Method of detecting cardiac hypertrophy through probe hybridization and gene expression analysis

Abstract

The present invention is based upon the identification of genes which are differentially expressed in hypertrophic cardiac tissue as compared to normal cardiac tissue. Accordingly, the present invention provides nucleotide sequences of genes selected from the group consisting of CH-1, CH-2, CH-3, CH-4, CH-5, CH-6, CH-7, CH-8, and CH-9 and amino acid sequences of their encoded proteins, as well as derivatives (e.g., fragments) and analogs thereof. The invention also provides therapeutic methods and pharmaceutical compositions which are based on the promotion or inhibition of the function of the differentially expressed genes. The invention further provides methods of diagnosis, prognosis and screening for a disposition for diseases or disorders associated with cardiac hypertrophy. Methods for screening for modulators of the protein products of the differentially expressed genes in cardiac hypertrophy tissue are additionally provided.

Description

[0001] This application is a divisional of U.S. patent application Ser. No. 09 / 554,169 filed Aug. 21, 2000, now U.S. Pat. No. ______, issued on ______, which is the nationalization of PCT Patent Application No. PCT / US98 / 23953 filed on Nov. 10, 1998, which claims priority to U.S. patent application Ser. No. 09 / 189,618, filed on Nov. 9, 1998, now U.S. Pat. No. 6,610,480, issued on Aug. 26, 2003, which claims priority to U.S. Provisional Application Serial No. 60 / 065,048, filed Nov. 10, 1997.[0002] Heart failure is often defined as the inability of the heart to deliver a supply of oxygenated blood sufficient to meet the metabolic needs of peripheral tissues, both at rest and during exercise. See generally, Hutter, Jr., "Congestive Heart Failure", in Scientific American: Medicine, Volume 1 (1:11), eds. Dale and Federman (Scientific American, Inc. 1994). Heart failure is a common outcome of hypertension or post-myocardial infarction and is a major contributor to cardiovascular morbidity ...

AI Technical Summary

Benefits of technology

[0087] PCR can be carried out, e.g., by use of a Perkin-Ehner Cetus thermal cycler and Taq polymerase (Gene Amp.RTM.). The DNA being amplified can include mRNA or cDNA or genomic DNA from any eukaryotic species. One can choose to synthesize several different degenerate primers, for use in the PCR reactions. It is also possible to vary the stringency of hybridization conditions used in priming the PCR reactions, to amplify nucleic acid homologs (e.g., to obtain CH sequences from species other than humans or to obtain human sequences with homology to CH) by allowing for greater or lesser degrees of nucleotide sequence similarity between the known nucleotide sequence and the nucleic acid homolog being isolated. For cross species hybridization, low stringency conditions are preferred. For same species hybridization, moderately stringent conditions are preferred.

[0102] According to the invention, techniques described for the production of single chain antibodies (U.S. Pat. No. 4,946,778) can be adapted to produce CH-specific single chain antibodies. An additional embodiment of the invention utilizes the techniques described for the construction of Fab expression libraries (Huse et al., Science 246:1275-81 (1989)) to allow rapid and easy identification of monoclonal Fab fragments with the desired specificity for CH or CH derivatives, or analogs thereof. Non-human antibodies can be "humanized" by known methods (See, e.g., U.S. Pat. No. 5,225,539).

[0119] Patients suffering from heart failure may also be predisposed to cardiac hypertrophy. By way of example but not by way of limitation, coronary artery disease, cardiomyopathy, myocarditis, aortic stenosis, hypertension, coarctation of the aorta, aortic regurgitation, mitral regurgitation, left-to-right shunts, restrictive cardiomyopathy, ischemic heart disease, pericardial tamponade, constrictive pericarditis, or restrictive cardiomyopathy can increase the likelihood that a patient will suffer a cardiac hypertrophy.

Problems solved by technology

Heart failure is a common outcome of hypertension or post-myocardial infarction and is a major contributor to cardiovascular morbidity and mortality.

Pressure overload leads to myocardial hypertrophy and the remodeling of muscular and collagenous compartments of the myocardium where the accumulation of fibrillar collagen is known to impair myocardial stiffness.

However, the technical limitations of the candidate gene paradigm, in which specific genes of interest have been tested one or several at a time for their association with heart disease, mean that relatively little information is available compared to the entire complement of genes expressed in the myocardium.

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