WO1999047655A2 - Sequences d'acides nucleiques humains provenant de tissus mammaires sains - Google Patents

Sequences d'acides nucleiques humains provenant de tissus mammaires sains Download PDF

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Publication number
WO1999047655A2
WO1999047655A2 PCT/DE1999/000909 DE9900909W WO9947655A2 WO 1999047655 A2 WO1999047655 A2 WO 1999047655A2 DE 9900909 W DE9900909 W DE 9900909W WO 9947655 A2 WO9947655 A2 WO 9947655A2
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WO
WIPO (PCT)
Prior art keywords
undef
seq
prostate
nucleic acid
breast
Prior art date
Application number
PCT/DE1999/000909
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German (de)
English (en)
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WO1999047655A3 (fr
Inventor
Thomas Specht
Bernd Hinzmann
Armin Schmitt
Christian Pilarsky
Edgar Dahl
André ROSENTHAL
Original Assignee
Metagen Gesellschaft Für Genomforschung Mbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Metagen Gesellschaft Für Genomforschung Mbh filed Critical Metagen Gesellschaft Für Genomforschung Mbh
Priority to JP2000536838A priority Critical patent/JP2002506639A/ja
Priority to EP99924684A priority patent/EP1070125A2/fr
Publication of WO1999047655A2 publication Critical patent/WO1999047655A2/fr
Publication of WO1999047655A3 publication Critical patent/WO1999047655A3/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

