Cell and gene based methods to improve cardiac function

Patent metadata

Field	Value
Publication number	US-2016186139-A1
Application number	US-201214122226-A
Country	US
Kind code	A1
Filing date	May 29, 2012
Priority date	May 26, 2011
Publication date	Jun 30, 2016
Grant date	—

Abstract

Compositions and methods for improving cardiac function, myocardial contractility and relaxation in a mammal are provided. Cardiomyocytes transfected with one or more expression vectors comprising a ribonucleotide reductase subunit R1-encoding nucleic acid sequence and a ribonucleotide reductase subunit R2-encoding nucleic acid sequence operably linked to a promoter are grafted to a mammalian myocardium. Also provided are compositions and methods for delivering dATP to a myocardium through grafting of donor cells overexpressing R1 and R2. dATP is thereby produced in situ and delivered through gap junctions established between donor cells and host cardiomyocytes. Alternatively, viral vector(s) having the R1 and R2-encoding construct(s) are administered to the mammal directly. Improvement of cardiac function can also be effected by administration of vectors comprising a nucleic acid sequence encoding a L48Q, 61 Q, or L57Q cTnC variant.

First claim

1 . A method of improving cardiac function in a mammal comprising:
grafting cardiomyocytes to the myocardium of said mammal, wherein said cardiomyocytes contain a first expression vector comprising a first nucleic acid sequence encoding ribonucleotide reductase subunit R1 and a second expression vector comprising a second nucleic acid sequence encoding ribonucleotide reductase subunit R2, said first nucleic acid sequence and second nucleic acid sequence being operably linked to a promoter that induces overexpression of R1 and R2.

2 - 28 . (canceled)

29 . A method of improving cardiac function in a mammal comprising administering a first viral vector comprising a first nucleic acid sequence encoding ribonucleotide reductase subunit R1 and a second viral vector comprising a second nucleic acid sequence encoding ribonucleotide reductase subunit R2, said first nucleic acid sequence and second nucleic acid sequence being operably linked to a cardiac-specific promoter.

30 . The method of claim 29 , wherein the first and second viral vectors are adeno-associated viral vectors.

31 . The method of claim 29 , wherein the mammal is human.

32 . The method of claim 29 , wherein the first and second viral vectors are administered systemically.

33 . The method of claim 29 , wherein the first and second viral vectors are administered intravenously.

34 . The method of claim 29 , wherein the first and second viral vectors are administered by intra-myocardial injection.

35 . The method of claim 29 , wherein at least one of said first and second viral vectors further comprises a transduction reporter.

36 . The method of claim 29 , wherein the first and second viral vectors are the same vector.

37 - 62 . (canceled)

63 . A pharmaceutical composition comprising a first viral vector comprising a first nucleic acid sequence encoding ribonucleotide reductase subunit R1 and a second viral vector comprising a second nucleic acid sequence encoding ribonucleotide reductase subunit R2, said first nucleic acid sequence and second nucleic acid sequence being operably linked to a cardiac-specific promoter.

64 - 65 . (canceled)

66 . A pharmaceutical composition comprising a viral vector comprising a nucleic acid sequence encoding a cTnC variant having an amino acid substitution selected from L48Q, I61Q, and L57Q.

67 - 68 . (canceled)

69 . A pharmaceutical composition comprising cardiomyocytes containing a first expression vector comprising a first nucleic acid sequence encoding ribonucleotide reductase subunit R1 and a second expression vector comprising a second nucleic acid sequence encoding ribonucleotide reductase subunit R2, said first nucleic acid sequence and second nucleic acid sequence being operably linked to a promoter that induces overexpression of R1 and R2.

70 . (canceled)

71 . A method of improving cardiac function in an individual in need thereof comprising:
administering a viral vector comprising a nucleic acid sequence encoding a cTnC variant having an amino acid substitution selected from L48Q, L57Q, and I61Q, said nucleic acid sequence being operably linked to a cardiac-specific promoter.

72 - 88 . (canceled)

89 . A method of improving cardiac function in a mammal comprising administering a first expression vector comprising a first nucleic acid sequence encoding ribonucleotide reductase subunit R1 and a second expression vector comprising a second nucleic acid sequence encoding ribonucleotide reductase subunit R2, said first nucleic acid sequence and second nucleic acid sequence being operably linked to a cardiac-specific promoter.

90 . A method of improving cardiac function in a mammal comprising administering an expression vector comprising a first nucleic acid sequence encoding ribonucleotide reductase subunit R1 and a second nucleic acid sequence encoding ribonucleotide reductase subunit R2, said first nucleic acid sequence and second nucleic acid sequence being operably linked to a cardiac-specific promoter.

Assignees

• Univ Washington

Inventors

• Regnier Michael
• Laflamme Michael
• Murry Charles
• Korte F Steven
• Lundy Scott
• Hauschka Stephen Denison
• Chamberlain Jeffrey S

Classifications

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• A61K35/34

  Muscles; Smooth muscle cells; Heart; Cardiac stem cells; Myoblasts; Myocytes; Cardiomyocytes (vascular smooth muscle A61K35/44) · CPC title

• C12N15/86

  Viral vectors · CPC title

• A61P9/10

  for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis · CPC title

• C12N9/0093

  acting on CH or CH2 groups (1.17) · CPC title