagcg 1080 ctgcgcccgc tgccacctgg gcatctcggc ctcggagatg gtgatgcgcg ctcgggactt 1140 ggtttatcac ctcaactgct tcacgtgcac cacgtgtaac aagatgctga ccacgggcga 1200 ccacttcggc atgaaggaca gcctggtcta ctgccgcttg cacttcgagg cgctgctgca 1260 gggcgagtac cccgcacact tcaaccatgc cgacgtggca gcggcggccg ctgcagccgc 1320 ggcggccaag agcgcggggc tgggcgcagc aggggccaac cctctgggtc ttccctacta 1380 caatggcgtg ggcactgtgc agaaggggcg gccgaggaaa cgtaagagcc cgggccccgg 1440 156342- sequence Listing .doc 201143782 tgcggatctg gcggcctaca acgctgcgct aagctgcaac gaaaacgacg cagagcacct 1500 ggaccgtgac cagccatacc cgagcagcca gaagaccaag cgcatgcgca cgtccttcaa 1560 gcaccaccag Cttcggacca tgaagtctta ctttgccatt aac cacaacc ccgacgccaa 1620 ggacttgaag cagctcgcgc aaaagacggg cctcaccaag cgggtcctcc aggtctggtt 1680 ccagaacgcc cgagccaagt tcaggcgcaa cctcttacgg caggaaaaca cgggcgtgga 1740 caagtcgaca gacgcggcgc tgcagacagg gacgccatcg ggcccggcct cggagctctc 1800 caacgcctcg ctcagcccct ccagcacgcc caccaccctg acagacttga ctagccccac 1860 cctgccaact gtgacgtccg tcttaacttc tgtgcctggc aacctggagg gccatgagcc 1920 tcacagcccc tcacaaacga ctcttaccaa ccttttctaa tgactcgcaa cccctcaccc 1980 cacaatttct ttaaaaaaga aattatcttt agttgaattc caagtgtalt ttaaaataga 2040 ggctttgagc aactaaciaa ccacatttta ggztctcgcc tggaaacaga ggtaaaaaaa 2100 agaagtgtgc gcccggctaa tgcagcggtg t £ gaccgag £ aacaacitgg aagatctacc 2160 tgcaacacaa catttgtgtc actgtacagt ittgtggact gagcgaggaa aaacaacaaa 2220 taatttaagt tggctagagc ttctgtattt tcaaagactg ccacgtgcct taggaatact 2280 gttttatctc catactttgg atgacttgtt catttttctc tccctctttt tctctgtata 2340 ttiatgacca gagcaaaaat gtaaaaaaca aaaaaaacaa caaaaaaagt ttgttacttt 2400 gaatagtcct aaaaag 2416 <210> 2 <211> 21 45 < 212 > DMA < ^ 13 > Homo sapiens < 400 > 2 tacatatagg cacgcgattc cgtcgccacc tctcggacca gcagcccacg tccaacgctg 60 ggccgacccc agatacacag gcagtcggga ttcccgcccg gtgcgcttgt ctattcatcc 120 ctctgcgtca ggctgggacg cgctccgtct gtaaaaggct caaacgcatc tcccgccgcg 180 gggcgggatc taggggccca ggccccgggg tccagaggcg ggtaactttg ctaatctccc 240 ccagcggcgg ggaacgtcgc gcaaccgctg agccctgtcc gccgagacta aacaagcaga 300 ggaacaggct gtaaacacac atcctgacac gcaggsatgt tgctcttgga gaatgtgaag 360 acagtttgct tcttgacaag cgagcgaacg ggcgcccaga tttttgcagc ctttccgagt 420 ctcccccgag ggagaggcgg cagagaaaac gccggatttg ggagccacca gggaaggatc 480 cgcgcagggg agccgccctc cttggcccca gacccgcctg cctggggccc cctttgctca 540 ctgtcaatgt atggtctgaa gctctgagga tggtgctggg gactggggtc gggggaagcc 600 tcttgaataa taactgcaaa gaagaaagag gcgagaacgt ctccctaacc ttgaagcaga 660 aaggactgtg ttcttaaagc tgttggctgc agtcacaggg ccagttgccc Gcctctgttc 720 cctgagtaaa gtgtaacatc ttctgtcctc ctggctt|:ct tgcaccattc a2caaattat 780 actccttcct taccaaagtg ggaatgctca ggga agtgtg igtgtgtgtg tgtgtgtgtg 840 tgtgtcccct ctgcactgag gctgctgtta gagatgttac caatttaaac cttccagaat 900 cctggaggtt tacattttgt aaagggaggg gacactcctg gactgtcaca atcccaattc 960 tggttagagt ggcagggact gaacaaagcc accaacatgc aaaagtccat ctccagagta 1020 -2 · 156342- Sequence Listing .doc 201143782 ❹ gcatgcatgc taacctctgt tgcagggtta aaaagctctg tcattcagca tcagttcagg cacagaaaat ccgaagagga cagagagaag cgcctccctg cctgttaaat atgcatcagg gtattaatat atgcaatttt gagggaaatg caaagtggtg iaggtcagag gtatgtatct ttttctgaaa ccccaggaaa ctcctgtttg gggttatctc tatcttcaat aagcagggag caugctatc ctgcttatca accccgtgtc cccaattcct ggcgtcagct gcactgaact aaacgtggag tstattaaat gtcaaggttt aggggctgct acaaaatglt attgttcacc aggtcctgct gaacaaitat cccccagtgg taaggcagag caagaaaggg gaggacccac tgttttatat tctttcttta ccgctatcga tgtgacatta cctcctcttc gagcaggttg taaaatgaaa gttggcagcc gttatgagaa ttatctgtga ggcatgtgac caattttatt tcagcaaaag taaatcacat aacaaaatac gatatgcttc gaagggtgat tggggtgagg acacacacac tgatgcgasa cgaaatcagc caataaaaat Gaatagataa ic Cctgcagc gggaactaac cacgaagtgt cttttcctcc agtaaaggct ttatgattcc atgtgtttta acagggaatt gatttaccat tgtcaacaat actatagcta ctgcactttc tccttgccac ctgagtcaca ctttcccagg ggctgtcagt tcattgcctt

