Compositions and methods for the treatment and diagnosis of cardiovascular disease using rchd534 as a target

Abstract

The present invention relates to methods and compositions for the treatment and diagnosis of cardiovascular disease, including, but not limited to, atherosclerosis, ischemia/reperfusion, hypertension, restenosis, and arterial inflammation. Specifically, the present invention identifies and describes genes which are differentially expressed in cardiovascular disease states, relative to their expression in normal, or non-cardiovascular disease states, and/or in response to manipulations relevant to cardiovascular disease. Further, the present invention identifies and describes genes via the ability of their gene products to interact with gene products involved in cardiovascular disease. Still further, the present invention provides methods for the identification and therapeutic use of compounds as treatments of cardiovascular disease. Moreover, the present invention provides methods for the diagnostic monitoring of patients undergoing clinical evaluation for the treatment of cardiovascular disease, and for monitoring the efficacy of compounds in clinical trials. Additionally, the present invention describes methods for the diagnostic evaluation and prognosis of various cardiovascular diseases, and for the identification of subjects exhibiting a predisposition to such conditions.

What is claimed is:  
     
         1 . An isolated nucleic acid containing the following nucleotide sequence: 
 rchd005 (SEQ ID NO.:1),    rchd024 (SEQ ID NO.:2),    rchd032 (SEQ ID NO.:3),    rchd036 (SEQ ID NO.:4),    rchd502 (SEQ ID NO.:5),    rchd523 (SEQ ID NO.:6),    rchd528 (SEQ ID NO.:7), or    rchd534 (SEQ ID NO.:36).    or the nucleotide sequence of a gene or gene fragment contained in the following clone as deposited with the NRRL:    pRCHD005 (in NRRL Accession No. B-21376),    pRCHD024 (in NRRL Accession No. B-21377),    pRCHD032 (in NRRL Accession No. B-21378),    pRCHD036 (in NRRL Accession No. B-21379),    PRCHD502 (in NRRL Accession No. B-21380),    PRCHD523 (in NRRL Accession No. B-21381),    pFCHD523 (in NRRL Accession No. ),    PRCHD528 (in NRRL Accession No. B-21382), or    pFCHD534 (in NRRL Accession No. ).    
     
     
         2 . An isolated nucleic acid which hybridizes under stringent conditions to the nucleotide sequence of  claim 1  or its complement, or to the gene or gene fragment contained in the clone of  claim 1  as deposited with the NRRL.  
     
     
         3 . An isolated nucleic acid which encodes an amino acid sequence encoded by the nucleotide sequence of  claim 1  or its complement, or the gene or gene fragment contained in the clone of  claim 1  as deposited with the NRRL.  
     
     
         4 . A nucleotide vector containing the nucleotide sequence of  claim 1 ,  2  or  3 .  
     
     
         5 . An expression vector containing the nucleotide sequence of  claim 1 ,  2  or  3  in operative association with a nucleotide regulatory element that controls expression of the nucleotide sequence in a host cell.  
     
     
         6 . A genetically engineered host cell containing the nucleotide sequence of  claim 1 ,  2  or  3 .  
     
     
         7 . A genetically engineered host cell containing the nucleotide sequence of  claim 1 ,  2  or  3  in operative association with a nucleotide regulatory element that controls expression of the nucleotide sequence in the host cell.  
     
     
         8 . A substantially pure gene product encoded by the nucleic acid of  claim 1 ,  2 , or  3 .  
     
     
         9 . An antibody that immunospecifically binds the gene product of  claim 8 .  
     
     
         10 . A transgenic animal in which the nucleic acid of  claim 1 ,  2  or  3  is an expressed transgene contained in the genome of the animal.  
     
     
         11 . A transgenic animal in which expression of genomic sequences encoding the gene product of  claim 8  is prevented or suppressed.  
     
     
         12 . A method for diagnosing cardiovascular disease, comprising detecting, in a patient sample, a gene or its gene product which is differentially expressed in cardiovascular disease states.  
     
     
         13 . The method of  claim 12  in which the cardiovascular disease is atherosclerosis.  
     
