EP1414845A2 - Proteines humaines secretees - Google Patents

Proteines humaines secretees

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Publication number
EP1414845A2
EP1414845A2 EP02782476A EP02782476A EP1414845A2 EP 1414845 A2 EP1414845 A2 EP 1414845A2 EP 02782476 A EP02782476 A EP 02782476A EP 02782476 A EP02782476 A EP 02782476A EP 1414845 A2 EP1414845 A2 EP 1414845A2
Authority
EP
European Patent Office
Prior art keywords
seq
polypeptide
referenced
fragment
conesponding
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP02782476A
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German (de)
English (en)
Other versions
EP1414845A4 (fr
Inventor
Craig A. Rosen
Steven M. Ruben
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Human Genome Sciences Inc
Original Assignee
Human Genome Sciences Inc
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Filing date
Publication date
Application filed by Human Genome Sciences Inc filed Critical Human Genome Sciences Inc
Publication of EP1414845A2 publication Critical patent/EP1414845A2/fr
Publication of EP1414845A4 publication Critical patent/EP1414845A4/fr
Withdrawn legal-status Critical Current

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Classifications

    • 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
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • 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
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to human secreted proteins/polypeptides, and isolated nucleic acid molecules encoding said proteins/polypeptides, useful for detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating diabetes mellitus and conditions related thereto.
  • Antibodies that bind these polypeptides are also encompassed by the present invention.
  • vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies are also encompassed by the present invention.
  • the invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention.
  • the present invention further encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention.
  • IDDM Insulin-Dependent Diabetes Mellitus
  • NIDDM Non-Insulin-Dependent Diabetes Mellitus
  • insulin resistance eventually leads to the abolishment of insulin secretion resulting in insulin deficiency.
  • Insulin resistance at least in part, ensues from a block at the level of glucose uptake and phosphorylation in humans.
  • Diabetics demonstrate a decrease in expression in adipose tissue of insulin-receptor substrate 1 ("IRS1") (Carvalho et al., FASEB J 13(15):2173-8 (1999)), glucose transporter 4 ("GLUT4") (Garvey et al., Diabetes 41(4):465-75 (1992)), and the novel abundant protein M gene transcript 1 ("apMl”) (Statnick et al., Int I Exp Diabetes 1(2): 81-8 (2000)), as well as other as of yet unidentified factors. Insulin deficiency in NIDDM leads to failure of normal pancreatic beta-cell function and eventually to pancreatic-beta cell death.
  • Insulin affects fat, muscle, and liver. Insulin is the major regulator of energy metabolism. Malfunctioning of any step(s) in insulin secretion and/or action can lead to many disorders, including for example the dysregulation of oxygen utilization, adipogenesis, glycogenesis, lipogenesis, glucose uptake, protein synthesis, thermogenesis, and maintenance of the basal metabolic rate. This malfunctioning results in diseases and/or disorders that include, but are not limited to, hyperinsulinemia, insulin resistance, insulin deficiency, hyperglycemia, hyperlipidemia, hyperketonemia, and diabetes. Numerous debilitating diabetes-related secondary effects include, but are not limited to, obesity, forms of blindness (cataracts and diabetic retinopathy), limb amputations, kidney failure, fatty liver, coronary artery disease, and neuropathy.
  • Some of the current drugs used to treat insulin resistance and/or diabetes are inadequate due to the dosage amounts and frequency with which they have to be administered as a result of poor pharmacokinetic properties, the lack of effective control over blood sugar levels, and potential side effects, among other reasons.
  • Diabetes Therapeutic proteins in their native state or when recombinantly produced exhibit a rapid in vivo clearance. Typically, significant amounts of therapeutics are required to be effective during therapy.
  • small molecules smaller than the 20 kDa range can be readily filtered through the renal tubules (glomerulus) leading to dose-dependent nephrotoxicity. Therefore, there is a need for improvement in treatment (e.g., a need for prolonging the effects of therapeutics of diabetes and/or diabetes related conditions).
  • the present invention encompasses human secreted proteins/polypeptides, and isolated nucleic acid molecules encoding said proteins/polypeptides, useful for detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating diabetes mellitus and conditions related thereto.
  • Antibodies that bind these polypeptides are also encompassed by the present invention; as are vectors, host cells, and recombinant and synthetic methods for producing said polynucleotides, polypeptides, and/or antibodies.
  • the invention further encompasses screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention.
  • the present invention also encompasses methods and compositions for inhibiting or enhancing the production and function of the polypeptides of the present invention. Detailed Description Polynucleotides and Polypeptides of the Invention
  • Table 1A summarizes information concerning certain polypnucleotides and polypeptides of the invention.
  • the first column provides the gene number in the application for each clone identifier.
  • the second column provides a unique clone identifier, "Clone ID:”, for a cDNA clone related to each contig sequence disclosed in Table 1A.
  • Third column the cDNA Clones identified in the second column were deposited as indicated in the third column (i.e. by ATCC Deposit No:Z and deposit date). Some of the deposits contain multiple different clones corresponding to the same gene.
  • "Vector” refers to the type of vector contained in the corresponding cDNA Clone identified in the second column.
  • nucleotide sequence identified as "NT SEQ ED NO:X” was assembled from partially homologous ("overlapping") sequences obtained from the corresponding cDNA clone identified in the second column and, in some cases, from additional related cDNA clones.
  • the overlapping sequences were assembled into a single contiguous sequence of high redundancy (usually three to five overlapping sequences at each nucleotide position), resulting in a final sequence identified as SEQ ED NO:X.
  • 'Total NT Seq.” refers to the total number of nucleotides in the contig sequence identified as SEQ ED NO:X.”
  • the deposited clone may contain all or most of these sequences, reflected by the nucleotide position indicated as "5' NT of Clone Seq.” (seventh column) and the "3' NT of Clone Seq.” (eighth column) of SEQ ED NO:X.
  • the nucleotide position of SEQ ED NO:X of the putative start codon (methionine) is identified as "5' NT of Start Codon.”
  • the nucleotide position of SEQ ED NO:X of the predicted signal sequence is identified as "5' NT of First AA of Signal Pep.”
  • the translated amino acid sequence, beginning with the methionine is identified as "AA SEQ ED NO:Y,” although other reading frames can also be routinely translated using known molecular biology techniques. The polypeptides produced by these alternative open reading frames are specifically contemplated by the present invention.
  • the first and last amino acid position of SEQ ED NO:Y of the predicted signal peptide is identified as "First AA of Sig Pep" and "Last AA of Sig Pep.”
  • the predicted first amino acid position of SEQ ED NO: Y of the secreted portion is identified as "Predicted First AA of Secreted Portion”.
  • the amino acid position of SEQ ED NO:Y of the last amino acid encoded by the open reading frame is identified in the fifteenth column as "Last AA of ORF”.
  • SEQ ED NO:X (where X may be any of the polynucleotide sequences disclosed in the sequence listing) and the translated SEQ ED NO:Y (where Y may be any of the polypeptide sequences disclosed in the sequence listing) are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below.
  • SEQ ED NO:X is useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ED NO:X or the cDNA contained in the deposited clone. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention.
  • polypeptides identified from SEQ ED NO:Y may be used, for example, to generate antibodies which bind specifically to proteins containing the polypeptides and the secreted proteins encoded by the cDNA clones identified in Table 1A and/or elsewhere herein
  • DNA sequences generated by sequencing reactions can contain sequencing errors.
  • the errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence.
  • the erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence.
  • the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).
  • the present invention provides not only the generated nucleotide sequence identified as SEQ ED NO:X, and the predicted translated amino acid sequence identified as SEQ ED NO:Y, but also a sample of plasmid DNA containing a human cDNA of the invention deposited with the ATCC, as set forth in Table 1A.
  • the nucleotide sequence of each deposited plasmid can readily be determined by sequencing the deposited plasmid in accordance with known methods
  • amino acid sequence of the protein encoded by a particular plasmid can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence.
  • Table 1A Also provided in Table 1A is the name of the vector which contains the cDNA plasmid. Each vector is routinely used in the art. The following additional information is provided for convenience.
  • Vectors Lambda Zap U.S. Patent Nos. 5,128,256 and 5,286,636
  • Uni-Zap XR U.S.
  • Patent Nos. 5,128, 256 and 5,286,636 Zap Express (U.S. Patent Nos. 5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res. 76:7583-7600 (1988); Alting-Mees, M. A. and Short, I. M., Nucleic Acids Res. 17:9494 (1989)) and pBK (Alting-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La lolla, CA, 92037.
  • Phagemid pBS contains an ampicillin resistance gene and pBK contains a neomycin resistance gene.
  • Phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene
  • Vectors pSportl, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0 were obtained from Life Technologies, Inc., P. O. Box 6009, Gaithersburg, MD 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., et al., Focus 75:59 (1993). Vector lafmid BA (Bento Soares, Columbia University, New York, NY) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue.
  • Vector pCR ® 2.1 which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, CA 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 76:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).
  • the present invention also relates to the genes corresponding to SEQ ED NO:X, SEQ ED NO:Y, and/or a deposited cDNA (cDNA Clone ED).
  • the corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include, but are not limited to, preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material.
  • alielic variants, orthologs, and/or species homologs are also provided in the present invention. Procedures known in the art can be used to obtain full-length genes, alielic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of genes corresponding to SEQ ED NO:X and SEQ ED NO:Y using information from the sequences disclosed herein or the clones deposited with the ATCC.
  • alielic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for alielic variants and/or the desired homologue.
  • the present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ ED NO:X and/or a cDNA contained in ATCC Deposit No.Z.
  • the present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ ED NO:Y, a polypeptide encoded by SEQ ED NO:X, and/or a polypeptide encoded by a cDNA contained in ATCC deposit No.Z.
  • Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ ED NO: Y, a polypeptide encoded by SEQ ED NO:X and/or a polypeptide encoded by the cDNA contained in ATCC Deposit No.Z, are also encompassed by the invention.
  • the present invention further encompasses a polynucleotide comprising, or alternatively consisting of the complement of the nucleic acid sequence of SEQ ED NO:X, and/or the complement of the coding strand of the cDNA contained in ATCC Deposit No.Z. Description of Table IB (Comprised of Tables IB.l and 1B.2)
  • Table IB.l and Table 1B.2 summarize some of the polynucleotides encompassed by the invention (including cDNA clones related to the sequences (Clone ED:), contig sequences (contig identifier (Contig ED:) and contig nucleotide sequence identifiers (SEQ ED NO:X)) and further summarizes certain characteristics of these polynucleotides and the polypeptides encoded thereby.
  • the first column of Tables IB.l and 1B.2 provide the gene numbers in the application for each clone identifier.
  • the second column of Tables IB.l and 1B.2 provide unique clone identifiers, "Clone ED:”, for cDNA clones related to each contig sequence disclosed in Table 1A and/or Table IB.
  • the third column of Tables IB.l and 1B.2 provide unique contig identifiers, "Contig ED:” for each of the contig sequences disclosed in these tables.
  • the fourth column of Tables IB.l and 1B.2 provide the sequence identifiers, "SEQ ID NO:X", for each of the contig sequences disclosed in Table 1A and/or IB.
  • Table IB.l The fifth column of Table IB.l, "ORF (From-To)", provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ ED NO:X that delineates the preferred open reading frame (ORF) that encodes the amino acid sequence shown in the sequence listing and referenced in Table IB.l as SEQ ED NO:Y (column 6).
  • Column 7 of Table IB.l lists residues comprising predicted epitopes contained in the polypeptides encoded by each of the preferred ORFs (SEQ ID NO:Y).
  • polypeptides of the invention comprise, or alternatively consist of, one, two, three, four, five or more of the predicted epitopes described in Table IB.l. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly.
  • Column 8 of Table IB.l (“Cytologic Band") provides the chromosomal location of polynucleotides corresponding to SEQ ED NO:X. Chromosomal location was determined by finding exact matches to EST and cDNA sequences contained in the NCBI (National Center for Biotechnology Information) UniGene database.
  • OMEM identification number is disclosed in Table IB.l, column 9 labeled "OMEM Disease Reference(s)". A key to the OMEM reference identification numbers is provided in Table 5.
  • Table IB.2 'Tissue Distribution” shows the expression profile of tissue, cells, and/or cell line libraries which express the polynucleotides of the invention.
  • the first code number shown in Table 1B.2 column 5 represents the tissue/cell source identifier code corresponding to the key provided in Table 4. Expression of these polynucleotides was not observed in the other tissues and/or cell libraries tested.