Ugtggagcc gtggcttggc gatctsaaca cagggctctc gaatttc ^ gg ctcctgcttt gcggagagga agatgtagaa aggtgtttgg SStaaaagaa tattgattag ataaatctgt attgc cccttctctc tgcacaggaa gggagagcag cttgtgggcc cagcagtaaa ggtcacacta ttagttgtti ctgttgcagg attctgaaac tctctagggc tttgaacatg tcagtagcag aigtgctcag actcccggag cagtgtttct atatgaatac ggtgcagaaa cagatcagat 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 I860 1920 1980 2040 2100 2145 Ο < 210 > 3 < 211 > 606 < 212 > DNA < 213 > Mus musculus < 400 > 3 actggaccat gagtgttgtg gatgaaagct ttgctaigta tgggtccaag tgacaatccc taactcctgg cttcattact tataaaataa ggggggggag ttaacatacg tgcatggatc gcatccgccg agcacagagt caglggcacg gctggatatg gcagtggatt gaaccttgga taafttcaag agcagcagtg Tcctta gctgtaatgt ttgccggaac gcgctttgaa aactgtaaac gatgttgaaa caatcacatc atcaagttgt gagaaaatga aaacaaaacg gagtcttttt gctggtggtg cctcctgctc agatccctgg caaagctcac iccctaccaa atttcatttt cacccacatc agctcactga aaacacccag ggatgccaga acaaatggaa tgtcttctgc actcaagaac tgaacc Tcag agacacaatg ctccttttga acctcagtag aaatgggacc aagccatcat ggctgcacta tmtgacgt attctgcagc agtctgggct tttccccaac ctatgtgaca tatccagtaa cacctcccat tgtctaggaa gtcgcaagaa aatglctttc 60 120 180 240 300 360 420 480 540 600 606 0>1>2>3> 4 480 DNA Mus musculus <220> 156342 - Sequence Listing .d〇c 201143782 <221> misc feature <222> (355)..(355) <223> n is a, c, g, or t <400> 4 gtgcggagat gccgagcgca aacttcccgg ggctcccggg gcaggcgggg ggggccgggg agggaggaag cgcccagccc aggsagaaag cagggatgtt tatgtgaaga cagtttgctt tttgacaagc gagcaagcgg gcgcccagat tttggagtct ccccggaggg aaaggcagat agaacaaacc tcggactggg ggaagggctc tgcgcagggg aaccgcccca ccctttggcc ccagactctc ctccggtsgt actactaaag gattgcctca agctctgagg atgaagctgg tccggaaacc tctcgaataa caactgtggg ggagaaagag actctgccaa tggcctcaca acggaaagga ctgttttctt aaagctgttg acttcagtca < 210 > 5 < 211 > 19 < 212 > leg < 213 > artificial sequence <220><223> antisense oligonucleotide <400> 5 gtccctgcca ctctaacca <210≫ 6 <211> 20 <212> m < 213 > artificial sequence <220><223> antisense nucleotide <400> 6 ttctgctctc cctgtgactc

<210> Ί <211> 20 <212> DNA <213> artificial sequence <220><223> antisense nucleotide <400> 7 tttctctgcc gcctctccct <210> 8 <211> 20 <212> DNA <213>Artificial sequence<220><223> Antisense nucleotides <400> 8 cttccctgag cattcccact <210> 9 <211> 21 <212> m artificial Sequence-4 - ccacccggcc actcttggag ttctacagcc gagccatcct ctgcctgggg ggttnagggt catttcttcc cagagccagt 156342 - Sequence Listing.doc 201143782 <220><223> Antisense Oligonucleotide <400> 9 gttctatctg cctttccctc c <210> 10 <211> 20 <212> ΪΜΚ <213> artificial sequence <220><223> antisense oligonucleotide <400> 10 acatccctgc tttctccctg <210> 11 <211> 21 <212>

<213>Artificial sequence <220>

<223> Antisense Oligonucleotide <400> II tttagtagta ccaccggagc c <210><211><212><213> 12 20

DNA artificial sequence <220> antisense oligonucleotide <400> 12 cccagcttca tcctcagagc <210> 13 <211> 21 <212>leg <213> artificial sequence <220><^3> Antisense nucleotides <400> 13 ctttccgttg tgaggccagg a <210> 14 <2Π> 21 <212> Wk <213> artificial sequence <220><223> antisense oligonucleotide <223>;400> 14 tccatatcca gccgtgccac t <210> 15 <211> 21 <212> Leg <2]3>Artificial sequence <220><223> Antisense oligonucleotide 156342 - Sequence Listing.doc 201143782 <400> 15 tccatttgtc accaccagca c 21 <210> 16 <211> 20 <212> Wk <213>Artificial sequence<220><223> Antisense oligonucleotide <400> Ggcttctggg tgtttcgttt 20 156342 - Sequence Listing.doc

Claims (1)