     
         14 . The method of  claim 12  in which the cardiovascular disease is ischemia/reperfusion.  
     
     
         15 . The method of  claim 12  in which the cardiovascular disease is hypertension.  
     
     
         16 . The method of  claim 12  in which the cardiovascular disease is restenosis.  
     
     
         17 . The method of  claim 12  in which the gene is up-regulated in individuals genetically predisposed to cardiovascular disease.  
     
     
         18 . The method of  claim 17  in which the gene encodes a Na—K—Cl cotransporter protein homologue, an rchd024 protein, and rchd032 protein, an rchd036 protein, a homolog of rat matrin F/G protein, an endoperoxide synthase type II protein, an rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         19 . The method of  claim 12  in which the gene is down-regulated in individuals genetically predisposed to cardiovascular disease.  
     
     
         20 . The method of  claim 19  in which the gene encodes a glutathione peroxidase protein or a Bcl-2 protein.  
     
     
         21 . The method of  claim 12  in which the gene is up-regulated by treatment with IL-1.  
     
     
         22 . The method of  claim 21  in which the gene encodes an Na—K—Cl cotransporter protein homologue, an rchd024 protein, an rchd032 protein, an rchd036 protein, or an endoperoxide synthase type II protein.  
     
     
         23 . The method of  claim 12  in which the gene is up-regulated by treatment with shear stress.  
     
     
         24 . The method of  claim 23  in which the gene encodes an Na—K—Cl cotransporter protein homologue, an rchd 0 24 protein, a rat matrin F/G protein homologue, an endoperoxide synthase type II protein, an rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         25 . The method of  claim 12  wherein the gene is down-regulated by treatment of individuals with a high fat/high cholesterol diet.  
     
     
         26 . The method of  claim 25  in which the gene encodes a glutathione peroxidase protein or a Bcl-2 protein.  
     
     
         27 . A method for treating cardiovascular disease, comprising administering a compound that modulates the synthesis or expression of a target gene, or the activity of a target gene product to a patient in need of such treatment.  
     
     
         28 . The method of  claim 27  in which the cardiovascular disease is atherosclerosis.  
     
     
         29 . The method of  claim 27  in which the cardiovascular disease is ischemia/reperfusion.  
     
     
         30 . The method of  claim 27  in which the cardiovascular disease is hypertension.  
     
     
         31 . The method of  claim 27  in which the cardiovascular disease is restenosis.  
     
     
         32 . The method of  claim 27  in which the compound inhibits the expression of the target gene, or the synthesis or activity of the target gene product.  
     
     
         33 . The method of  claim 32  in which the gene encodes a Na—K—Cl cotransporter protein homologue, an rchd024 protein, and rchd032 protein, an rchd036 protein, a homolog of rat matrin F/G protein, an endoperoxide synthase type II protein, an chd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         34 . The method of  claim 27  in which the compound is an ntisense or ribozyme molecule that blocks translation of the arget gene.  
     
     
         35 . The method of  claim 34  in which the gene encodes a Na—K—Cl cotransporter protein homologue, an rchd024 protein, and rchd032 protein, an rchd036 protein, a homologue of rat atrin F/G protein, an endoperoxide synthase type II protein, and rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         36 . The method of  claim 27  in which the compound is complementary to the 5′ region of the target gene and blocks transcription via triple helix formation.  
     
     
         37 . The method of  claim 36  in which the gene encodes a Na—K—Cl cotransporter protein homologue, an rchd024 protein, and rchd032 protein, an rchd036 protein, a homologue of rat matrin F/G protein, an endoperoxide synthase type II protein, and rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         38 . The method of  claim 27  in which the compound is an antibody that neutralizes the activity of the target gene product.  
     
     
         39 . The method of  claim 38  in which the gene product is a Na—K—Cl cotransporter protein homologue, an rchd024 protein, and rchd032 protein, an rchd036 protein, a homologue of rat matrin F/G protein, an endoperoxide synthase type II protein, and rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         40 . The method of  claim 27  in which the compound enhances the expression of the target gene, or the synthesis or activity the target gene product.  
     