  • the second number in column 5 represents the number of times a sequence corresponding to the reference polynucleotide sequence (e.g., SEQ ED NO:X) was identified in the corresponding tissue/cell source.
  • tissue/cell source identifier codes in which the first two letters are "AR" designate information generated using DNA array technology.
  • cDNAs were amplified by PCR and then transferred, in duplicate, onto the array. Gene expression was assayed through hybridization of first strand cDNA probes to the DNA array. cDNA probes were generated from total RNA extracted from a variety of different tissues and cell lines. Probe synthesis was performed in the presence of 33 P dCTP, using oligo(dT) to prime reverse transcription. After hybridization, high stringency washing conditions were employed to remove non-specific hybrids from the array. The remaining signal, emanating from each gene target, was measured using a Phosphorimager.
  • Phosphor Stimulating Luminescence which reflects the level of phosphor signal generated from the probe hybridized to each of the gene targets represented on the array.
  • a local background signal subtraction was performed before the total signal generated from each array was used to normalize gene expression between the different hybridizations.
  • the value presented after "[array code]:” represents the mean of the duplicate values, following background subtraction and probe normalization.
  • One of skill in the art could routinely use this information to identify normal and/or diseased tissue(s) which show a predominant expression pattern of the corresponding polynucleotide of the invention or to identify polynucleotides which show predominant and/or specific tissue and/or cell expression.
  • the first column provides a unique clone identifier, "Clone ED:”, for a cDNA clone related to each contig sequence.
  • the second column provides the sequence identifier, "SEQ ID NO:X”, for each contig sequence.
  • the third column provides a unique contig identifier, "Contig ED:” for each contig sequence.
  • the fourth column provides a BAC identifier "BAC ED NO:A” for the BAC clone referenced in the corresponding row of the table.
  • the fifth column provides the nucleotide sequence identifier, "SEQ ED NO:B” for a fragment of the BAC clone identified in column four of the corresponding row of the table.
  • the sixth column "Exon From- To" provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ ID NO:B which delineate certain polynucleotides of the invention that are also exemplary members of polynucleotide sequences that encode polypeptides of the invention (e.g., polypeptides containing amino acid sequences encoded by the polynucleotide sequences delineated in column six, and fragments and variants thereof).
  • the present invention encompasses a method of detecting, preventing, diagnosing, prognosticating, treating, and or ameliorating diabetes mellitus; comprising administering to a patient in which such treatment, prevention, or amelioration is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) represented by Table 1A, Table IB, and Table 1C, in an amount effective to detect, prevent, diagnose, prognosticate, treat, and/or ameliorate the disease or disorder.
  • the polynucleotides, polypeptides, agonists, or antagonists of the present invention can be used in assays to test for one or more biological activities. If these polynucleotides and polypeptides do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the polynucleotides or polypeptides, or agonists or antagonists thereof (including antibodies) could be used to treat the associated disease.
  • Table ID provides information related to biological activities for polynucleotides and polypeptides of the invention (including antibodies, agonists, and or antagonists thereof).
  • Table ID also provides information related to assays which may be used to test polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) for the corresponding biological activities.
  • the first column (“Gene No.”) provides the gene number in the application for each clone identifier.
  • the second column (“cDNA Clone ID:”) provides the unique clone identifier for each clone as previously described and indicated in Tables 1A, IB, and lC.
  • the third column (“AA SEQ ED NO:Y”) indicates the Sequence Listing SEQ ED Number for polypeptide sequences encoded by the corresponding cDNA clones (also as indicated in Tables 1A, IB, and 2).
  • the fourth column (“Biological Activity”) indicates a biological activity corresponding to the indicated polypeptides (or polynucleotides encoding said polypeptides).
  • the fifth column (“Exemplary Activity Assay”) further describes the corresponding biological activity and provides information pertaining to the various types of assays which may be performed to test, demonstrate, or quantify the corresponding biological activity.
  • Table ID describes the use of FMAT technology, inter alia, for testing or demonstrating various biological activities.
  • Fluorometric microvolume assay technology (FMAT) is a fluorescence-based system which provides a means to perform nonradioactive cell- and bead-based assays to detect activation of cell signal transduction pathways. This technology was designed specifically for ligand binding and immunological assays.
  • FMAT technology may be used for peptide ligand binding assays, immunofluorescence, apoptosis, cytotoxicity, and bead-based immunocapture assays. See, Miraglia S et. al., "Homogeneous cell and bead based assays for highthroughput screening using flourometric microvolume assay technology," Journal of Biomolecular Screening; 4:193-204 (1999).
  • FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides (including polypeptide fragments and variants) to activate signal transduction pathways.
  • FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides to upregulate production of immunomodulatory proteins (such as, for example, interleukins, GM-CSF, Rantes, and Tumor Necrosis factors, as well as other cellular regulators (e.g. insulin)).
  • immunomodulatory proteins such as, for example, interleukins, GM-CSF, Rantes, and Tumor Necrosis factors, as well as other cellular regulators (e.g. insulin)
  • Table ID also describes the use of kinase assays for testing, demonstrating, or quantifying biological activity.
  • the phosphorylation and de-phosphorylation of specific amino acid residues e.g. Tyrosine, Serine, Threonine
  • cell-signal transduction proteins provides a fast, reversible means for activation and de-activation of cellular signal transduction pathways.
  • cell signal transduction via phosphorylation/de-phosphorylation is crucial to the regulation of a wide variety of cellular processes (e.g. proliferation, differentiation, migration, apoptosis, etc.).
  • kinase assays provide a powerful tool useful for testing, confirming, and/or identifying polypeptides (including polypeptide fragments and variants) that mediate cell signal transduction events via protein phosphorylation. See e.g., Forrer, P., Tamaskovic R., and Jaussi, R. "Enzyme-Linked Immunosorbent Assay for Measurement of JNK, ERK, and p38 Kinase Activities" Biol. Chem. 379(8-9): 1101-1110 (1998).
  • Polynucleotides encoding polypeptides of the present invention can be used in assays to test for one or more biological activities.
  • One such biological activity which may be tested includes the ability of polynucleotides and polypeptides of the invention to stimulate up-regulation or down-regulation of expression of particular genes and proteins.
  • polynucleotides and polypeptides of the present invention exhibit activity in altering particular gene and protein expression patterns, it is likely that these polynucleotides and polypeptides of the present invention may be involved in, or capable of effecting changes in, diseases associated with the altered gene and protein expression profiles.
  • polynucleotides, polypeptides, or antibodies of the present invention could be used to treat said associated diseases.
  • TaqMan® assays may be performed to assess the ability of polynucleotides (and polypeptides they encode) to alter the expression pattern of particular "target" genes.
  • TaqMan® reactions are performed to evaluate the ability of a test agent to induce or repress expression of specific genes in different cell types.
  • TaqMan® gene expression quantification assays (“TaqMan® assays") are well known to, and routinely performed by, those of ordinary skill in the art.
  • TaqMan® assays are performed in a two step reverse transcription / polymerase chain reaction (RT-PCR). In the first (RT) step, cDNA is reverse transcribed from total RNA samples using random hexamer primers. In the second (PCR) step, PCR products are synthesized from the cDNA using gene specific primers.
  • the Taqman® PCR reaction exploits the 5' nuclease activity of AmpliTaq Gold DNA Polymerase to cleave a Taqman® probe (distinct from the primers) during PCR.
  • the Taqman® probe contains a reporter dye at the 5 '-end of the probe and a quencher dye at the 3' end of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in suppression of the reporter fluorescence.
  • the probe specifically anneals between the forward and reverse primer sites.
  • AmpliTaq Fold DNA Polymerase then cleaves the probe between the reporter and quencher when the probe hybridizes to the target, resulting in increased fluorescence of the reporter (see Figure 2). Accumulation of PCR products is detected directly by monitoring the increase in fluorescence of the reporter dye.
  • probe fragments are displaced from the target, polymerization of the strand continues.
  • the 3'-end of the probe is blocked to prevent extension of the probe during PCR. This process occurs in every cycle and does not interfere with the exponential accumulation of product.
  • the increase in fluorescence signal is detected only if the target sequence is complementary to the probe and is amplified during PCR. Because of these requirements, any nonspecific amplification is not detected.
  • vector controls or constructs containing the coding sequence for the gene of interest are transfected into cells, such as for example 293T cells, and supernatants collected after 48 hours.
  • multiple primary human cells or human cell lines are used; such cells may include but are not limited to, Normal Human Dermal Fibroblasts, Aortic Smooth Muscle, Human Umbilical Vein Endothelial Cells, He ⁇ G2, Daudi, urkat, U937, Caco, and THP-1 cell lines.
  • Cells are plated in growth media and growth is arrested by culturing without media change for 3 days, or by switching cells to low serum media and incubating overnight. Cells are treated for 1, 6, or 24 hours with either vector control supernatant or sample supernatant (or purified/partially purified protein preparations in buffer).
  • Total RNA is isolated; for example, by using Trizol extraction or by using the Ambion RNAqueous(TM)-4PCR RNA isolation system. Expression levels of multiple genes are analyzed using TAQMAN, and expression in the test sample is compared to control vector samples to identify genes induced or repressed.
  • Table IE indicates particular disease classes and preferred indications for which polynucleotides, polypeptides, or antibodies of the present invention may be used in detecting, diagnosing, preventing, treating and/or ameliorating said diseases and disorders based on "target" gene expression patterns which may be up- or down-regulated by polynucleotides (and the encoded polypeptides) corresponding to each indicated cDNA Clone ID (shown in Table IE, Column 2).
  • the present invention encompasses a method of detecting, diagnosing, preventing, treating, and/or ameliorating a disease or disorder listed in the "Disease Class" and/or "Preferred Indication” columns of Table IE; comprising administering to a patient in which such detection, diagnosis, prevention, or treatment is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to detect, diagnose, prevent, treat, or ameliorate the disease or disorder.
  • the first and second columns of Table ID show the "Gene No.” and "cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in detecting, diagnosing, preventing, treating, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in the "Disease Class” or "Preferred Indication” Columns of Table IE.
  • the present invention also encompasses methods of detecting, diagnosing, preventing, treating, or ameliorating a disease or disorder listed in the "Disease Class” or "Preferred Indication” Columns of Table IE; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in the "Disease Class” or “Preferred Indication” Columns of Table IE.
  • the "Disease Class” Column of Table IE provides a categorized descriptive heading for diseases, disorders, and/or conditions (more fully described below) that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).
  • the "Preferred Indication" Column of Table lE describes diseases, disorders, and/or conditions that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).
  • Cell Line and “Exemplary Targets” Columns of Table IE indicate particular cell lines and target genes, respectively, which may show altered gene expression patterns (i.e., up- or down-regulation of the indicated target gene) in Taqman assays, performed as described above, utilizing polynucleotides of the cDNA Clone ID shown in the corresponding row. Alteration of expression patterns of the indicated “Exemplary Target” genes is correlated with a particular "Disease Class” and/or "Preferred Indication” as shown in the corresponding row under the respective column headings.
  • the "Exemplary Accessions” Column indicates GenBank Accessions (available online through the National Center for Biotechnology Information (NCBI) at http://www.ncbi.nlm.nih.gov/) which correspond to the "Exemplary Targets” shown in the adjacent row.
  • the recitation of "Cancer” in the “Disease Class” Column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate neoplastic diseases and/or disorders (e.g., leukemias, cancers, etc., as described below under “Hyperproliferative Disorders”).
  • the recitation of “Immune” in the "Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under "Hyperproliferative Disorders"), blood disorders (e.g., as described below under "Immune Activity” "Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under "Infectious Disease”).
  • neoplastic diseases e.g., as described below under "Hyperproliferative Disorders”
  • blood disorders e.g., as described below under “Immune Activity” "Cardiovascular Disorders” and/or “Blood-Related Disorders”
  • infections e.g., as described below under "
  • Angiogenesis in the "Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under "Hyperproliferative Disorders"), diseases and/or disorders of the cardiovascular system (e.g., as described below under "Cardiovascular Disorders"), diseases and/or disorders involving cellular and genetic abnormalities (e.g., as described below under "Diseases at the Cellular Level"), diseases and/or disorders involving angiogenesis (e.g., as described below under "Anti-Angiogenesis Activity”), to promote or inhibit cell or tissue regeneration (e.g., as described below under “Regeneration”), or to promote wound healing (e.g., as described below under "Wound Healing and Epithelial Cell Proliferation”).