201143782 VII. Patent Application Range: 1. Use of an antisense oligonucleotide of 5 to 30 nucleotides in length for the production of LIM homologous sequence 2 (LHX2) polynucleotide in a biological system An agent that functions and/or exhibits acidity wherein the nucleotide has at least 50% sequence identity to the reverse complement of the natural antisense transcript of the LIM homolog 2 (LHX2) polynucleotide. 2. The use of claim 1, wherein the oligonucleotide comprises nucleotides 1 to 2145 comprising SEQ ID NO: 2 of the natural antisense transcript, nucleotides 1 to 606 of SEQ ID NO: 3, and SEQ ID NO: The reverse complement of the 5 to 30 contiguous nucleotides of nucleotides 1 to 480 of 4 has at least 5 Å. Sequence Consistency of 〇/ 3. 3. The use of an antisense nucleotide of 5 to 30 nucleotides in length to modulate LIM homologous sequence 2 (LHX2) aggregation in a patient's cells or tissues An agent that functions and/or exhibits a nucleotide, wherein the oligonucleotide has at least 50% sequence identity to an antisense oligonucleotide of the LIM homolog 2 (LHX2) polynucleotide. 4. The use of claim 3, wherein the nucleotide and nucleotides 1 to 2145 comprising SEQ ID NO: 2 of the natural antisense transcript, nucleotides 1 to 606 of SEQ ID NO: 3, and SEQ ID NO The reverse complement of a polynucleotide of 5 to 30 contiguous nucleotides within nucleotides 1 to 480 of 4 has a sequence identity of at least 50°/〇. 5. Use of an antisense oligonucleotide targeting a region of a natural antisense oligonucleotide of a LIM homolog 2 (LHX2) polynucleotide for use in the regulation of LIM in a biological system Function of source sequence 2 (LHX2) polynucleotide 156342.doc 201143782 and/or agent of expression. 6. The use of claim 5, wherein the function and/or performance of the LIM homolog 2 (LHX2) is enhanced compared to the control group. 7. The use of claim 5, wherein the antisense raised nucleotide targets a natural antisense sequence of LIM homolog 2 (LHX2) polynucleotide. 8. The use of claim 5, wherein the antisense oligonucleotide targets a nucleic acid sequence comprising a coding and/or non-coding nucleic acid sequence of a LIM homolog 2 (LHX2) polynucleotide. 9. The use of claim 5, wherein the antisense raised nucleotide targets an overlapping and/or non-overlapping sequence of LIM homologous sequence 2 (LHX2) polynucleotide. 10. The use of claim 5, wherein the antisense oligonucleotide comprises one or more modifications selected from the group consisting of at least one modified sugar moiety, at least one modified internucleoside linkage, at least one modified Nucleotides, and combinations thereof. 11. The use of claim 10, wherein the one or more modifications comprise at least one modified sugar moiety selected from the group consisting of 2, _o-methoxyethyl modified sugar moieties, 2'-methoxy Modified sugar moiety, sugar moiety modified with 2,_〇-alkyl, bicyclic sugar moiety, and combinations thereof. 12. The use of claim 10, wherein the one or more modifications comprise at least one modified internucleoside linkage selected from the group consisting of: phosphorothioate, 2,-methoxyethyl (MOE), 2 '-Fluorine, alkyl phosphonate, dithiophosphate, thioester thiodecanoate, amino phosphate, urethane, acid ester, glycerate, amino acetate, Carboxynonyl esters, and combinations thereof. 13_ The use of claim 10, wherein the one or more modifications comprise at least one modified nucleotide selected from the group consisting of: peptide nucleic acid (pNA), locked nucleic acid 156342.doc 201143782 (LNA), arabinic acid (FANA) ), analogs, derivatives, and combinations thereof. 14. As requested! Use of the oligonucleotide, wherein the oligonucleotide comprises at least one oligonucleotide sequence as shown in seq ID NO: 5 to 16. 15. Use of a short interfering 11]^8 (siRNA) nucleotide of 5 to 3 nucleotides in length for the manufacture of a medicament for modulating LIM homology in mammalian cells or tissues The function and/or expression of the sequence 2 (LHX2) gene, wherein the siRNA oligonucleotide is specific for an antisense polynucleotide of a LIM homolog 2 (LHX2) polynucleotide, and wherein the at least one siRNA The raised nucleotide has at least 50% sequence identity to the antisense of the LIM homolog 2 (LHX2) polynucleotide and/or the complement of at least about 5 contiguous nucleic acids of the sense nucleic acid molecule. 