Modulating phosphatase activity in cardiac cells
Expression of a phosphatase inhibitor in heart cells can be used to treat cardiac disorders, e.g., heart failure. Decreasing phosphatase activity can improve β-adrenergic responsiveness.
1. A method of increasing cardiac contractility and reducing morphological deterioration associated with cardiac remodeling in a subject with existing heart failure, said method comprising:
directly introducing in vivo an effective amount of a viral vector into the lumen of a coronary artery of the heart of the subject by percutaneous intracoronary gene transfer, wherein said viral vector is an adeno-associated viral vector (AAV) comprising a nucleic acid sequence encoding a polypeptide consisting of amino acids 1-65 of SEQ ID NO: 2, wherein threonine at position 35 of SEQ ID NO: 2 is replaced with an aspartic acid; and wherein said nucleic acid sequence is operably linked to a promoter capable of directing expression in the heart;
thereby expressing the fragment in the heart of the subject in an amount effective to increase cardiac contractility and reduce morphological deterioration associated with cardiac remodeling in the subject with existing heart failure.
2. The method of claim 1 , wherein the promoter is a tissue specific promoter, a smooth muscle specific promoter, a cardiac specific promoter, or a viral promoter.
3. The method of claim 1 , wherein the promoter is a constitutive promoter.
4. The method of claim 2 , wherein the tissue specific promoter is a cardiac troponin T promoter, myosin heavy chain or the myosin light chain promoter, cardiac myosin promoter, troponin T promoter, BNP promoter, alpha actin promoter, or a SM22a promoter.
5. The method of claim 1 , wherein the viral vector is selected from the group consisting of an adeno-associated virus-1 (AAV1), AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 and AAV9.
6. The method of claim 1 , wherein the increase in contractility is determined by increased myocyte shortening (myocyte length change), rates of myocyte cell shortening (dL/dt) and relengthening (−dL/dt), a lower time constant for relaxation (tau(τ)), and accelerated calcium signal decay.
7. The method of claim 1 , wherein the protection against morphological deterioration associated with cardiac remodeling is determined by measuring the heart-to-body weight ratio, infarct size, and the presence of cardiac fibrosis, wherein protection is present if expression of said human phosphatase inhibitor-1 results in a reduced heart-to-body weight ratio, a decreased infarct size, or reduced cardiac fibrosis as compared to a control.
8. The method of claim 1 , wherein the heart failure comprises ischemia, arrhythmia, myocardial infarction, abnormal heart contractility, or abnormal Ca 2 + metabolism.
9. The method of claim 1 , wherein the coronary artery is the left anterior descending artery (LAD) or the distal circumflex artery (LCX).
10. The method of claim 1 , wherein the method further comprises restricting blood flow to one or more of the great coronary vein (GCV), the middle cardiac vein (MCV), or the anterior interventricular vein (AIV).
11. The method of claim 9 , wherein the vector is introduced into the lumen of the coronary artery after ischemic preconditioning of the left anterior descending artery (LAD) and/or the distal circumflex artery (LCX).
12. The method of claim 1 , wherein the vector is introduced into the lumen of the coronary artery with a catheter.
13. The method of claim 9 , wherein the coronary artery is the distal circumflex artery (LCX).
14. The method of claim 9 , wherein the coronary vessel is the left anterior descending artery (LAD).
15. The method of claim 1 , wherein the amount of the viral vector is between 1×10 11 and 1×10 16 plaque forming units.
16. The method of claim 1 , wherein the viral vector is adeno-associated virus serotype 9 (AAV9).