  • neoplastic diseases
  • Diabetes in the "Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diabetes (including diabetes mellitus types I and II), as well as diseases and/or disorders associated with, or consequential to, diabetes (e.g. as described below under “Endocrine Disorders,” “Renal Disorders,” and “Gastrointestinal Disorders”).
  • Table 2 summarizes homology and features of some of the polypeptides of the invention.
  • the first column provides a unique clone identifier, "Clone ED:”, corresponding to a cDNA clone disclosed in Table 1A or Table IB.
  • the second column provides the unique contig identifier, "Contig ED:” corresponding to contigs in Table IB and allowing for correlation with the information in Table IB.
  • the third column provides the sequence identifier, "SEQ ED NO:X”, for the contig polynucleotide sequence.
  • the fourth column provides the analysis method by which the homology/identity disclosed in the Table was determined.
  • NR non-redundant protein database
  • PFAM protein families
  • polypeptides of the invention comprise, or alternatively consist of, an amino acid sequence encoded by a polynucleotide in SEQ ED NO:X as delineated in columns 8 and 9, or fragments or variants thereof.
  • Table 3 provides polynucleotide sequences that may be disclaimed according to certain embodiments of the invention.
  • the first column provides a unique clone identifier, "Clone ED”, for a cDNA clone related to contig sequences disclosed in Table IB.
  • the second column provides the sequence identifier, "SEQ ED NO:X”, for contig sequences disclosed in Table 1A and/or Table IB.
  • the third column provides the unique contig identifier, "Contig ED:”, for contigs disclosed in Table IB.
  • the fourth column provides a unique integer 'a' where 'a' is any integer between 1 and the final nucleotide minus 15 of SEQ ED NO:X
  • the fifth column provides a unique integer 'b' where 'b' is any integer between 15 and the final nucleotide of SEQ ED NO:X, where both a and b correspond to the positions of nucleotide residues shown in SEQ ED NO:X, and where b is greater than or equal to a + 14.
  • the uniquely defined integers can be substituted into the general formula of a-b, and used to describe polynucleotides which may be preferably excluded from the invention.
  • preferably excluded from the invention are at least one, two, three, four, five, ten, or more of the polynucleotide sequence(s) having the accession number(s) disclosed in the sixth column of this Table (including for example, published sequence in connection with a particular BAC clone).
  • preferably excluded from the invention are the specific polynucleotide sequence(s) contained in the clones corresponding to at least one, two, three, four, five, ten, or more of the available material having the accession numbers identified in the sixth column of this Table (including for example, the actual sequence contained in an identified BAC clone).
  • Table 4 provides a key to the tissue/cell source identifier code disclosed in Table 1B.2, column 5.
  • Column 1 provides the tissue/cell source identifier code disclosed in Table 1B.2, Column 5.
  • Columns 2-5 provide a description of the tissue or cell source. Note that “Description” and “Tissue” sources (i.e. columns 2 and 3) having the prefix “a_” indicates organs, tissues, or cells derived from “adult” sources. Codes corresponding to diseased tissues are indicated in column 6 with the word “disease.” The use of the word “disease” in column 6 is non-limiting.
  • the tissue or cell source may be specific (e.g.
  • tissue/cell source is a library
  • column 7 identifies the vector used to generate the library.
  • Table 5 provides a key to the OMEM reference identification numbers disclosed in Table IB.l, column 9.
  • OMIM reference identification numbers (Column 1) were derived from Online Mendelian Inheritance in Man (Online Mendelian Inheritance in Man, OMEM. McKusick- Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, MD) and National Center for Biotechnology Information, National Library of Medicine, (Bethesda, MD) 2000. World Wide Web URL: http://www.ncbi.nlm.nih.gov/omim/).
  • Column 2 provides diseases associated with the cytologic band disclosed in Table IB.l, column 8, as determined using the Morbid Map database.
  • Table 6 summarizes some of the ATCC Deposits, Deposit dates, and ATCC designation numbers of deposits made with the ATCC in connection with the present application. These deposits were made in addition to those described in the Table 1A.
  • Table 7 shows the cDNA libraries sequenced, and ATCC designation numbers and vector information relating to these cDNA libraries.
  • the first column shows the first four letters indicating the Library from which each library clone was derived.
  • the second column indicates the catalogued tissue description for the corresponding libraries.
  • the third column indicates the vector containing the corresponding clones.
  • the fourth column shows the ATCC deposit designation for each libray clone as indicated by the deposit information in Table 6.
  • isolated refers to material removed from its original environment (e.g., the natural environment if it is naturally occurring), and thus is altered “by the hand of man” from its natural state.
  • an isolated polynucleotide could be part of a vector or a composition of matter, or could be contained within a cell, and still be “isolated” because that vector, composition of matter, or particular cell is not the original environment of the polynucleotide.
  • isolated does not refer to genomic or cDNA libraries, whole cell total or mRNA preparations, genomic DNA preparations (including those separated by electrophoresis and transferred onto blots), sheared whole cell genomic DNA preparations or other compositions where the art demonstrates no distinguishing features of the polynucleotide/sequences of the present invention.
  • a "secreted" protein refers to those proteins capable of being directed to the ER, secretory vesicles, or the extracellular space as a result of a signal sequence, as well as those proteins released into the extracellular space without necessarily containing a signal sequence. If the secreted protein is released into the extracellular space, the secreted protein can undergo extracellular processing to produce a "mature" protein. Release into the extracellular space can occur by many mechanisms, including exocytosis and proteolytic cleavage.
  • a "polynucleotide” refers to a molecule having a nucleic acid sequence encoding SEQ ED NO:Y or a fragment or variant thereof (e.g., the polypeptide delinated in columns fourteen and fifteen of Table 1 A); a nucleic acid sequence contained in SEQ ED NO:X (as described in column 5 of Table 1A and/or column 3 of Table IB) or the complement thereof; a cDNA sequence contained in Clone ED: (as described in column 2 of Table 1A and/or Table IB and contained within a library deposited with the ATCC); a nucleotide sequence encoding the polypeptide encoded by a nucleotide sequence in SEQ ED NO:B as defined in column 6 (EXON From-To) of Table IC or a fragment or variant thereof; or a nucleotide coding sequence in SEQ ED NO:B as defined in column 6 of Table IC or the complement thereof.
  • the polynucleotide can contain the nucleotide sequence of the full length cDNA sequence, including the 5' and 3' untranslated sequences, the coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence.
  • a "polypeptide” refers to a molecule having an amino acid sequence encoded by a polynucleotide of the invention as broadly defined (obviously excluding poly-Phenylalanine or poly-Lysine peptide sequences which result from translation of a polyA tail of a sequence corresponding to a cDNA).
  • SEQ ED NO:X was often generated by overlapping sequences contained in multiple clones (contig analysis).
  • a representative clone containing all or most of the sequence for SEQ ID NO:X is deposited at Human Genome Sciences, Inc. (HGS) in a catalogued and archived library.
  • HGS Human Genome Sciences, Inc.
  • each clone is identified by a cDNA Clone ED (identifier generally referred to herein as Clone ED:).
  • Clone ED identifier generally referred to herein as Clone ED:
  • Each Clone ED is unique to an individual clone and the Clone ID is all the information needed to retrieve a given clone from the HGS library.
  • Table 7 provides a list of the deposited cDNA libraries.
  • Table 7 lists the deposited cDNA libraries by name and links each library to an ATCC Deposit. Library names contain four characters, for example, "HTWE.” The name of a cDNA clone (Clone ED) isolated from that library begins with the same four characters, for example "HTWEP07".
  • Table 1A and/or Table IB correlates the Clone ED names with SEQ ID NO:X. Thus, starting with an SEQ ED NO:X, one can use Tables 1A, IB, 6, 7, and 9 to determine the corresponding Clone ED, which library it came from and which ATCC deposit the library is contained in.
  • the polynucleotides of the invention are at least 15, at least
  • polynucleotides of the invention comprise a portion of the coding sequences, as disclosed herein, but do not comprise all or a portion of any intron.
  • the polynucleotides comprising coding sequences do not contain coding sequences of a genomic flanking gene (i.e., 5' or 3' to the gene of interest in the genome). In other embodiments, the polynucleotides of the invention do not contain the coding sequence of more than 1000, 500, 250, 100, 50, 25, 20, 15, 10, 5, 4, 3, 2, or 1 genomic flanking gene(s).
  • a "polynucleotide” of the present invention also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ ED NO:X, or the complement thereof (e.g., the complement of any one, two, three, four, or more of the polynucleotide fragments described herein), the polynucleotide sequence delineated in columns 7 and 8 of Table 1A or the complement thereof, the polynucleotide sequence delineated in columns 8 and 9 of Table 2 or the complement thereof, and/or cDNA sequences contained in Clone ED: (e.g., the complement of any one, two, three, four, or more of the polynucleotide fragments, or the cDNA clone within the pool of cDNA clones deposited with the ATCC, described herein), and/or the polynucleotide sequence delineated in column 6 of Table IC or the complement thereof.
  • SEQ ED NO:X or
  • “Stringent hybridization conditions” refers to an overnight incubation at 42 degree C in a solution comprising 50% formamide, 5x SSC (750 mM NaCl, 75 mM trisodium citrate), 50 mM sodium phosphate (pH 7.6), 5x Denhardt's solution, 10% dextran sulfate, and 20 ⁇ g/ml denatured, sheared salmon sperm DNA, followed by washing the filters in O.lx SSC at about 65 degree C.
  • nucleic acid molecules that hybridize to the polynucleotides of the present invention at lower stringency hybridization conditions. Changes in the stringency of hybridization and signal detection are primarily accomplished through the manipulation of formamide concentration (lower percentages of formamide result in lowered stringency); salt conditions, or temperature.
  • washes performed following stringent hybridization can be done at higher salt concentrations (e.g. 5X SSC).
  • blocking reagents include Denhardt's reagent, BLOTTO, heparin, denatured salmon sperm DNA, and commercially available proprietary formulations.
  • the inclusion of specific blocking reagents may require modification of the hybridization conditions described above, due to problems with compatibility.
  • polynucleotide which hybridizes only to polyA ⁇ sequences (such as any 3' terminal polyA+ tract of a cDNA shown in the sequence listing), or to a complementary stretch of T (or U) residues, would not be included in the definition of "polynucleotide,” since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (e.g., practically any double-stranded cDNA clone generated using oligo dT as a primer).
  • polynucleotide of the present invention can be composed of any polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA.
  • polynucleotides can be composed of single- and double- stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double- stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions.
  • polynucleotide can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA.
  • a polynucleotide may also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons.
  • Modified bases include, for example, tritylated bases and unusual bases such as inosine.
  • polynucleotide embraces chemically, enzymatically, or metabolically modified forms.
  • the polynucleotides of the invention are at least 15, at least
  • polynucleotides of the invention comprise a portion of the coding sequences, as disclosed herein, but do not comprise all or a portion of any intron.
  • the polynucleotides comprising coding sequences do not contain coding sequences of a genomic flanking gene (i.e., 5' or 3' to the gene of interest in the genome).
  • the polynucleotides of the invention do not contain the coding sequence of more than 1000, 500, 250, 100, 50, 25, 20, 15, 10, 5, 4, 3, 2, or 1 genomic flanking gene(s).
  • SEQ ED NO:X refers to a polynucleotide sequence described in column 5 of Table 1A
  • SEQ ED NO:Y refers to a polypeptide sequence described in column 10 of Table 1A
  • SEQ ED NO:X is identified by an integer specified in column 6 of Table 1A.
  • the polypeptide sequence SEQ ED NO:Y is a translated open reading frame (ORF) encoded by polynucleotide SEQ ED NO:X.
  • the polynucleotide sequences are shown in the sequence listing immediately followed by all of the polypeptide sequences.
  • a polypeptide sequence corresponding to polynucleotide sequence SEQ ED NO: 2 is the first polypeptide sequence shown in the sequence listing.
  • the second polypeptide sequence corresponds to the polynucleotide sequence shown as SEQ ED NO:3, and so on.
  • the polypeptide of the present invention can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids.
  • the polypeptides may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini.
  • polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods.
  • Modifications include acetylation, acylation, ADP- ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination.
  • SEQ ED NO:X refers to a polynucleotide sequence described, for example, in Tables 1A, Table IB, or Table 2, while “SEQ ED NO:Y” refers to a polypeptide sequence described in column 11 of Table 1A and or of Table IB. SEQ ED NO:X is identified by an integer specified in column 4 of Table IB.