16. The use of claim 15, wherein the oligonucleotide is at least about 5 contiguous nucleic acids of the antisense of the um homologous sequence 2 (LHX2) polynucleotide and/or the complement of the sense nucleic acid molecule. The sequence has a sequence identity of at least 80%. 17. Use of an antisense oligonucleotide of about 5 to 3 nucleotides in length, which is sensed in a LIM homolog 2 (LHX2) polynucleotide and And/or a non-coding and/or coding sequence of a natural antisense strand, which is used to produce a function for modulating LIM homolog 2 (LHX2) in a mammalian cell or tissue and/or An agent for expression, wherein 4 at least one antisense oligonucleotide has at least 5% sequence identity with at least one nucleic acid sequence as set forth in SEq ID N〇: 1 to 4. 18. An oligonucleotide comprising at least one modified synthesis and modification, wherein at least one of the 156342.doc 201143782 is selected from the group consisting of a modified sugar moiety, at least one modified core An internucleotide linkage, a modified nucleotide, and a σ thereof, wherein the oligonucleotide is hybridized with the LIM homolog 2 (Lhx2) gene in the tongue or in vitro and The antisense compound of the read homologous sequence 2 (LHX2) gene and/or the expressed antisense compound can be modulated relative to the normal control group and wherein the nucleotide and the UM homologous sequence 2 (lhx2) polynucleotide are A sequence of at least about 5 contiguous nucleic acids of an antisense and/or sense nucleic acid molecule and its alleles, homologs, isoforms, variants, derivatives, mutants, fragments, or combinations of complementary sequences having at least 5 〇% sequence consistency. 19. The nucleotide of claim 18, wherein the oligonucleotide is 5 to 30 nucleotides in length and 5 to 30 consecutive nucleotides in the natural antisense transcript of the LHX2 gene The reverse complement sequence has a sequence identity of at least 5%. 20. The oligonucleotide of claim 19, wherein the at least one modification comprises an internucleotide linkage selected from the group consisting of phosphorothioate, alkyl phosphonate, dithiophosphate, sulfur An alkyl phosphonate, an amino phosphate, a urethane, a carbonate, a phosphate, an amino acetate, a carboxy thioglycol, and combinations thereof. 21. The nucleotide of claim 19, wherein the nucleotide comprises at least one phosphorothioate internucleotide linkage. 22. The oligonucleotide of claim 19, wherein the nucleotide comprises a backbone of a phosphorothioate internucleotide linkage. 23. The oligonucleotide of claim 19, wherein the raised nucleotide comprises at least one 156342.doc 201143782 modified nucleotide selected from the group consisting of: a peptide nucleic acid, a locked nucleic acid (LNA), Analogs, derivatives, and combinations. 24. The nucleotide of claim 19, wherein the nucleotide comprises a plurality of modifications wherein the modifications comprise modified nucleotides selected from the group consisting of phosphorothioates, alkyl phosphonates, A phosphorothioate, an alkyl thiophosphonate, an amino phosphate, an amino phthalate, a carbonate, a phosphate, an aminoacetic acid, a buffer, and combinations thereof. 25. The oligonucleotide of claim 19, wherein the nucleotide comprises a plurality of modifications wherein the modifications comprise modified nucleotides selected from the group consisting of peptide nucleic acids, locked nucleic acids (LNA), analogs thereof , derivatives, and combinations. 