  • the invention relates to human nucleic acid sequences from breast tissue which code for gene products or parts thereof, their functional genes which code for at least one biologically active polypeptide and their use.
  • the invention further relates to the polypeptides obtainable via the sequences and their use.
  • Breast cancer is one of the main causes of death in women, and new therapies are needed to combat it. Therapies used so far, such as Chemotherapy, hormone therapy or surgical removal of the tumor tissue often do not lead to complete healing.
  • the phenomenon of cancer is often associated with the over- or under-expression of certain genes in the degenerate cells, although it is still unclear whether these altered expression rates are the cause or the consequence of the malignant transformation. The identification of such genes would be an essential step in the development of new therapies for cancer.
  • ESTs Expressed Sequence Tags
  • cDNAs ie reverse-transcribed mRNAs
  • the EST sequences are determined for normal and degenerate tissues.
  • Various operators offer such databases commercially.
  • the ESTs in the LifeSeq database used here are typically between 150 and 350 nucleotides long. They represent an unmistakable pattern for a particular gene, although this gene is usually much longer (> 2000 nucleotides).
  • the nucleic acid sequences Seq. ID No 1 to Seq. ID No.76 and Seq. ID No.161 to Seq. ID 178 can be found, which play a role as candidate genes in breast cancer.
  • the nucleic acid sequences Seq are of particular interest. ID Nos. 1-5, 10- 12, 14, 15, 19, 21-25, 28, 30, 31, 34, 37, 43, 45, 48, 50, 51, 58-65, 68, 69, 71 , 72, 74, 76 and 161-178.
  • the invention thus relates to nucleic acid sequences which encode a gene product or a part thereof, comprising a) a nucleic acid sequence selected from the group of
  • the invention further relates to a nucleic acid sequence according to one of the sequences Seq. ID Nos. 1-5, 10-12, 14, 15, 19, 21-25, 28, 30, 31, 34, 37, 43, 45, 48, 50, 51, 58-65, 68, 69, 71 , 72, 74, 76 and 161-178 or a complementary or allelic variant thereof and the nucleic acid sequences thereof which have a 90% to 95% homology to a human nucleic acid sequence.
  • the invention also relates to the nucleic acid sequences Seq. ID No. 1 to Seq. ID No. 76 and Seq. ID No.161 to Seq. ID 178, which are expressed in increased amounts in normal breast tissue or are expressed in reduced amounts in breast tumor tissue.
  • the invention further relates to nucleic acid sequences comprising a part of the above-mentioned nucleic acid sequences, in such a sufficient size that they can be combined with the sequences Seq. ID Nos. 1-5, 10-12, 14, 15, 19, 21-25, 28, 30, 31, 34, 37, 43, 45, 48, 50, 51, 58-65, 68, 69, 71 , 72, 74, 76 and 161-178 hybridize.
  • the nucleic acid sequences according to the invention generally have a length of at least 50 to 4500 bp, preferably a length of at least 150 to 4000 bp, in particular a length of 450 to 3500 bp.
  • expression cassettes can also be constructed in accordance with current process practice, with at least one of the nucleic acid sequences according to the invention together with at least one control or regulatory sequence known to the person skilled in the art, such as. B. a suitable promoter is combined.
  • the sequences according to the invention can be inserted in sense or antisense orientation.
  • Expression cassettes or vectors are to be understood: 1. bacterial, such as. B., phagescript, pBs, ⁇ X174, pBluescript SK, pBs KS, pNH8a, pNH16a, pNH18a, pNH46a (Stratagene), pTrc99A, pKK223-3, pKK233-3, pDR540, pRIT5 (Pharmacia), like eukaryont, 2nd eukaryont.
  • Suitable control or regulatory sequence means suitable promoters.
  • Two preferred vectors are the pKK232-8 and the PCM7 vector.
  • the following promoters are specifically meant: lad, lacZ, T3, T7, gpt, lambda PR, trc, CMV, HSV thymidine kinase, SV40, LTRs from retrovirus and mouse
  • the DNA sequences on the expression cassette can encode a fusion protein which comprises a known protein and a biologically active polypeptide fragment.
  • the expression cassettes are also the subject of the present invention.
  • the nucleic acid fragments according to the invention can be used to produce full-length genes.
  • the available genes are also the subject of the present invention.
  • the invention also relates to the use of the nucleic acid sequences according to the invention and the gene fragments obtainable from the use.
  • nucleic acid sequences according to the invention can be brought into host cells with suitable vectors, in which the heterologous part contains the genetic information contained on the nucleic acid fragments which is expressed.
  • the host cells containing the nucleic acid fragments are also the subject of the present invention.
  • Suitable host cells are e.g. B. prokaryotic cell systems such as E. coli or eukaryotic cell systems such as animal or human cells or yeasts.
  • the nucleic acid sequences according to the invention can be used in sense or antisense form.
  • the polypeptides or their fragments are produced by cultivating the host cells in accordance with common cultivation methods and then isolating and purifying the peptides or fragments, likewise by means of conventional methods.