     
         41 . The method of  claim 40  in which the target gene encodes Bcl-2 or glutathione peroxidase.  
     
     
         42 . A method for treating cardiovascular disease, comprising administering nucleic acid encoding an active target gene product to a patient in need of such treatment.  
     
     
         43 . The method of  claim 42  in which the nucleic acid encodes Bcl-2 or glutathione peroxidase.  
     
     
         44 . A method for treating cardiovascular disease, comprising administering an effective amount of a target gene product to a patient in need of such therapy.  
     
     
         45 . The method of  claim 44  in which the gene product is Bcl-2 or glutathione peroxidase.  
     
     
         46 . A method of monitoring the efficacy of a compound in clinical trials for the treatment of cardiovascular disease, comprising detecting, in a patient sample, a gene or its gene product which is differentially expressed in cardiovascular disease states.  
     
     
         47 . The method of  claim 46  in which the cardiovascular disease is atherosclerosis.  
     
     
         48 . The method of  claim 46  in which the cardiovascular disease is ischemia/reperfusion.  
     
     
         49 . The method of  claim 46  in which the cardiovascular disease is hypertension.  
     
     
         50 . The method of  claim 46  in which the cardiovascular disease is restenosis.  
     
     
         51 . The method of  claim 46  in which the gene is up-regulated in individuals genetically predisposed to cardiovascular disease.  
     
     
         52 . The method of  claim 51  in which the gene encodes a Na—K—Cl cotransporter protein homologue, an rchd024 protein, and rchd032 protein, an rchd036 protein, a homolog of rat matrin F/G protein, an endoperoxide synthase type II protein, and rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         53 . The method of  claim 46  in which the gene is down-regulated in individuals genetically predisposed to cardiovascular disease.  
     
     
         54 . The method of  claim 53  in which the gene encodes a glutathione peroxidase protein or a Bcl-2 protein.  
     
     
         55 . The method of  claim 46  in which the gene is up-regulated by treatment with IL-1.  
     
     
         56 . The method of  claim 55  in which the gene encodes an Na—K—Cl cotransporter protein homologue, an rchd024 protein, an rchd032 protein, an rchd036 protein, or an endoperoxide synthase type II protein.  
     
     
         57 . The method of  claim 46  in which the gene is up-regulated by treatment with shear stress.  
     
     
         58 . The method of  claim 57  in which the gene encodes an Na—K—Cl cotransporter protein homologue, an rchd024 protein, a rat matrin F/G protein homologue, an endoperoxide synthase type II protein, an rchd523 protein, an rchd528 protein, or an rchd534 protein.  
     
     
         59 . The method of  claim 46  wherein the gene is down-regulated by treatment of individuals with a high fat/high cholesterol diet.  
     
     
         60 . The method of  claim 59  in which the gene encodes a glutathione peroxidase protein or a Bcl-2 protein.  
     
     
         61 . A method for identifying a compound that modulates the activity of a multiple transmembrane domain receptor target gene product, comprising: 
 contacting a first cell expressing the multiple transmembrane domain receptor target gene product wtith a test compound and an activator of the multiple transmembrane domain receptor target gene product, measuring the level of intracellular calcium release within the first cell and comparing the level to that of a second multiple transmembrane domain receptor target gene product-expressing cell which has been contacted with the activator but not with the test compound so that if the level of intracellular calcium release within the first cells differs from that of the second cell, a compound which modulates the activity of a multiple transmembrane domain receptor target gene product has been identified.    
     
     
         62 . The method of  claim 61  wherein the multiple transmembrane domain receptor target gene product is an rchd523 gene product.  
     
     
         63 . The method of  claim 61  wherein the cell is a Xenopus oocyte cell.  
     
     
         64 . The method of  claim 61  wherein the cell is a myeloma cell.  
     
     
         65 . The method of  claim 18  in which the gene encodes an rchd523 protein.  
     
     
         66 . The method of  claim 18  in which the gene encodes an rchd534 protein.