26. The nucleotide of claim 19, wherein the oligonucleotide comprises at least one modified sugar moiety selected from the group consisting of a 2,_〇-methoxyethyl modified sugar moiety, via 2'- a methoxy-modified sugar moiety, a 2,_〇-alkyl-modified sugar moiety, a bicyclic sugar moiety, and combinations thereof. 27. The oligonucleotide of claim 19, wherein the raised nucleotide comprises a plurality of modifications wherein the modifications comprise a modified sugar moiety selected from the group consisting of 2'-〇-methoxyethyl modified a sugar moiety, a 2,0-oxy-modified sugar moiety, a 2'-0-alkyl modified sugar moiety, a bicyclic sugar moiety, and combinations thereof. 28. The oligonucleotide of claim 19, wherein The oligonucleotide is at least about 5 to 30 nucleotides in length and hybridizes to an antisense and/or sense strand of a LIM homolog 2 (LHX2) polynucleotide, wherein the oligonucleotide is associated with the LIM An antisense and/or sense encoding in a homologous sequence 2 (LHX2) polynucleotide and/or a complementary sequence of at least about 5 contiguous nucleic acids in a non-coding nucleic acid sequence having at least about 156342.doc 201143782 60% sequence identity Sex. 29. The nucleotide of claim 19, wherein the nucleotide is at least in the antisense and/or sense encoding and/or non-coding nucleic acid sequence of the lim homologous sequence 2 (LHX2) polynucleotide A complementary sequence of about 5 contiguous nucleic acids has a sequence identity of at least about 80%. 30. The nucleotide of claim 19, wherein the nucleotide is hybridized in vivo or in vitro to at least one LIM homolog 2 (LHX2) polynucleotide and can be modulated relative to a normal control group. The performance and/or function of nucleotides. 31. The nucleotide of claim 19, wherein the oligonucleotide comprises a sequence as set forth in ID NO: 5 to 16. 32. A pharmaceutical composition comprising: or a plurality of oligonucleotides specific for one or more LIM homolog 2 (LHX2) polynucleotides of claim 18 and a pharmaceutically acceptable form Agent. 33. The composition of claim 32, wherein the oligonucleotides are aligned with any one of the nucleotide sequences, such as seq N〇: 5 to 16, having at least about 40°. Sequence consistency. 34. The composition of claim 32, wherein the oligonucleotides comprise nucleotide sequences such as seq NO 5 to 16. 35. The composition of claim 34, wherein the I nucleotides as indicated by seq id no: 5 to 16 comprise one or more modifications or substitutions. 36. The composition of claim 35, wherein the one or more modifications are selected from the group consisting of thiophosphoric acid, phosphonium phosphonate, peptide nucleic acid, locked nucleic acid (LNA) molecules, and combinations thereof. 156342.doc 201143782 37. 38.t) 39. Ο 40. Use of an antisense oligonucleotide for the manufacture of a polynucleotide for preventing or treating at least one LIM homologous sequence 2 (LHX2) and Or an agent thereof, or at least one agent encoding a disease, wherein the antisense oligonucleotide binds to a natural antisense sequence of the at least one UM-like homologous sequence 2 (LHX2) polynucleotide and modulates the at least one UM Expression of homologous sequence 2 (LHX2) polynucleotide. The use of claim 37, wherein the disease associated with the at least one LIM homolog 2 (LHX2) polynucleotide is selected from the group consisting of: a disease or condition associated with abnormal function and/or performance of [11 and 2, a blood disease Or a condition, a hair growth buckle, a neurological disease or condition, a retinal disease or condition, a disease or condition associated with impaired immunity, a pituitary disease or condition, a liver disease or condition, an olfactory system disease or condition, impaired