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 treating a subject having heart failure, comprising: directly administering into the lumen of the coronary artery of the subject, an effective amount of an adeno-associated virus (AAV) vector comprising a nucleic acid encoding a constitutively active fragment of human phosphatase inhibitor-1 protein sequence set forth in SEQ ID NO: 2 truncated at the C-terminus at amino acid 65 comprising an aspartic acid at position 35 (T35D), that inhibits human type-1 phosphatases (PP1), operably linked to a promoter capable of directing expression in heart cells, thereby expressing the constitutively active fragment of human phosphatase inhibitor-1 protein in the heart of the subject to increase β-adrenergic responsiveness.
2. The method of claim 1 , wherein the promoter is a constitutive promoter.
3. The method of claim 1 , wherein the promoter is expressed in a cardiac muscle.
4. The method of claim 1 , wherein the promoter is selected from the group consisting of a Cytomegalovirus (CMV) promoter, a cardiac specific cardiac troponin T promoter, a myosin heavy chain promoter and a myosin light chain promoter.
5. The method of claim 1 , wherein the AAV vector is selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 and AAV9.
6. The method of claim 1 , wherein the AAV vector is administered in an amount effective to result in myocyte shortening, to lower the time constant for relaxation, tau (T), and/or to accelerate calcium signal decay in the heart of the subject.
7. The method of claim 1 , wherein the AAV vector is administered in an amount effective to increase in phosphorylation of serine 16 of phospholamban in the heart of the subject.
8. The method of claim 1 , wherein the AAV vector is administered in an amount effective to improve the end-systolic pressure dimension relationship in the heart of the subject.
9. The method of claim 1 , wherein the heart failure is associated with a reduced β-adrenergic response in the heart of the subject.
10. The method of claim 1 , wherein the heart failure comprises ischemia, arrhythmia, myocardial infarction, abnormal heart contractility, or abnormal Ca 2+ metabolism.
11. The method of claim 1 , wherein the heart failure is congestive heart failure.
12. The method of claim 1 , wherein the AAV vector is administered in an amount effective to increase cardiac contractility and reduce morphological deterioration associated with cardiac remodeling in the heart of the subject.
13. The method of claim 12 , wherein reduced 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 reduction is present if expression of the 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.
14. The method of claim 1 , wherein the amount of the AAV vector is between 1×10 11 and 1×10 16 infectious units.
15. The method of claim 5 , wherein the AAV vector is AAV6.