  • the polypeptide sequence SEQ ID NO:Y is a translated open reading frame (ORF) encoded by polynucleotide SEQ ED NO:X.
  • Clone ED: refers to a cDNA clone described in column 2 of Table 1A and/or IB.
  • a polypeptide having functional activity refers to a polypeptide capable of displaying one or more known functional activities associated with a full-length (complete) protein. Such functional activities include, but are not limited to, biological activity (e.g. activity useful in treating, preventing and/or ameliorating diabetes mellitus), antigenicity (ability to bind [or compete with a polypeptide for binding] to an anti-polypeptide antibody), immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide.
  • the polypeptides of the invention can be assayed for functional activity (e.g. biological activity) using or routinely modifying assays known in the art, as well as assays described herein.
  • a polypeptide having biological activity refers to a polypeptide exhibiting activity similar to, but not necessarily identical to, an activity of a polypeptide of the present invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. In the case where dose dependency does exist, it need not be identical to that of the polypeptide, but rather substantially similar to the dose-dependence in a given activity as compared to the polypeptide of the present invention (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the polypeptide of the present invention).
  • Table 1A summarizes information concerning certain polypnucleotides and polypeptides of the invention.
  • the first column provides the gene number in the application for each clone identifier.
  • the second column provides a unique clone identifier, "Clone ED:”, for a cDNA clone related to each contig sequence disclosed in Table 1A.
  • Third column the cDNA Clones identified in the second column were deposited as indicated in the third column (i.e. by ATCC Deposit No:Z and deposit date). Some of the deposits contain multiple different clones corresponding to the same gene.
  • “Vector” refers to the type of vector contained in the corresponding cDNA Clone identified in the second column.
  • nucleotide sequence identified as "NT SEQ ID NO:X” was assembled from partially homologous ("overlapping") sequences obtained from the corresponding cDNA clone identified in the second column and, in some cases, from additional related cDNA clones.
  • the overlapping sequences were assembled into a single contiguous sequence of high redundancy (usually three to five overlapping sequences at each nucleotide position), resulting in a final sequence identified as SEQ ED NO:X.
  • Total NT Seq refers to the total number of nucleotides in the contig sequence identified as SEQ ED NO:X.”
  • the deposited clone may contain all or most of these sequences, reflected by the nucleotide position indicated as "5' NT of Clone Seq.” (seventh column) and the "3' NT of Clone Seq.” (eighth column) of SEQ ID NO:X.
  • the nucleotide position of SEQ ED NO:X of the putative start codon (methionine) is identified as "5' NT of Start Codon.”
  • the nucleotide position of SEQ ED NO:X of the predicted signal sequence is identified as "5' NT of First AA of Signal Pep.”
  • the translated amino acid sequence, beginning with the methionine is identified as "AA SEQ ID NO:Y,” although other reading frames can also be routinely translated using known molecular biology techniques. The polypeptides produced by these alternative open reading frames are specifically contemplated by the present invention.
  • the first and last amino acid position of SEQ ED NO:Y of the predicted signal peptide is identified as "First AA of Sig Pep" and "Last AA of Sig Pep.”
  • the predicted first amino acid position of SEQ ID NO:Y of the secreted portion is identified as "Predicted First AA of Secreted Portion”.
  • the amino acid position of SEQ ED NO:Y of the last amino acid encoded by the open reading frame is identified in the fifteenth column as "Last AA of ORF'.
  • SEQ ED NO:X (where X may be any of the polynucleotide sequences disclosed in the sequence listing) and the translated SEQ ED NO:Y (where Y may be any of the polypeptide sequences disclosed in the sequence listing) are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below.
  • SEQ ED NO:X is useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ED NO:X or the cDNA contained in the deposited clone. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention.
  • polypeptides identified from SEQ ED NO:Y may be used, for example, to generate antibodies which bind specifically to proteins containing the polypeptides and the secreted proteins encoded by the cDNA clones identified in Table 1A and/or elsewhere herein Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence.
  • the present invention provides not only the generated nucleotide sequence identified as SEQ ED NO:X, and the predicted translated amino acid sequence identified as SEQ ED NO: Y, but also a sample of plasmid DNA containing a human cDNA of the invention deposited with the ATCC, as set forth in Table 1A.
  • the nucleotide sequence of each deposited plasmid can readily be determined by sequencing the deposited plasmid in accordance with known methods
  • amino acid sequence of the protein encoded by a particular plasmid can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence.
  • Table 1A Also provided in Table 1A is the name of the vector which contains the cDNA plasmid. Each vector is routinely used in the art. The following additional information is provided for convenience.
  • phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene
  • Vectors pSportl, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0 were obtained from Life Technologies, Inc., P. O. Box 6009, Gaithersburg, MD 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., et al., Focus 75:59 (1993). Vector lafmid BA (Bento Soares, Columbia University, New York, NY) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue.
  • Vector pCR ® 2.1 which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, CA 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 76:9677-9686 (1988) and Mead, D. et al, Bio/Technology 9: (1991).
  • the present invention also relates to the genes corresponding to SEQ ED NO:X, SEQ
  • cDNA Clone ED a deposited cDNA
  • the corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include, but are not limited to, preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material.
  • alielic variants, orthologs, and or species homologs are also provided in the present invention.
  • Procedures known in the art can be used to obtain full-length genes, alielic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of genes corresponding to SEQ ED NO:X and SEQ ID NO:Y using information from the sequences disclosed herein or the clones deposited with the ATCC.
  • alielic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for alielic variants and or the desired homologue.
  • the present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ ID NO:X and/or a cDNA contained in ATCC Deposit No.Z.
  • the present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ ED NO:Y, a polypeptide encoded by SEQ ED NO:X, and/or a polypeptide encoded by a cDNA contained in ATCC deposit No.Z.
  • Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ ED NO: Y, a polypeptide encoded by SEQ ED NO:X and/or a polypeptide encoded by the cDNA contained in ATCC Deposit No.Z, are also encompassed by the invention.
  • the present invention further encompasses a polynucleotide comprising, or alternatively consisting of the complement of the nucleic acid sequence of SEQ ED NO:X, and/or the complement of the coding strand of the cDNA contained in ATCC Deposit No.Z.
  • the first column in Table IB.l and Table 1B.2 provides the gene number in the application corresponding to the clone identifier.
  • the second column in Table IB.l and Table 1B.2 provides a unique "Clone ID:" for the cDNA clone related to each contig sequence disclosed in Table IB.l and Table IB.2.
  • This clone ED references the cDNA clone which contains at least the 5' most sequence of the assembled contig and at least a portion of SEQ ED NO:X as determined by directly sequencing the referenced clone.
  • the referenced clone may have more sequence than described in the sequence listing or the clone may have less.
  • a full-length cDNA can be obtained by methods described elsewhere herein.
  • the third column in Table IB.l and Table IB.2 provides a unique "Contig ED" identification for each contig sequence.
  • the fourth column in Table IB.l and Table 1B.2 provides the "SEQ ED NO:" identifier for each of the contig polynucleotide sequences disclosed in Table IB.
  • Table IB.l The fifth column in Table IB.l, "ORF (From-To)", provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence "SEQ ED NO:X” that delineate the preferred open reading frame (ORF) shown in the sequence listing and referenced in Table IB.l, column 6, as SEQ ED NO:Y. Where the nucleotide position number 'To" is lower than the nucleotide position number "From", the preferred ORF is the reverse complement of the referenced polynucleotide sequence.
  • the sixth column in Table IB.l provides the corresponding SEQ ED NO:Y for the polypeptide sequence encoded by the preferred ORF delineated in column 5.
  • the invention provides an amino acid sequence comprising, or alternatively consisting of, a polypeptide encoded by the portion of SEQ ED NO:X delineated by "ORF (From-To)". Also provided are polynucleotides encoding such amino acid sequences and the complementary strand thereto.
  • Column 7 in Table IB.l lists residues comprising epitopes contained in the polypeptides encoded by the preferred ORF (SEQ ED NO:Y), as predicted using the algorithm of Jameson and Wolf, (1988) Comp. Appl. Biosci. 4:181-186.
  • polypeptides of the invention comprise, or alternatively consist of, at least one, two, three, four, five or more of the predicted epitopes as described in Table IB. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly.
  • Chromosomal location was determined by finding exact matches to EST and cDNA sequences contained in the NCBI (National Center for Biotechnology Information) UniGene database. Each sequence in the UniGene database is assigned to a "cluster"; all of the ESTs, cDNAs, and STSs in a cluster are believed to be derived from a single gene. Chromosomal mapping data is often available for one or more sequence(s) in a UniGene cluster; this data (if consistent) is then applied to the cluster as a whole. Thus, it is possible to infer the chromosomal location of a new polynucleotide sequence by determining its identity with a mapped UniGene cluster.
  • a modified version of the computer program BLASTN (Altshul, et al., I. Mol. Biol. 215:403-410 (1990), and Gish, and States, Nat. Genet. 3:266-272) (1993) was used to search the UniGene database for EST or cDNA sequences that contain exact or near-exact matches to a polynucleotide sequence of the invention (the 'Query').
  • a sequence from the UniGene database (the 'Subject') was said to be an exact match if it contained a segment of 50 nucleotides in length such that 48 of those nucleotides were in the same order as found in the Query sequence.
  • Table 1B.2 provides an expression profile and library codexount for each of the contig sequences (SEQ ED NO:X) disclosed in Table IB, which can routinely be combined with the information provided in Table 4 and used to determine the tissues, cells, and/or cell line libraries which predominantly express the polynucleotides of the invention.
  • the second number in column 5 represents the number of times a sequence corresponding to the reference polynucleotide sequence was identified in the corresponding tissue/cell source.
  • tissue/cell source identifier codes in which the first two letters are "AR" designate information generated using DNA array technology.
  • cDNAs were amplified by PCR and then transferred, in duplicate, onto the array. Gene expression was assayed through hybridization of first strand cDNA probes to the DNA array.
  • cDNA probes were generated from total RNA extracted from a variety of different tissues and cell lines. Probe synthesis was performed in the presence of 33 P dCTP, using oligo (dT) to prime reverse transcription. After hybridization, high stringency washing conditions were employed to remove non-specific hybrids from the array. The remaining signal, emanating from each gene target, was measured using a Phosphorimager.
  • Phosphor Stimulating Luminescence which reflects the level of phosphor signal generated from the probe hybridized to each of the gene targets represented on the array.
  • a local background signal subtraction was performed before the total signal generated from each array was used to normalize gene expression between the different hybridizations.
  • the value presented after "[array code]:” represents the mean of the duplicate values, following background subtraction and probe normalization.
  • One of skill in the art could routinely use this information to identify normal and/or diseased tissue(s) which show a predominant expression pattern of the corresponding polynucleotide of the invention or to identify polynucleotides which show predominant and/or specific tissue and/or cell expression.
  • Table IC summarizes additional polynucleotides encompassed by the invention (including cDNA clones related to the sequences (Clone ED:), contig sequences (contig identifier (Contig ED:) contig nucleotide sequence identifiers (SEQ ED NO:X)), and genomic sequences (SEQ ED NO:B).
  • the first column provides a unique clone identifier, "Clone ED:”, for a cDNA clone related to each contig sequence.
  • the second column provides the sequence identifier, "SEQ ED NO:X", for each contig sequence.
  • the third column provides a unique contig identifier, "Contig ED:” for each contig sequence.
  • the fourth column provides a BAC identifier "BAC ED NO:A” for the BAC clone referenced in the corresponding row of the table.
  • the fifth column provides the nucleotide sequence identifier, "SEQ ED NO:B" for a fragment of the BAC clone identified in column four of the corresponding row of the table.
  • the sixth column "Exon From- To" provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ ED NO:B which delineate certain polynucleotides of the invention that are also exemplary members of polynucleotide sequences that encode polypeptides of the invention (e.g., polypeptides containing amino acid sequences encoded by the polynucleotide sequences delineated in column six, and fragments and variants thereof).
  • HAUAI83 30 639009 AC010422 783 1-315 2004-2289 2650-2741 3554-3830
  • the polynucleotides or polypeptides, or agonists or antagonists of the present invention can be used in assays to test for one or more biological activities. If these polynucleotides and polypeptides do exhibit activity in a particular assay, it is likely that these molecules may be involved in the diseases associated with the biological activity. Thus, the polynucleotides or polypeptides, or agonists or antagonists could be used to treat the associated disease.