erythropoiesis, Hematopoietic diseases or conditions, myeloproliferative diseases, and long limbs of the spine. A method for identifying and selecting at least one nucleotide that is selective for a natural antisense transcript of the LHX2 gene as a selected target polynucleotide for in vivo administration, the method comprising: identifying at least one of the inclusions and the selection An oligonucleotide of at least 5 contiguous nucleotides at least partially complementary to an antisense polynucleotide of a target polynucleotide; measuring the antisense nucleotide and the target polynucleotide under stringent hybridization conditions or The thermal melting point of the hybrid of the antisense polynucleotide of the selected target polynucleotide; and selecting at least one nucleoside acid for in vivo administration based on the information obtained. A method of modulating the function and/or expression of a UM homologous sequence 2 (LHX2) polynucleotide in a biological system in vitro comprising: rendering the system antisense with at least one 5 to 30 nucleotides in length Oligonucleotide contact, wherein the at least one oligonucleotide of 156342.doc 201143782 has at least 5% sequence identity with the reverse complement of the natural antisense of LIM homolog 2 (LHX2) polynucleotide, - thereby modulating the function and/or expression of the LIM homolog 2 (LHX2) polynucleotide. 41. The method of claim 40, wherein the biological system is in vitro modulating the function and/or expression of a LIM homolog 2 (LHX2) polynucleotide, comprising: causing the biological system to have at least one length of 5 to 3 Nucleotide antisense oligonucleotide contacts, wherein the at least one oligonucleotide is nucleoprotein 1 to 2145, SEQ ID NO: 3 comprising the natural antisense transcript of SEQ ID NO: 2 The reverse complement of the polynucleotide of 1 to 606 and 5 to 30 contiguous nucleotides within nucleotides 1 to 480 of SEQ ID NO: 4 has a sequence identity of at least 5〇0/〇; The function and/or expression of the LIM homolog 2 (LHX2) polynucleotide is modulated. 42. A method of modulating the function and/or expression of a LIM homolog 2 (LHX2) polynucleotide in a patient cell or tissue in vitro, comprising: cultivating a cell or tissue such as shai with at least one length of 5 Contacting an antisense oligonucleotide of up to 3 nucleotides, wherein the oligonucleotide has at least 5% sequence with the antisense oligonucleotide of the LIM homolog 2 (LHX2) polynucleotide Consistency ' thereby modulating the function and/or expression of the LIM homolog 2 (LHX2) polynucleotide in a patient's cells or tissues in vivo or in vitro. 43. A method of modulating the function and/or expression of a LIM homolog 2 (LHX2) polynucleotide in a patient cell or tissue, comprising: causing a biological system with at least one length of 5 to Contacting 3 nucleotides of antisense nucleotides, wherein the at least one nucleotide comprises nucleotides 1 to 2145, SEQ ID NO of SEQ ID NO: 2, including natural antisense 156342.doc 201143782 transcript The reverse complement of the nucleotides of nucleotides 1 to 606 of 3 and 5 to 30 contiguous nucleotides of nucleotides 1 to 480 of SEQ ID NO: 4 has at least 50% sequence identity Thereby modulating the function and/or performance of the LIM homolog 2 (LHX2) polynucleotide. 44. A method of modulating the function and/or expression of a LI]y [homologous sequence 2 (LHX2) polynucleotide in a biological system, comprising: targeting the system to at least one of the LIM homologous sequences 2 (LHX2) The antisense oligonucleotide of the region of the natural antisense nucleotide of the polynucleotide is contacted; thereby modulating the function and/or expression of the LIM homolog 2 (LHX2) polynucleotide. 