  • the invention further relates to nucleic acid sequences which encode at least a partial sequence of a biologically active polypeptide.
  • the present invention relates to partial polypeptide sequences, so-called ORF (open reading frame) peptides, according to the sequence protocols Seq. ID Nos 77-85, 87, 88, 90, 91, 93, 95-108, 112-117, 119, 122, 124-126, 132, 133, 135, 137-160 and 179-209.
  • ORF open reading frame
  • the invention further relates to the polypeptide sequences which have at least 80% homology, in particular 90% homology, to the polypeptide partial sequences according to the invention of Seq. ID Nos 77-85, 87, 88, 90, 91, 93, 95-108, 112-117, 119, 122, 124-126, 132, 133, 135, 137-160 and 179-209.
  • the invention also relates to antibodies which are directed against a polypeptide or fragment thereof, which of the nucleic acids of the sequences Seq. ID No. 1 to Seq. ID 76 and Seq. ID No. 161 to Seq. ID 178 can be encoded.
  • Antibodies are to be understood in particular as monoclonal antibodies.
  • the polypeptides according to the invention of the sequences Seq. ID Nos 77-85, 87, 88, 90, 91, 93, 95-108, 112-117, 119, 122, 124-126, 132, 133, 135, 137-160 and 179-209 can also be used as a tool for Finding drugs against breast cancer can be used, which is also the subject of the present invention.
  • the present invention also relates to the use of
  • Nucleic acid sequences according to the sequences Seq. ID No. 1 to Seq. ID No. 76 and Seq. ID No. 161 to Seq. ID No. 178 for the expression of polypeptides that can be used as tools for finding active substances against breast cancer.
  • the invention also relates to the use of the polypeptide partial sequences Seq found. ID No. 77 to Seq. ID No. 160 and Seq. ID No. 179 to seq. ID No. 209 as a drug in gene therapy for the treatment of breast cancer, or for the manufacture of a drug for the treatment of breast cancer.
  • the invention also relates to medicaments which have at least one polypeptide partial sequence Seq. ID No. 77 to Seq. ID No. 160 and Seq. ID No. 179 to seq. ID No. 209 included.
  • the nucleic acid sequences according to the invention found can also be genomic or mRNA sequences.
  • the invention also relates to genomic genes, their exon and intron structure and their splice variants, obtainable from the cDNAs of the sequences Seq. ID No. 1 to Seq. ID No. 76 and Seq. ID No. 161 to Seq. ID No. 178, and their use together with suitable regulatory elements, such as suitable promoters and / or enhancers.
  • suitable regulatory elements such as suitable promoters and / or enhancers.
  • BAC, PAC and cosmid clones isolated in this way are hybridized with the aid of fluorescence in situ hybridization to metaphase chromosomes and corresponding chromosome sections on which the corresponding genomic genes lie are identified.
  • BAC, PAC and cosmid clones are sequenced in order to elucidate the corresponding genomic genes in their complete structure (promoters, enhancers, silencers, exons and introns).
  • BAC, PAC and cosmid clones can be used as independent molecules for gene transfer (see FIG. 5).
  • the invention also relates to BAC, PAC and cosmid clones containing functional genes and their chromosomal localization, according to the sequences Seq. ID. No. 1 to Seq. ID No. 76 and Seq. ID No. 161 to Seq. ID No. 179, for use as a gene transfer vehicle.
  • nucleic acids nucleic acids are to be understood in the full invention: mRNA, partial cDNA, full length cDNA and genomic genes (chromosomes).
  • ORF Open Reading Frame, a defined sequence of amino acids that can be derived from the cDNA sequence.
  • Contig A lot of DNA sequences that are due to very large
  • Module domain of a protein with a defined sequence that represents a structural unit and occurs in different proteins
  • Fig. 1 shows the systematic gene search in the Incyte LifeSeq
  • Fig. 2b1-2b4 shows the entire principle of EST assembly.
  • Fig. 3 shows the in silico subtraction of gene expression in different tissues
  • 4a shows the determination of the tissue-specific expression via electronic Northern.
  • Figure 5 shows the isolation of BAC and PAC genomic clones.
  • the following examples explain the preparation of the nucleic acid sequences according to the invention without restricting the invention to these examples and nucleic acid sequences.
  • a partial DNA sequence S e.g. B. a single EST or a contig of ESTs are using a standard program for homology search, z. B. BLAST (Altschul, SF, Gish W., Miller, W., Myers, EW and Lipman, DJ (1990) J. Mol. Bio!., 215, 403-410), BLAST2 (Altschul, SF, Madden, T. L, Schaffer, AA, Zhang, J., Zhang, Z., Miller, W. and Lipman, DJ (1997) Nucleic Acids
  • tissue-specific occurrence frequencies of this partial sequence S are referred to as electronic Northern blot.
  • No. 42 found that occurs 12 times more in normal breast tissue than in tumor tissue.
  • Musculoskeletal system 0.0103 0.0120 0.85651.1675
  • Gastrointestinal 0, .0122
  • Musculoskeletal system 0.0240 0.0000 undef 0.0000
  • Musculoskeletal system 0.0086 0.0000 undef 0.0000
  • Musculoskeletal system 0.0274 0.0000 and 0.0000