  • the present invention encompasses methods of detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating a disease or disorder.
  • the present invention encompasses a method of treating diabetes mellitus comprising administering to a patient in which such detection, treatment, prevention, and/or amelioration is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to detect, prevent, diagnose, prognosticate, treat, and/or ameliorate diabetes mellitus.
  • the present invention also encompasses methods of detecting, preventing, diagnosing, prognosticating, treating, and/or ameliorating diabetes mellitus; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in Column 3 of Table ID.
  • Table ID provides information related to biological activities for polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof). Table ID also provides information related to assays which may be used to test polynucleotides and polypeptides of the invention (including antibodies, agonists, and/or antagonists thereof) for the corresponding biological activities.
  • the first column (“Gene No.") provides the gene number in the application for each clone identifier.
  • the second column (“cDNA Clone ED:”) provides the unique clone identifier for each clone as previously described and indicated in Table 1A through Table ID.
  • the third column (“AA SEQ ID NO:Y”) indicates the Sequence Listing SEQ ED Number for polypeptide sequences encoded by the corresponding cDNA clones (also as indicated in Tables 1A, Table IB, and Table 2).
  • the fourth column (“Biological Activity”) indicates a biological activity corresponding to the indicated polypeptides (or polynucleotides encoding said polypeptides).
  • the fifth column (“Exemplary Activity Assay”) further describes the corresponding biological activity and also provides information pertaining to the various types of assays which may be performed to test, demonstrate, or quantify the corresponding biological activity.
  • Fluorometric microvolume assay technology is a fluorescence-based system which provides a means to perform nonradioactive cell- and bead- based assays to detect activation of cell signal transduction pathways. This technology was designed specifically for ligand binding and immunological assays. Using this technology, fluorescent cells or beads at the bottom of the well are detected as localized areas of concentrated fluorescence using a data processing system. Unbound flurophore comprising the background signal is ignored, allowing for a wide variety of homogeneous assays.
  • FMAT technology may be used for peptide ligand binding assays, immunofluorescence, apoptosis, cytotoxicity, and bead- based immunocapture assays. See, Miraglia S et. al., "Homogeneous cell and bead based assays for highthroughput screening using flourometric microvolume assay technology," Journal of Biomolecular Screening; 4:193-204 (1999).
  • FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides (including polypeptide fragments and variants) to activate signal transduction pathways.
  • FMAT technology may be used to test, confirm, and/or identify the ability of polypeptides to upregulate production of immunomodulatory proteins (such as, for example, interleukins, GM-CSF, Rantes, and Tumor Necrosis factors, as well as other cellular regulators (e.g. insulin)).
  • immunomodulatory proteins such as, for example, interleukins, GM-CSF, Rantes, and Tumor Necrosis factors, as well as other cellular regulators (e.g. insulin)).
  • Table ID also describes the use of kinase assays for testing, demonstrating, or quantifying biological activity.
  • the phosphorylation and de-phosphorylation of specific amino acid residues e.g. Tyrosine, Serine, Threonine
  • cell-signal transduction proteins provides a fast, reversible means for activation and de-activation of cellular signal transduction pathways.
  • cell signal transduction via phosphorylation/de-phosphorylation is crucial to the regulation of a wide variety of cellular processes (e.g. proliferation, differentiation, migration, apoptosis, etc.).
  • kinase assays provide a powerful tool useful for testing, confirming, and/or identifying polypeptides (including polypeptide fragments and variants) that mediate cell signal transduction events via protein phosphorylation. See e.g., Forrer, P., Tamaskovic R., and Jaussi, R. "Enzyme-Linked Immunosorbent Assay for Measurement of JNK, ERK, and p38 Kinase Activities" Biol. Chem. 379(8-9): 1101-1110 (1998).
  • Calcium Flux assess the ability of polypeptides of the invention (including antibodies and agonists or antagonists of t in pancreatic invention) to mobilize calcium.
  • the FLPR assay may be used to measure influx of calciu beta cells.
  • Cells normally have very low concentrations of cytosolic calcium compared to much higher extracellul calcium. Extracellular factors can cause an influx of calcium, leading to activation of calcium responsi signaling pathways and alterations in cell functions.
  • Exemplary assays that may be used or routinely modified to measure calcium flux by polypeptides of the invention include assays disclosed in: Satin LS, et al, Endocrinology, 136(10):4589- 601 (1995);Mogami H, et al, Endocrinology, 136(7):2960-6 (1995); Richardson SB, et al, Biochem J, ( Pt 3):847-51 (1992); and, Meats, JE, et al, Cell Calcium 1989 Nov-Dec;10(8):535-41 (1989), the contents of each of which is herein incorporated by reference in its entirety.
  • Pancreatic cells that may b used according to these assays are publicly available (e.g., through the ATCC) and/or may be routinely generated.
  • Exemplary pancreatic cells that may be used according to these assays include HITT15 Cell HJTT15 are an adherent epithelial cell line established from Syrian hamster islet cells transformed with SV40. These cells express glucagon, somatostatin, and glucocorticoid receptors. The cells secrete insulin
  • Exemplary assays that may be used or routinely modified to assess the ability of polypeptides and antibodies of the invention (including agonists or antagonists of the invention) to modulate EL-10 production and/or T-cell proliferation include, for example, assays such as disclosed and/or cited in: Robinson, DS, et al, "Th-2 cytokines in allergic disease” Br Med Bull; 56 (4): 956-968 (2000), and Co et al, "T-helper type 2 cell-directed therapy for asthma” Pharmacology & Therapeutics; 88: 187-196 (2000); the contents of each of which are herein incorporated by reference in their entirety.
  • Exemplary cells that may be used according to these assays include Th2 cells.
  • Th2 cells are a class of T cells that secrete EL4, ILIO, EL EL5 and EL6.
  • Factors that induce differentiation and activation of Th2 cells play a major role in the initiation and pathogenesis of allergy and asthma.
  • Primary T helper 2 cells are generated via in vitro culture under Th2 polarizing conditions using peripheral blood lymphocytes isolated from cord blood.
  • Polynucleotides encoding polypeptides of the present invention can be used in assays to test for one or more biological activities.
  • One such biological activity which may be tested includes the ability of polynucleotides and polypeptides of the invention to stimulate upregulation or down-regulation of expression of particular genes and proteins.
  • polynucleotides and polypeptides of the present invention exhibit activity in altering particular gene and protein expression patterns, it is likely that these polynucleotides and polypeptides of the present invention may be involved in, or capable of effecting changes in, diseases associated with the altered gene and protein expression profiles.
  • polynucleotides, polypeptides, or antibodies of the present invention could be used to treat said associated diseases.
  • TaqMan® assays may be performed to assess the ability of polynucleotides (and polypeptides they encode) to alter the expression pattern of particular "target" genes.
  • TaqMan® reactions are performed to evaluate the ability of a test agent to induce or repress expression of specific genes in different cell types.
  • TaqMan® gene expression quantification assays ('TaqMan® assays") are well known to, and routinely performed by, those of ordinary skill in the art.
  • TaqMan® assays are performed in a two step reverse transcription / polymerase chain reaction (RT-PCR). In the first (RT) step, cDNA is reverse transcribed from total RNA samples using random hexamer primers. In the second (PCR) step, PCR products are synthesized from the cDNA using gene specific primers.
  • RT-PCR reverse transcription / polymerase chain reaction
  • the Taqman® PCR reaction exploits the 5' nuclease activity of AmpliTaq Gold ® DNA Polymerase to cleave a Taqman® probe (distinct from the primers) during PCR.
  • the Taqman® probe contains a reporter dye at the 5 '-end of the probe and a quencher dye at the 3' end of the probe. When the probe is intact, the proximity of the reporter dye to the quencher dye results in suppression of the reporter fluorescence.
  • the probe specifically anneals between the forward and reverse primer sites.
  • AmpliTaq Fold DNA Polymerase then cleaves the probe between the reporter and quencher when the probe hybridizes to the target, resulting in increased fluorescence of the reporter (see Figure 2). Accumulation of PCR products is detected directly by monitoring the increase in fluorescence of the reporter dye.
  • probe fragments are displaced from the target, polymerization of the strand continues.
  • the 3 '-end of the probe is blocked to prevent extension of the probe during PCR. This process occurs in every cycle and does not interfere with the exponential accumulation of product.
  • the increase in fluorescence signal is detected only if the target sequence is complementary to the probe and is amplified during PCR. Because of these requirements, any nonspecific amplification is not detected.
  • vector controls or constructs containing the coding sequence for the gene of interest are transfected into cells, such as for example 293T cells, and supernatants collected after 48 hours.
  • multiple primary human cells or human cell lines are used; such cells may include but are not limited to, Normal Human Dermal Fibroblasts, Aortic Smooth Muscle, Human Umbilical Vein Endothelial Cells, HepG2, Daudi, Jurkat, U937, Caco, and THP-1 cell lines.
  • Cells are plated in growth media and growth is arrested by culturing without media change for 3 days, or by switching cells to low serum media and incubating overnight. Cells are treated for 1, 6, or 24 hours with either vector control supernatant or sample supernatant (or purified/partially purified protein preparations in buffer).
  • Total RNA is isolated; for example, by using Trizol extraction or by using the Ambion RNAqueous(TM)-4PCR RNA isolation system. Expression levels of multiple genes are analyzed using TAQMAN, and expression in the test sample is compared to control vector samples to identify genes induced or repressed.
  • Table IE indicates particular disease classes and preferred indications for which polynucleotides, polypeptides, or antibodies of the present invention may be used in detecting, diagnosing, preventing, treating and/or ameliorating said diseases and disorders based on "target" gene expression patterns which may be up- or down-regulated by polynucleotides (and the encoded polypeptides) corresponding to each indicated cDNA Clone ED (shown in Table IE, Column 2).
  • the present invention encompasses a method of detecting, diagnosing, preventing, treating, and/or ameliorating a disease or disorder listed in the "Disease Class" and/or "Preferred Indication” columns of Table IE; comprising administering to a patient in which such detection, diagnosis, prevention, or treatment is desired a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof) in an amount effective to detect, diagnose, prevent, treat, or ameliorate the disease or disorder.
  • the first and second columns of Table ID show the "Gene No.” and "cDNA Clone ID No.”, respectively, indicating certain nucleic acids and proteins (or antibodies against the same) of the invention (including polynucleotide, polypeptide, and antibody fragments or variants thereof) that may be used in detecting, diagnosing, preventing, treating, or ameliorating the disease(s) or disorder(s) indicated in the corresponding row in the "Disease Class” or "Preferred Indication” Columns of Table IE.
  • the present invention also encompasses methods of detecting, diagnosing, preventing, treating, or ameliorating a disease or disorder listed in the "Disease Class” or "Preferred Indication” Columns of Table IE; comprising administering to a patient combinations of the proteins, nucleic acids, or antibodies of the invention (or fragments or variants thereof), sharing similar indications as shown in the corresponding rows in the "Disease Class” or “Preferred Indication” Columns of Table IE.
  • the "Disease Class" Column of Table IE provides a categorized descriptive heading for diseases, disorders, and/or conditions (more fully described below) that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).
  • the "Preferred Indication" Column of Table IE describes diseases, disorders, and/or conditions that may be detected, diagnosed, prevented, treated, or ameliorated by a protein, nucleic acid, or antibody of the invention (or fragment or variant thereof).
  • Cell Line and “Exemplary Targets” Columns of Table IE indicate particular cell lines and target genes, respectively, which may show altered gene expression patterns (i.e., up- or down-regulation of the indicated target gene) in Taqman assays, performed as described above, utilizing polynucleotides of the cDNA Clone ED shown in the corresponding row. Alteration of expression patterns of the indicated “Exemplary Target” genes is correlated with a particular "Disease Class” and/or "Preferred Indication” as shown in the corresponding row under the respective column headings.
  • Cancer in the "Disease Class” Column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof) may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate neoplastic diseases and/or disorders (e.g., leukemias, cancers, etc., as described below under “Hyperproliferative Disorders”).
  • neoplastic diseases and/or disorders e.g., leukemias, cancers, etc., as described below under “Hyperproliferative Disorders”
  • the recitation of “Immune” in the "Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, prevent, treat, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under "Hyperproliferative Disorders"), blood disorders (e.g., as described below under "Immune Activity” "Cardiovascular Disorders” and/or “Blood-Related Disorders”), and infections (e.g., as described below under "Infectious Disease”).
