Methods and compositions relating to a cardiac-specific nuclear regulatory factor
AssigneeBoard of Regents, The University of Texas System
InventorsEric N. Olson, Da-Zhi Wang
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Abstract
The present invention relates to a novel cardiac-specific transcription factor, myocardin. This molecule modulates the development and differentiation of cardiomyocytes and is a potent inhibitor of cell growth. Methods to exploit these observations are provided and include respecifiying non-cardiac cells into cardiac cells, stimulating cardiac tissue regeneration, and methods for treating cardiomyopathies, myocardial infarction.
Claims126
An isolated polynucleotide encoding a myocardin polypeptide.
An isolated nucleic acid segment comprising at least 15 contiguous nucleotides of SEQ ID NO:1, SEQ ID NO:25, SEQ ID NO:27 or SEQ ID NO:29.
An expression cassette comprising a polynucleotide encoding a myocardin polypeptide operably linked to a regulatory sequence.
A transformed host cell comprising a polynucleotide encoding a myocardin polypeptide and a promoter heterologous to the polypeptide coding region, wherein said promoter directs expression of said myocardin polypeptide.
A method of using a host cell comprising an expression cassette comprising a polynucleotide encoding a myocardin polypeptide and a promoter active in said host cell comprising culturing the host cell under conditions suitable for the expression of the myocardin polypeptide.
A peptide of 8 to about 50 residues comprising at least 8 consecutive residues of SEQ ID NO:2, SEQ ID NO:26, SEQ ID NO:28 or SEQ ID NO:30.
A fusion protein comprising a myocardin protein or peptide fused to a second protein or peptide.
A method of modulating the phenotype of a non-cardiomyocyte cell to include one or more phenotypic functions of a cardiomyocyte cell comprising introducing into said non-cardiac cell an expression cassette comprising a polynucleotide encoding a myocardin polypeptide and a promoter active in said non-cardiac cell, wherein said promoter directs the expression of said polypeptide.
A method of generating a cardiomyocyte comprising introducing into a cardiac fibroblast an expression vector comprising a polynucleotide encoding a myocardin polypeptide and a promoter active in said fibroblast, wherein said promoter directs the expression of said polypeptide.
A method of stimulating cardiac tissue regeneration comprising inhibiting the function of myocardin in a post-mitotic cardiomyocyte.
A method of expressing a myocardin polypeptide in a host cell comprising introducing into said host cells an expression vector comprising a polynucleotide encoding a myocardin polypeptide, said polynucleotide being positioned under control of a promoter operable in said host cell.
A monoclonal antibody that binds immunologically to a polypeptide comprising SEQ ID NO:2, SEQ ID NO:26, SEQ ID NO:28 or SEQ ID NO:30 or an antigenic fragment thereof.
A polyclonal antisera, antibodies of which bind immunologically to a polypeptide comprising SEQ ID NO:2, SEQ ID NO:26, SEQ ID NO:28 or SEQ ID NO:30 or an antigenic fragment thereof.
A hybridoma cell that produces a monoclonal antibody that binds immunologically to a polypeptide comprising SEQ ID NO:2, SEQ ID NO:26, SEQ ID NO:28 or SEQ ID NO:30 or an antigenic fragment thereof.
A non-human transgenic animal comprising an expression cassette, wherein said expression cassette comprises a polynucleotide encoding a myocardin peptide or protein and a promoter operable in eukaryotic cells, said promoter being heterologous to the myocardin peptide or protein encoding region.
A non-human transgenic animal comprising a defective germ-line myocardin allele.
A method of treating a heart disease, including cardiomyopathy comprising administering to an animal suffering therefrom an expression cassette comprising a polynucleotide encoding a myocardin peptide or protein and a promoter operable in eukaryotic cells.
A method of treating a heart disease, including cardiomyopathy comprising the step of providing to an animal suffering therefrom a myocardin antisense nucleic acid.
A method of decreasing mortality in a subject with heart failure comprising inhibiting the function of myocardin in post-mitotic cardiomyocytes in the subject.
A method of decreasing mortality in a subject with heart failure comprising increasing the level of myocardin in fibroblasts to generate cardiomyocytes in the subject.
A method of decreasing morbidity in a subject with heart failure comprising inhibiting the function of myocardin in post-mitotic cardiomyocytes in the subject.
A method of decreasing morbidity in a subject with heart failure comprising increasing the level of myocardin in fibroblasts to generate cardiomyocytes in the subject.
A method of screening for a candidate substance for an effect on myocardin regulation of cardiomyocyte development comprising:
- (a) providing myocardin and GATA to a cell;
- (b) admixing myocardin and GATA in the presence of said candidate substance;
- and
- (c) measuring the effect of said candidate substance on the expression of a cardiac lineage marker,
- wherein a difference in the expression of said cardiac lineage marker, as compared to an untreated cell, indicates that said candidate substance effects myocardin regulation of cardiomyocyte development.
A method of screening for a modulator of myocardin expression comprising:
- (a) providing a cell that expresses a myocardin polypeptide;
- (b) contacting said myocardin polypeptide with a candidate substance;
- and
- (c) measuring the expression of myocardin,
- wherein a difference in myocardin expression, indicates that said candidate substance is a modulator of myocardin expression.
A method of screening a candidate substance for myocardin binding activity comprising:
- (a) providing a myocardin polypeptide;
- (b) contacting the myocardin polypeptide with the candidate substance;
- and
- (c) determining the binding of the candidate substance to the myocardin polypeptide.
Dependencies are inferred from the claim text.