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Genetics & Genomics (AREA)
  • Toxicology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne des séquences d'acides nucléiques humains, d'ARNm, d'ADNc, génomiques, provenant de tissus mammaires sains, qui codent pour des produits géniques ou des parties de ces derniers, ainsi que leur utilisation. L'invention concerne également les polypeptides pouvant être obtenus par l'intermédiaire de ces séquences, et leur utilisation.
PCT/DE1999/000909 1998-03-20 1999-03-19 Sequences d'acides nucleiques humains provenant de tissus mammaires sains WO1999047655A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2000536838A JP2002506639A (ja) 1998-03-20 1999-03-19 ヒト正常乳房組織からのヒト核酸配列
EP99924684A EP1070125A2 (fr) 1998-03-20 1999-03-19 Sequences d'acides nucleiques humains provenant de tissus mammaires sains

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19813835A DE19813835A1 (de) 1998-03-20 1998-03-20 Menschliche Nukleinsäuresequenzen aus Brustnormalgewebe
DE19813835.0 1998-03-20

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WO1999047655A2 true WO1999047655A2 (fr) 1999-09-23
WO1999047655A3 WO1999047655A3 (fr) 2000-07-06

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000029574A2 (fr) * 1998-11-18 2000-05-25 Incyte Pharmaceuticals, Inc. Genes associes a des inflammations
WO2000047752A2 (fr) * 1999-02-12 2000-08-17 University Of Iowa Research Foundation Diagnostics et therapies de la degenerescence maculaire
WO2001009178A2 (fr) * 1999-08-03 2001-02-08 Incyte Genomics, Inc. Proteines chaperons humaines
WO2001012823A2 (fr) * 1999-05-28 2001-02-22 University Of Iowa Research Foundation Diagnostic et traitement de la degenerescence maculaire
WO2002008260A2 (fr) * 2000-07-26 2002-01-31 Stanford University Proteine bstp-ecg1 et reactifs associes et procedes d'utilisation de ceux-ci
EP1179590A1 (fr) * 1999-04-30 2002-02-13 Kurokawa, Kiyoshi Proteine meg-3
US6458943B1 (en) 1998-02-17 2002-10-01 Institut National De La Sante Et De La Recherche Medicale hD54 polynucleotides
US7862819B2 (en) * 2001-02-23 2011-01-04 The J. David Gladstone Institutes Diacylglycerol O-acyltransferase 2α (DGAT2α)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050100998A1 (en) * 2000-08-21 2005-05-12 Prescott Stephen M. Human cytidine monophosphate (cmp) kinase cdna
WO2002044367A1 (fr) * 2000-11-29 2002-06-06 Takeda Chemical Industries, Ltd. Procede de criblage de medicaments contre le diabete
US8735082B2 (en) 2008-11-10 2014-05-27 The United States Of America, As Represented By The Secretary, Department Of Health And Human Services Gene signature for predicting prognosis of patients with solid tumors