  • neoplastic diseases e.g., as described below under "Hyperproliferative Disorders”
  • blood disorders e.g., as described below under “Immune Activity” "Cardiovascular Disorders” and/or “Blood-Related Disorders”
  • infections e.g., as described below under "
  • Angiogenesis in the "Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diseases and/or disorders relating to neoplastic diseases (e.g., as described below under "Hyperproliferative Disorders"), diseases and/or disorders of the cardiovascular system (e.g., as described below under "Cardiovascular Disorders"), diseases and or disorders involving cellular and genetic abnormalities (e.g., as described below under "Diseases at the Cellular Level"), diseases and/or disorders involving angiogenesis (e.g., as described below under "Anti- Angiogenesis Activity”), to promote or inhibit cell or tissue regeneration (e.g., as described below under “Regeneration”), or to promote wound healing (e.g., as described below under "Wound Healing and Epithelial Cell Proliferation”).
  • neoplastic diseases e
  • Diabetes in the "Disease Class” column indicates that the corresponding nucleic acid and protein, or antibody against the same, of the invention (or fragment or variant thereof), may be used for example, to detect, diagnose, treat, prevent, and/or ameliorate diabetes (including diabetes mellitus types I and II), as well as diseases and/or disorders associated with, or consequential to, diabetes (e.g. as described below under “Endocrine Disorders,” “Renal Disorders,” and “Gastrointestinal Disorders”).
  • Table 2 further characterizes certain encoded polypeptides of the invention, by providing the results of comparisons to protein and protein family databases.
  • the first column provides a unique clone identifier, "Clone ID NO:”, corresponding to a cDNA clone disclosed in Table 1A and/or Table IB.
  • the second column provides the unique contig identifier, "Contig ED:” which allows correlation with the information in Table IB.
  • the third column provides the sequence identifier, "SEQ ED NO:”, for the contig polynucleotide sequences.
  • the fourth column provides the analysis method by which the homology/identity disclosed in the Table was determined.
  • the fifth column provides a description of the PFAM/NR hit identified by each analysis.
  • NR non-redundant protein database
  • PFAM protein families
  • each of the polynucleotides shown in Table IB, column 3 was used to search against the NR database.
  • the computer program BLASTX was used to compare a 6-frame translation of the Query sequence to the NR database (for information about the BLASTX algorithm please see Altshul et al., J. Mol. Biol. 215:403-410 (1990), and Gish and States, Nat. Genet. 3:266-272 (1993).
  • a description of the sequence that is most similar to the Query sequence (the highest scoring 'Subject') is shown in column five of Table 2 and the database accession number for that sequence is provided in column six.
  • the highest scoring 'Subject' is reported in Table 2 if (a) the estimated probability that the match occurred by chance alone is less than 1.0e-07, and (b) the match was not to a known repetitive element.
  • BLASTX returns alignments of short polypeptide segments of the Query and Subject sequences which share a high degree of similarity; these segments are known as High-Scoring Segment Pairs or HSPs.
  • Table 2 reports the degree of similarity between the Query and the Subject for each HSP as a percent identity in Column 7. The percent identity is determined by dividing the number of exact matches between the two aligned sequences in the HSP, dividing by the number of Query amino acids in the HSP and multiplying by 100.
  • the polynucleotides of SEQ ED NO:X which encode the polypeptide sequence that generates an HSP are delineated by columns 8 and 9 of Table 2.
  • the PFAM database PFAM version 2.1, (Sonnhammer, Nucl. Acids Res., 26:320- 322, 1998))consists of a series of multiple sequence alignments; one alignment for each protein family. Each multiple sequence alignment is converted into a probability model called a Hidden Markov Model, or HMM, that represents the position-specific variation among the sequences that make up the multiple sequence alignment (see, e.g., Durbin, et al., Biological sequence analysis: probabilistic models of proteins and nucleic acids, Cambridge University Press, 1998 for the theory of HMMs).
  • HMM Hidden Markov Model
  • HMMER version 1.8 (Sean Eddy, Washington University in Saint Louis) was used to compare the predicted protein sequence for each Query sequence (SEQ ED NO:Y in Table IB) to each of the HMMs derived from PFAM version 2.1.
  • a HMM derived from PFAM version 2.1 was said to be a significant match to a polypeptide of the invention if the score returned by HMMER 1.8 was greater than 0.8 times the HMMER 1.8 score obtained with the most distantly related known member of that protein family.
  • the description of the PFAM family which shares a significant match with a polypeptide of the invention is listed in column 5 of Table 2, and the database accession number of the PFAM hit is provided in column 6.
  • Column 7 provides the score returned by HMMER version 1.8 for the alignment.
  • Columns 8 and 9 delineate the polynucleotides of SEQ ED NO:X which encode the polypeptide sequence which show a significant match to a PFAM protein family. As mentioned, columns 8 and 9 in Table 2, "NT From” and “NT To”, delineate the polynucleotides of "SEQ ID NO:X” that encode a polypeptide having a significant match to the PFAM/NR database as disclosed in the fifth column.
  • the invention provides a protein comprising, or alternatively consisting of, a polypeptide encoded by the polynucleotides of SEQ ED NO:X delineated in columns 8 and 9 of Table 2. Also provided are polynucleotides encoding such proteins, and the complementary strand thereto.
  • nucleotide sequence SEQ ID NO:X and the translated SEQ ED NO:Y are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below.
  • the nucleotide sequences of SEQ ID NO:X are useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ED NO:X or the cDNA contained in ATCC Deposit No:Z. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling immediate applications in chromosome mapping, linkage analysis, tissue identification and/or typing, and a variety of forensic and diagnostic methods of the invention.
  • polypeptides identified from SEQ ED NO:Y may be used to generate antibodies which bind specifically to these polypeptides, or fragments thereof, and/or to the polypeptides encoded by the cDNA clones identified in, for example, Table 1A and/or IB.
  • DNA sequences generated by sequencing reactions can contain sequencing errors.
  • the errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence.
  • the erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence.
  • the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).
  • the present invention provides not only the generated nucleotide sequence identified as SEQ ID NO:X, and a predicted translated amino acid sequence identified as SEQ ED NO:Y, but also a sample of plasmid DNA containing cDNA ATCC Deposit No:Z (e.g., as set forth in columns 2 and 3 of Table 1 A and/or as set forth, for example, in Table IB, 6, and 7).
  • the nucleotide sequence of each deposited clone can readily be determined by sequencing the deposited clone in accordance with known methods. Further, techniques known in the art can be used to verify the nucleotide sequences of SEQ ED NO:X.
  • amino acid sequence of the protein encoded by a particular clone can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence.
  • Partial cDNA clones can be made full-length by utilizing the rapid amplification of cDNA ends (RACE) procedure described in Frohman, M.A., et al., Proc. Nat'l. Acad. Sci. USA, 85:8998-9002 (1988).
  • RACE rapid amplification of cDNA ends
  • RNA Poly A+ or total RNA is reverse transcribed with Superscript II (Gibco/BRL) and an antisense or complementary primer specific to the cDNA sequence.
  • the primer is removed from the reaction with a Microcon Concentrator (Amicon).
  • the first-strand cDNA is then tailed with dATP and terminal deoxynucleotide ttansferase (Gibco/BRL).
  • dATP terminal deoxynucleotide ttansferase
  • the second strand is synthesized from the dA-tail in PCR buffer, Taq DNA polymerase (Perkin-Elmer Cetus), an oligo-dT primer containing three adjacent restriction sites (Xhol, Sail and Clal) at the 5' end and a primer containing just these restriction sites.
  • This double- stranded cDNA is PCR amplified for 40 cycles with the same primers as well as a nested cDNA- specific antisense primer.
  • the PCR products are size-separated on an ethidium bromide-agarose gel and the region of gel containing cDNA products the predicted size of missing protein-coding DNA is removed.
  • cDNA is purified from the agarose with the Magic PCR Prep kit (Promega), restriction digested with Xhol or Sail, and ligated to a plasmid such as pBluescript SKII (Stratagene) at Xhol and EcoRV sites.
  • This DNA is transformed into bacteria and the plasmid clones sequenced to identify the correct protein-coding inserts. Correct 5' ends are confirmed by comparing this sequence with the putatively identified homologue and overlap with the partial cDNA clone. Similar methods known in the art and/or commercial kits are used to amplify and recover 3' ends. Several quality-controlled kits are commercially available for purchase.
  • kit form Similar reagents and methods to those above are supplied in kit form from Gibco/BRL for both 5' and 3' RACE for recovery of full length genes.
  • a second kit is available from Clontech which is a modification of a related technique, SLIC (single-stranded ligation to single-stranded cDNA), developed by Dumas et al., Nucleic Acids Res., 19:5227-32 (1991).
  • SLIC single-stranded ligation to single-stranded cDNA
  • the major differences in procedure are that the RNA is alkaline hydrolyzed after reverse transcription and RNA ligase is used to join a restriction site-containing anchor primer to the first-strand cDNA. This obviates the necessity for the dA-tailing reaction which results in a polyT stretch that is difficult to sequence past.
  • An alternative to generating 5' or 3' cDNA from RNA is to use cDNA library double- sttanded DNA.
  • An asymmetric PCR-amplified antisense cDNA strand is synthesized with an antisense cDNA-specific primer and a plasmid-anchored primer. These primers are removed and a symmetric PCR reaction is performed with a nested cDNA-specific antisense primer and the plasmid-anchored primer.
  • RNA Ligase Protocol For Generating The 5' or 3' End Sequences To Obtain Full Length Genes
  • several methods are available for the identification of the 5' or 3' portions of the gene which may not be present in the original cDNA plasmid. These methods include, but are not limited to, filter probing, clone enrichment using specific probes and protocols similar and identical to 5' and 3' RACE. While the full length gene may be present in the library and can be identified by probing, a useful method for generating the 5' or 3' end is to use the existing sequence information from the original cDNA to generate the missing information. A method similar to 5' RACE is available for generating the missing 5' end of a desired full-length gene.
  • RNA oligonucleotide is ligated to the 5' ends of a population of RNA presumably containing full-length gene RNA transcript and a primer set containing a primer specific to the ligated RNA oligonucleotide and a primer specific to a known sequence of the gene of interest, is used to PCR amplify the 5' portion of the desired full length gene which may then be sequenced and used to generate the full length gene.
  • This method starts with total RNA isolated from the desired source, poly A RNA may be used but is not a prerequisite for this procedure.
  • RNA preparation may then be treated with phosphatase if necessary to eliminate 5' phosphate groups on degraded or damaged RNA which may interfere with the later RNA ligase step.
  • the phosphatase if used is then inactivated and the RNA is treated with tobacco acid pyrophosphatase in order to remove the cap structure present at the 5' ends of messenger RNAs.
  • This reaction leaves a 5' phosphate group at the 5' end of the cap cleaved RNA which can then be ligated to an RNA oligonucleotide using T4 RNA ligase.
  • This modified RNA preparation can then be used as a template for first sttand cDNA synthesis using a gene specific oligonucleotide.
  • the first strand synthesis reaction can then be used as a template for PCR amplification of the desired 5' end using a primer specific to the ligated RNA oligonucleotide and a primer specific to the known sequence of the gene of interest.
  • the resultant product is then sequenced and analyzed to confirm that the 5' end sequence belongs to the relevant gene.
  • the present invention also relates to vectors or plasmids which include such DNA sequences, as well as the use of the DNA sequences.
  • the material deposited with the ATCC (e.g., as described in columns 2 and 3 of Table 1 A, and/or as set forth in Table IB, Table 6, or Table 7) is a mixture of cDNA clones derived from a variety of human tissue and cloned in either a plasmid vector or a phage vector, as described, for example, in Table 1A and Table 7. These deposits are referred to as "the deposits" herein.
  • the tissues from which some of the clones were derived are listed in Table 7, and the vector in which the corresponding cDNA is contained is also indicated in Table 7.
  • the deposited material includes cDNA clones corresponding to SEQ ED NO:X described, for example, in Table 1A and/or Table IB (ATCC Deposit No:Z).
  • a clone which is isolatable from the ATCC Deposits by use of a sequence listed as SEQ ID NO:X may include the entire coding region of a human gene or in other cases such clone may include a substantial portion of the coding region of a human gene.