45. The method of claim 44, wherein the LIM homolog 2 (LHX2) is enhanced in function and/or performance in vitro compared to a control group. 46. The method of claim 44, wherein the at least one antisense nucleotide targets a natural antisense sequence of a LIM homolog 2 (LHX2) polynucleotide. 47. The method of claim 44, wherein the at least one antisense oligonucleotide targets a nucleic acid sequence comprising a coding and/or non-coding nucleic acid sequence of a LIM homolog 2 (LHX2) polynucleotide. 48. A method of claim, wherein the at least one antisense nucleotide targets an overlapping and/or non-overlapping sequence of a LIM homolog 2 (LHX2) polynucleotide. The method, wherein the at least one antisense nucleoside acid comprises one or more modifications selected from the group consisting of: at least one modified sugar moiety, at least one modified core (four) linkage, at least one modified core (four) combination thereof . The method of claim 49, wherein the one or more (four) comprises at least one sugar selected from the group consisting of the following: saccharin, yoghurt, turmeric, 2-0-decyloxyethyl modified sugar 〇P knife, & 2 曱oxy modified sugar moiety, 2,_〇-alkyl modified sugar moiety 'bicyclic sugar moiety, and combinations thereof. 51. The method of claim 49, wherein the or more modifications comprise at least one modified internucleoside linkage selected from the group consisting of phosphorothioate '2'-0 methoxyethyl (MOE) , 2'-fluoro, alkyl phosphonate, dithiophosphate, thiophosphonic acid alkyl vinegar, amino citrate, amino citrate, carbonate, phosphate triester, amino acetate 52. The method of claim 49, wherein the one or more modifications comprise at least one modified nucleotide acid selected from the group consisting of: peptide nucleic acid (pNA), locked nucleic acid (LNA) And a method of claim 40, wherein the at least one raised nucleotide comprises at least one of the oligos shown in SEQ ID NOS: 5 to I6. Nucleotide sequence 54. A method of modulating the function and/or expression of a homologous sequence 2 (LHX2) gene in a mammalian cell or tissue in vitro, comprising: equating the cell or tissue with at least one length 5 to 30 nucleotides of short acid interfering RNA (siRNA) nucleoside acid exposure' of at least one siRNA oligonucleotide to LIM homologous sequence 2 (LHX2) The antisense polynucleotide of the polynucleotide is specific in that at least one siRNA oligonucleotide and at least one of the antisense and/or sense nucleic acid molecules in the lim homologous sequence 2 (LHX2) polynucleotide a complement of about 5 contiguous nucleic acids having a sequence identity of at least 5%; and modulating the function of the LIM homologue 156342.doc -10- 201143782 2 (LHX2) in a mammalian cell or tissue in vitro and/or 55. The method of claim 54, wherein the raised nucleotide is at least about 5 consecutive to the antisense of the LIM homolog 2 (LHX2) polynucleotide and/or the complement of the sense nucleic acid molecule. The sequence of the nucleic acid has a sequence identity of at least 80%. 56. A method of modulating the function and/or expression of UM homologous sequence 2 (LHX2) in a mammalian cell or tissue in vitro, comprising: constituting the cells Or the tissue is contacted with at least one antisense oligonucleotide having a length of about 5 to 30 nucleotides. 'The antisense oligonucleotide is sensed for LIM homolog 2 (LHX2) polynucleic acid and/or a non-coding and/or coding sequence of a natural antisense strand having specificity, wherein the at least one antisense oligonucleotide And the at least one nucleic acid sequence of SEQ ID NOS: 1 to 4 has a sequence identity of at least 5%; and modulating the function of the LIM homolog 2 (LHX2) in a mammalian cell or tissue in vitro And / or performance. 156342.doc