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
BONFIELD, J. ET AL.: "A new sequence assembly program" NUCLEIC ACIDS RESEARCH, Bd. 23, 1995, Seiten 4992-4994, XP002127627 in der Anmeldung erw{hnt *
BRENNER, A.J. ET AL.: "Preferential loss of expression of p16INK4a rather than p19ARF in breast cancer" CLINICAL CANCER RESEARCH, Bd. 2, 1996, Seiten 1993-1998, XP002128956 *
Hillier, L. et al.:"WasU-NCI human EST Project":ab01h08.r1 Stratagene fetal retina 937202; Homo sapiens cDNA clone 839583 5' EMBL DATABASE ENTRY HS1289285;ACCESSION NUMBER AA489946, 02. Juli 1997 XP002128964 *
HUANG, R.-P. ET AL.: "Decreased Egr-1 expression in human, mouse and rat mamary cells and tissues correlates with tumour formation" INTERNATIONAL JOURNAL OF CANCER, Bd. 72, 1997, Seiten 102-109, XP002128955 *
Marra, M. et al.:"The WashU-HHMI Mouse EST Project":ms06e09.r1 Stratagene mouse skin; Mus musculus cDNA clone606184 5' EMBL DATABASE ENTRY MMAA145515; ACCESSION NUMBER AA145515, 14. Dezember 1996 XP002128962 *
Marra, M. et al.:"The WashU-HHMI Mouse EST Project":vq19c08.r1 Barstead stromal cell line MPLRB8; Mus musculus cDNA clone 1094702 5' EMBL DATABASE ENTRY AA869913; ACCESSION NUMBER AA869913, 17. M{rz 1998 XP002128963 *
Marra, M. et al.:"WashU-HHMI Mouse EST Project":vv29e04.r1 Stratagene mouse heart; Mus musculus cDNA clone 1223838 5'" EMBL DATABASE ENTRY AA667452; ACCESSION NUMBER AA667452, 20. November 1997 XP002128961 *
NISHIKIMI, A. ET AL.: "A novel mammalian nuclear protein similar to Schizosaccharomyces pombe Prp1/ Zer1p and Saccharomyces cerevisiae Prp6p pre-mRNA splicing factors" BIOCHIMICA BIOPHYSICA ACTA, Bd. 1435, 1999, Seiten 147-152, XP000872047 & Nishikimi et al. EMBL DATABASE ENTRY AB019219; ACCESSION NUMBER AB019219; 11. Januar 1999 *
SCHMITT, A.O. ET AL.: "Exhaustive mining of EST libraries for genes differentially expressed in normal and tumor tissue" NUCLEIC ACIDS RESEARCH, Bd. 27, Nr. 21, 1. November 1999 (1999-11-01), Seiten 4251-4260, XP002126641 *
SCHULER, G.D.: "Sequence mapping by electronic PCR" GENOME RESEARCH, Bd. 7, 1997, Seiten 541-550, XP002127628 in der Anmeldung erw{hnt *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6458943B1 (en) 1998-02-17 2002-10-01 Institut National De La Sante Et De La Recherche Medicale hD54 polynucleotides
WO2000029574A3 (fr) * 1998-11-18 2000-10-05 Incyte Pharma Inc Genes associes a des inflammations
WO2000029574A2 (fr) * 1998-11-18 2000-05-25 Incyte Pharmaceuticals, Inc. Genes associes a des inflammations
WO2000047752A2 (fr) * 1999-02-12 2000-08-17 University Of Iowa Research Foundation Diagnostics et therapies de la degenerescence maculaire
US6593104B1 (en) 1999-02-12 2003-07-15 University Of Iowa Research Foundation Macular degeneration diagnostics and therapeutics
WO2000047752A3 (fr) * 1999-02-12 2001-08-16 Univ Iowa Res Found Diagnostics et therapies de la degenerescence maculaire
US6417342B1 (en) 1999-02-12 2002-07-09 University Of Iowa Research Foundation Macular degeneration diagnostics and therapeutics
EP1179590A1 (fr) * 1999-04-30 2002-02-13 Kurokawa, Kiyoshi Proteine meg-3
EP1179590A4 (fr) * 1999-04-30 2002-09-18 Kurokawa Kiyoshi Proteine meg-3
WO2001012823A2 (fr) * 1999-05-28 2001-02-22 University Of Iowa Research Foundation Diagnostic et traitement de la degenerescence maculaire
WO2001012823A3 (fr) * 1999-05-28 2001-08-30 Univ Iowa Res Found Diagnostic et traitement de la degenerescence maculaire
WO2001009178A3 (fr) * 1999-08-03 2001-09-27 Incyte Genomics Inc Proteines chaperons humaines
WO2001009178A2 (fr) * 1999-08-03 2001-02-08 Incyte Genomics, Inc. Proteines chaperons humaines
WO2002008260A2 (fr) * 2000-07-26 2002-01-31 Stanford University Proteine bstp-ecg1 et reactifs associes et procedes d'utilisation de ceux-ci
WO2002008260A3 (fr) * 2000-07-26 2002-10-17 Univ Stanford Proteine bstp-ecg1 et reactifs associes et procedes d'utilisation de ceux-ci
US7862819B2 (en) * 2001-02-23 2011-01-04 The J. David Gladstone Institutes Diacylglycerol O-acyltransferase 2α (DGAT2α)

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WO1999047655A3 (fr) 2000-07-06
JP2002506639A (ja) 2002-03-05
EP1070125A2 (fr) 2001-01-24
DE19813835A1 (de) 1999-09-23

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