  • sequence listing may in some instances list only a portion of the DNA sequence in a clone included in the ATCC Deposits, it is well within the ability of one skilled in the art to sequence the DNA included in a clone contained in the ATCC Deposits by use of a sequence (or portion thereof) described in, for example Tables 1A and/or Table IB or Table 2, by procedures hereinafter further described, and others apparent to those skilled in the art.
  • Table 1A and Table 7 is also provided in Table 1A and Table 7 . Each vector is routinely used in the art. The following additional information is provided for convenience.
  • phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene.
  • Vectors pSportl, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0 were obtained from Life Technologies, Inc., P. O. Box 6009, Gaithersburg, MD 20897. All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., et al., Focus 15:59- (1993). Vector lafmid BA (Bento Soares, Columbia University, New York, NY) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue.
  • Vector pCR ® 2.1 which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, CA 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DH10B, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 76:9677-9686 (1988) and Mead, D. et al, Bio/Technology 9: (1991).
  • the present invention also relates to the genes corresponding to SEQ ED NO:X, SEQ ED NO:Y, and/or the deposited clone (ATCC Deposit No:Z).
  • the corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material. Also provided in the present invention are alielic variants, orthologs, and/or species homologs.
  • Procedures known in the art can be used to obtain full-length genes, alielic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of genes corresponding to SEQ ID NO:X or the complement thereof, polypeptides encoded by genes corresponding to SEQ ED NO:X or the complement thereof, and/or the cDNA contained in ATCC Deposit No:Z, using information from the sequences disclosed herein or the clones deposited with the ATCC.
  • alielic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for alielic variants and/or the desired homologue.
  • polypeptides of the invention can be prepared in any suitable manner.
  • Such polypeptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.
  • polypeptides may be in the form of the secreted protein, including the mature form, or may be a part of a larger protein, such as a fusion protein (see below). It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid in purification, such as multiple histidine residues, or an additional sequence for stability during recombinant production.
  • polypeptides of the present invention are preferably provided in an isolated form, and preferably are substantially purified.
  • a recombinantly produced version of a polypeptide, including the secreted polypeptide can be substantially purified using techniques described herein or otherwise known in the art, such as, for example, by the one-step method described in Smith and Johnson, Gene 67:31-40 (1988).
  • Polypeptides of the invention also can be purified from natural, synthetic or recombinant sources using techniques described herein or otherwise known in the art, such as, for example, antibodies of the invention raised against the polypeptides of the present invention in methods which are well known in the art.
  • the present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ ED NO:X, and/or the cDNA sequence contained in ATCC Deposit No:Z.
  • the present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ ED NO:Y, a polypeptide encoded by SEQ ED NO:X or a complement thereof, a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or the polypeptide sequence encoded by a nucleotide sequence in SEQ ED NO:B as defined in column 6 of Table IC.
  • Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ ED NO:Y, a polypeptide encoded by SEQ ED NO:X, a polypeptide encoded by the cDNA contained in ATCC Deposit No:Z, and/or a polypeptide sequence encoded by a nucleotide sequence in SEQ ED NO:B as defined in column 6 of Table IC are also encompassed by the invention.
  • the present invention further encompasses a polynucleotide comprising, or alternatively consisting of, the complement of the nucleic acid sequence of SEQ ED NO:X, a nucleic acid sequence encoding a polypeptide encoded by the complement of the nucleic acid sequence of SEQ ID NO:X, and/or the cDNA contained in ATCC Deposit No:Z.
  • representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in Table IC column 6, or any combination thereof.
  • polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in Table IC column 6, or any combination thereof.
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table IC, column 6, and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ED NO:B (see Table IC, column 5).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table IC, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ED NO:A (see Table IC, column 4).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table IC, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (see Table IC, column 4).
  • Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.
  • representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table IC which correspond to the same Clone ID (see Table IC, column 1), or any combination thereof.
  • Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in column 6 of Table IC which correspond to the same Clone ED (see Table IC, column 1), or any combination thereof.
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table IC which correspond to the same Clone ED (see Table IC, column 1) and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ED NO:B (see Table IC, column 5).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table IC which correspond to the same Clone ED (see Table IC, column 1) and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ED NO:A (see Table IC, column 4).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table IC which correspond to the same Clone ID (see Table IC, column 1) and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ED NO:A (see Table IC, column 4).
  • Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.
  • representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table IC which correspond to the same contig sequence identifier SEQ ID NO:X (see Table IC, column 2), or any combination thereof.
  • Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in column 6 of Table IC which correspond to the same contig sequence identifier SEQ ED NO:X (see Table IC, column 2), or any combination thereof.
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table IC which correspond to the same contig sequence identifier SEQ ED NO:X (see Table IC, column 2) and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ED NO:B (see Table IC, column 5).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table IC which correspond to the same contig sequence identifier SEQ ID NO:X (see Table IC, column 2) and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ED NO:A (see Table IC, column 4).
  • the above- described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table IC which correspond to the same contig sequence identifier SEQ ID NO:X (see Table IC, column 2) and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ED NO:A (See Table IC, column 4).
  • Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.
  • representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in the same row of Table IC column 6, or any combination thereof.
  • Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in the same row of Table IC column 6, or any combination thereof.
  • the polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strand(s) of the sequences delineated in the same row of Table IC column 6, wherein sequentially delineated sequences in the table (i.e. corresponding to those exons located closest to each other) are directly contiguous in a 5' to 3' orientation.
  • above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table IC, column 6, and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ ID NO:B (see Table IC, column 5).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table IC, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO: A (see Table IC, column 4).
  • the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table IC, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC ID NO:A (see Table IC, column 4).
  • Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table IC, and the polynucleotide sequence of SEQ ED NO:X (e.g., as defined in Table IC, column 2) or fragments or variants thereof.
  • Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table IC which correspond to the same Clone ID (see Table IC, column 1), and the polynucleotide sequence of SEQ ED NO:X (e.g., as defined in Table 1A, Table IB, or Table IC) or fragments or variants thereof.
  • the delineated sequence(s) and polynucleotide sequence of SEQ ID NO:X correspond to the same Clone ED.
  • polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in the same row of column 6 of Table IC, and the polynucleotide sequence of SEQ ED NO:X (e.g., as defined in Table 1A, Table IB, or Table IC) or fragments or variants thereof.
  • the delineated sequence(s) and polynucleotide sequence of SEQ ID NO:X correspond to the same row of column 6 of Table IC.
  • Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table IC and the 5' 10 polynucleotides of the sequence of SEQ ED NO:X are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table IC and the 5' 10 polynucleotides of a fragment or variant of the sequence of SEQ ED NO:X are directly contiguous Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of the sequence of SEQ ED NO:X and the 5' 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table IC are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X and the 5' 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table IC are directly contiguous.
  • Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides, are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table IC and the 5' 10 polynucleotides of another sequence in column 6 are directly contiguous.
  • Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table IC and the 5' 10 polynucleotides of another sequence in column 6 corresponding to the same Clone ED (see Table IC, column 1) are directly contiguous. Nucleic acids which hybridize to the complement of these 20 lower stringency conditions, are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one sequence in column 6 corresponding to the same contig sequence identifer SEQ ED NO:X (see Table IC, column 2) are directly contiguous.
  • Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
  • polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table IC and the 5' 10 polynucleotides of another sequence in column 6 corresponding to the same row are directly contiguous.
  • the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table IC is directly contiguous with the 5' 10 polynucleotides of the next sequential exon delineated in Table IC, column 6.
  • Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions are also encompassed by the invention.
  • Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
  • Table 3 Many polynucleotide sequences, such as EST sequences, are publicly available and accessible through sequence databases and may have been publicly available prior to conception of the present invention. Preferably, such related polynucleotides are specifically excluded from the scope of the present invention.
  • each contig sequence (SEQ ED NO:X) listed in the fifth column of Table 1A and/or the fourth column of Table IB preferably excluded are one or more polynucleotides comprising a nucleotide sequence described by the general formula of a-b, where a is any integer between 1 and the final nucleotide minus 15 of SEQ ED NO:X, b is an integer of 15 to the final nucleotide of SEQ ED NO:X, where both a and b correspond to the positions of nucleotide residues shown in SEQ ED NO:X, and where b is greater than or equal to a + 14.
  • polynucleotides comprising a nucleotide sequence described by the general formula of a-b, where a and b are integers as defined in columns 4 and 5, respectively, of Table 3.
  • the polynucleotides of the invention do not consist of at least one, two, three, four, five, ten, or more of the specific polynucleotide sequences referenced by the Genbank Accession No. as disclosed in column 6 of Table 3 (including for example, published sequence in connection with a particular BAC clone).
  • preferably excluded from the invention are the specific polynucleotide sequence(s) contained in the clones corresponding to at least one, two, three, four, five, ten, or more of the available material having the accession numbers identified in the sixth column of this Table (including for example, the actual sequence contained in an identified BAC clone). In no way is this listing meant to encompass all of the sequences which may be excluded by the general formula, it is just a representative example. All references available through these accessions are hereby incorporated by reference in their entirety. Table 3
  • AK026480.1 AL122050.1, AF132676.1, AF061836.1, AL137538.1, AK025015.1, AK000323.1,
  • BC004556.1 AL133565.1, AL122121.1, AB049892.1, Y16645.1, AF177336.1, AB060852.1,
  • AK026534.1 AK024588.1, AF106862.1, AB052191.1, AK026583.1, AJ242859.1, BC008717.1, AK025772.1, S77771.1, AL442082.1, X65873.1, AK026593.1, AB055361.1, AL080060.1, AB051158.1, X82434.1, AF097996.1, AK026597.1, AK025414.1, AF183393.1, AL133557.1, AF252872.1, AL133113.1, BC002733.1, AK026462.1, AL080124.1, BC008488.1, BC008070.1, AL133080.1, AB055370.1, AK025339.1, AB055315.1, AL512733.1, AL162006.1, AL049452.1,

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Abstract

La présente invention a trait à des polypeptides humains sécrétés, et des molécules isolées d'acide nucléique codant pour lesdits peptides, utiles pour le diagnostic et le traitement du diabète sucré et/ou des conditions liées au diabète. L'invention concerne également des anticorps de liaison à de tels polypeptides. L'invention concerne en outre des vecteurs, des cellules hôtes, et des procédés de recombinaison et de synthèse pour la production desdits polynucléotides, polypeptides, et/ou anticorps. L'invention concerne aussi des procédés de criblage permettant l'identification d'agonistes et d'antagonistes des polynucléotides et polypeptides de l'invention. Enfin, l'invention concerne des procédés et des compositions permettant l'inhibition ou l'amélioration de la production et de la fonction desdits polypeptides.
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Families Citing this family (80)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7196164B2 (en) 1997-07-08 2007-03-27 Human Genome Sciences, Inc. Secreted protein HHTLF25
JP2002513295A (ja) 1997-07-08 2002-05-08 ヒューマン・ジェノム・サイエンシズ・インコーポレイテッド １２３種類のヒト分泌タンパク質
US20060052321A1 (en) 2002-04-05 2006-03-09 Raitano Arthur B Nucleic acid and corresponding protein entitled 98P4B6 useful in treatment and detection of cancer
US20040141975A1 (en) 1998-06-01 2004-07-22 Raitano Arthur B. Nucleic acid and corresponding protein entitled 98P4B6 useful in treatment and detection of cancer
US20030149531A1 (en) 2000-12-06 2003-08-07 Hubert Rene S. Serpentine transmembrane antigens expressed in human cancers and uses thereof
PT1086223E (pt) 1998-06-01 2009-11-03 Agensys Inc Novos antigénios transmembranares em serpentina expressos em cancros humanos e suas utilizações
US6833438B1 (en) 1999-06-01 2004-12-21 Agensys, Inc. Serpentine transmembrane antigens expressed in human cancers and uses thereof
WO2000042073A1 (fr) 1999-01-15 2000-07-20 Biogen, Inc. Antagonistes de la proteine tweak et du recepteur de tweak, et leur utilisation pour traiter des affections immunitaires
EP1666490A3 (fr) * 2000-07-25 2006-11-02 Genentech, Inc. Polypeptides sécrétés et transmembranaires ainsi que les acides nucléiques codant pour ceux-ci
US7291719B2 (en) 2000-07-25 2007-11-06 Genentech, Inc. PRO4332 antibodies
EP1364021A2 (fr) 2000-10-31 2003-11-26 Diadexus, Inc. Compositions et methodes associees a des genes et a des proteines specifiques du colon
US20020111302A1 (en) * 2000-11-30 2002-08-15 Y. Tom Tang Novel nucleic acids and polypeptides
US7736654B2 (en) * 2001-04-10 2010-06-15 Agensys, Inc. Nucleic acids and corresponding proteins useful in the detection and treatment of various cancers
US20030119112A1 (en) 2001-06-20 2003-06-26 Genentech, Inc. Compositions and methods for the diagnosis and treatment of disorders involving angiogenesis
US7494646B2 (en) 2001-09-06 2009-02-24 Agensys, Inc. Antibodies and molecules derived therefrom that bind to STEAP-1 proteins
ES2532757T3 (es) 2001-09-06 2015-03-31 Agensys, Inc. Ácido nucleico y proteína correspondiente denominados STEAP-1 útiles en el tratamiento y la detección de cáncer
WO2003033727A1 (fr) 2001-10-12 2003-04-24 Yamanouchi Pharmaceutical Co., Ltd. Procede de criblage d'un inhibiteur de la mort cellulaire
CA2887716C (fr) 2002-04-09 2017-01-24 Biogen Idec Ma Inc. Procede pour traiter les etats lies a tweak
AU2003900747A0 (en) * 2003-02-18 2003-03-06 Garvan Institute Of Medical Research Diagnosis and treatment of pancreatic cancer
US7709610B2 (en) 2003-05-08 2010-05-04 Facet Biotech Corporation Therapeutic use of anti-CS1 antibodies
US20050025763A1 (en) 2003-05-08 2005-02-03 Protein Design Laboratories, Inc. Therapeutic use of anti-CS1 antibodies
WO2005019258A2 (fr) * 2003-08-11 2005-03-03 Genentech, Inc. Compositions et methodes de traitement de maladies relatives au systeme immunitaire
EP1670947B1 (fr) 2003-09-23 2015-04-01 University Of North Carolina At Chapel Hill Technique et composition de correlation de polymorphisme nucleotidique simple dans le gene vitamine k epoxyde reductase et le dosage de warfarine
JP4777251B2 (ja) * 2003-10-14 2011-09-21 バクスター・インターナショナル・インコーポレイテッド ビタミンｋエポキシド還元酵素複合体サブユニット１ｖｋｏｒｃ１、クマリンおよびその誘導体の治療標的
EP1714154A2 (fr) * 2004-02-03 2006-10-25 Bayer HealthCare AG Agents diagnostiques et therapeutiques pour des maladies associes a la glutamate carboxypeptidase (pgcp) plasmatique
US20050186577A1 (en) 2004-02-20 2005-08-25 Yixin Wang Breast cancer prognostics
AU2004319915B9 (en) 2004-04-22 2011-12-22 Agensys, Inc. Antibodies and molecules derived therefrom that bind to STEAP-1 proteins
NZ553581A (en) 2004-09-01 2011-04-29 Dynavax Tech Corp Methods and compositions for inhibition of innate immune responses and autoimmunity using immunoregulatory polynucleotides with a TGC sequence
WO2006026808A1 (fr) * 2004-09-07 2006-03-16 Telethon Institute For Child Health Research Methode de diagnostic et/ou de prediction du developpement d'une affection allergique
JP2008512099A (ja) * 2004-09-07 2008-04-24 テレソン インスティテュート フォー チャイルド ヘルス リサーチ アレルギー性疾患の処置または予防のための物質
AU2006214179A1 (en) 2005-02-17 2006-08-24 Biogen Idec Ma Inc. Treating neurological disorders
CN101124320A (zh) 2005-02-28 2008-02-13 巴克斯特国际公司 重组共表达维生素k环氧化物还原酶亚基1以提高依赖维生素k的蛋白质表达
EP1861499A4 (fr) 2005-03-15 2008-09-03 Univ North Carolina Procédés et compositions pour la production de protéines actives dépendantes de la vitamine k
EP1885388B1 (fr) 2005-05-10 2013-09-11 Biogen Idec MA Inc. Traitement et evaluation des troubles inflammatoires
WO2006138219A2 (fr) 2005-06-13 2006-12-28 Biogen Idec Ma Inc. Procedes d'evaluation de patients
US20090054307A1 (en) * 2005-09-01 2009-02-26 Howard Florey Institute Of Experimental Physiology And Medicine Prophylactic and therapeutic agents and uses therefor
GB0521488D0 (en) * 2005-10-21 2005-11-30 Ares Trading Sa Integral membrane protein
US7572618B2 (en) 2006-06-30 2009-08-11 Bristol-Myers Squibb Company Polynucleotides encoding novel PCSK9 variants
ES2636089T3 (es) 2006-10-27 2017-10-05 Genentech, Inc. Anticuerpos e inmunoconjugados y usos para los mismos
JP5588175B2 (ja) 2006-11-07 2014-09-10 メルク・シャープ・アンド・ドーム・コーポレーション Ｐｃｓｋ９のアンタゴニスト
US8093222B2 (en) 2006-11-27 2012-01-10 Isis Pharmaceuticals, Inc. Methods for treating hypercholesterolemia
CA2680832A1 (fr) * 2007-03-27 2008-10-02 Merck & Co., Inc. Procede de detection de pcsk9 secretee, autogeneree
JOP20080381B1 (ar) 2007-08-23 2023-03-28 Amgen Inc بروتينات مرتبطة بمولدات مضادات تتفاعل مع بروبروتين كونفيرتاز سيتيليزين ككسين من النوع 9 (pcsk9)
JP5749492B2 (ja) 2007-10-26 2015-07-15 ダイナバックス テクノロジーズ コーポレイション 免疫応答と自己免疫を阻害する方法及び組成物
CA2707182C (fr) * 2007-11-30 2017-12-12 Siemens Healthcare Diagnostics Inc. Fragments de recepteur de l'adiponectine et leurs procedes d'utilisation
AR070316A1 (es) 2008-02-07 2010-03-31 Merck & Co Inc Antagonistas de pcsk9 (proproteina subtilisina-kexina tipo 9)
AR070315A1 (es) 2008-02-07 2010-03-31 Merck & Co Inc Anticuerpos 1b20 antagonistas de pcsk9
WO2010068526A1 (fr) 2008-12-12 2010-06-17 Merck Sharp & Dohme Corp. Immunodosage de pcsk9
AU2010313381A1 (en) 2009-10-30 2012-04-12 Merck Sharp & Dohme Corp. AX1 and AX189 PCSK9 antagonists and variants
CN102639150A (zh) 2009-10-30 2012-08-15 默沙东公司 Ax213和ax132 pcsk9拮抗剂和变体
BR112012032240A2 (pt) 2010-06-16 2019-09-24 Dynavax Technologies Corporation método de tratamento utilizando inibidores tlr7 e/ou tlr9
CN105061534A (zh) 2010-09-22 2015-11-18 艾丽奥斯生物制药有限公司 取代的核苷酸类似物
WO2012088222A2 (fr) 2010-12-21 2012-06-28 The University Of North Carolina At Chapel Hill Procédés et compositions pour la production de protéines actives dépendantes de la vitamine k
AU2012358804B2 (en) 2011-12-22 2018-04-19 Alios Biopharma, Inc. Substituted phosphorothioate nucleotide analogs
US8916538B2 (en) 2012-03-21 2014-12-23 Vertex Pharmaceuticals Incorporated Solid forms of a thiophosphoramidate nucleotide prodrug
NZ630805A (en) 2012-03-22 2016-01-29 Alios Biopharma Inc Pharmaceutical combinations comprising a thionucleotide analog
US8945560B1 (en) 2014-07-15 2015-02-03 Kymab Limited Method of treating rheumatoid arthritis using antibody to IL6R
US9045548B1 (en) 2014-07-15 2015-06-02 Kymab Limited Precision Medicine by targeting rare human PCSK9 variants for cholesterol treatment
US8986691B1 (en) 2014-07-15 2015-03-24 Kymab Limited Method of treating atopic dermatitis or asthma using antibody to IL4RA
US9017678B1 (en) 2014-07-15 2015-04-28 Kymab Limited Method of treating rheumatoid arthritis using antibody to IL6R
US8883157B1 (en) 2013-12-17 2014-11-11 Kymab Limited Targeting rare human PCSK9 variants for cholesterol treatment
US9051378B1 (en) 2014-07-15 2015-06-09 Kymab Limited Targeting rare human PCSK9 variants for cholesterol treatment
US8992927B1 (en) 2014-07-15 2015-03-31 Kymab Limited Targeting human NAV1.7 variants for treatment of pain
US9023359B1 (en) 2014-07-15 2015-05-05 Kymab Limited Targeting rare human PCSK9 variants for cholesterol treatment
US9067998B1 (en) 2014-07-15 2015-06-30 Kymab Limited Targeting PD-1 variants for treatment of cancer
US9045545B1 (en) 2014-07-15 2015-06-02 Kymab Limited Precision medicine by targeting PD-L1 variants for treatment of cancer
US9034332B1 (en) 2014-07-15 2015-05-19 Kymab Limited Precision medicine by targeting rare human PCSK9 variants for cholesterol treatment
US9914769B2 (en) 2014-07-15 2018-03-13 Kymab Limited Precision medicine for cholesterol treatment
US8986694B1 (en) 2014-07-15 2015-03-24 Kymab Limited Targeting human nav1.7 variants for treatment of pain
US8980273B1 (en) 2014-07-15 2015-03-17 Kymab Limited Method of treating atopic dermatitis or asthma using antibody to IL4RA
US9682218B2 (en) 2013-12-23 2017-06-20 Carefusion 2200, Inc. Pleurodesis device and method
GB201403775D0 (en) 2014-03-04 2014-04-16 Kymab Ltd Antibodies, uses & methods
US9139648B1 (en) 2014-07-15 2015-09-22 Kymab Limited Precision medicine by targeting human NAV1.9 variants for treatment of pain
US9150660B1 (en) 2014-07-15 2015-10-06 Kymab Limited Precision Medicine by targeting human NAV1.8 variants for treatment of pain
WO2018083248A1 (fr) 2016-11-03 2018-05-11 Kymab Limited Anticorps, combinaisons comprenant des anticorps, biomarqueurs, utilisations et procédés
AU2017382323B2 (en) 2016-12-23 2024-06-13 President And Fellows Of Harvard College Gene editing of PCSK9
JP7247101B2 (ja) 2017-04-03 2023-03-28 エフ・ホフマン－ラ・ロシュ・アクチェンゲゼルシャフト Ｓｔｅａｐ－１に結合する抗体
EP3642383B1 (fr) 2017-06-22 2022-12-21 The Procter & Gamble Company Films comprenant une couche hydrosoluble et un revêtement inorganique déposé en phase vapeur
EP3641951B1 (fr) 2017-06-22 2023-09-20 The Procter & Gamble Company Films comprenant une couche hydrosoluble et un revêtement organique déposé en phase vapeur
WO2021207711A2 (fr) 2020-04-09 2021-10-14 Verve Therapeutics, Inc. Arn guides modifiés chimiquement pour l'édition du génome avec cas9

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000004140A1 (fr) * 1998-07-15 2000-01-27 Human Genome Sciences, Inc. 71 proteines humaines secretees
WO2002026931A2 (fr) * 2000-09-25 2002-04-04 Human Genome Sciences, Inc. 71 proteines humaines secretees

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100236393B1 (ko) * 1996-02-02 1999-12-15 나까니시 히로유끼 사람성장호르몬을 함유하는 의약제제
US5858716A (en) * 1997-05-30 1999-01-12 Smithkline Beecham Corporation H2CAA71 polynucleotides

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000004140A1 (fr) * 1998-07-15 2000-01-27 Human Genome Sciences, Inc. 71 proteines humaines secretees
WO2002026931A2 (fr) * 2000-09-25 2002-04-04 Human Genome Sciences, Inc. 71 proteines humaines secretees

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
HSU SHEAU YU ET AL: "Characterization of two LGR genes homologous to gonadotropin and thyrotropin receptors with extracellular leucine-rich repeats and a G protein-coupled, seven-transmembrane region" MOLECULAR ENDOCRINOLOGY, vol. 12, no. 12, December 1998 (1998-12), pages 1830-1845, XP002528150 ISSN: 0888-8809 *
LOH E D ET AL: "Molecular Characterization of a Novel Glycoprotein Hormone G-Protein-Coupled Receptor" BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, ACADEMIC PRESS INC. ORLANDO, FL, US, vol. 282, no. 3, 6 April 2001 (2001-04-06), pages 757-764, XP003002810 ISSN: 0006-291X *
See also references of WO03004622A2 *

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