Gene therapy vectors for treating heart disease
The present disclosure provides methods and compositions useful for the treatment or prevention of heart disease. In particular, the present disclosure provides a vector comprising a modified troponin promoter operatively linked to a therapeutic gene product for the treatment or prevention of heart disease, e.g., cardiomyopathy. The gene product may be MYBPC3. The disclosure also provides recombinant adeno-associated virus (rAAV) virions, rAAV viral genomes, and expression cassettes and pharmaceutical compositions thereof. The disclosure further provides methods for treating a disease or disorder, such as heart disease.
1. A method of treating a hypertrophic cardiomyopathy or a cardiomyopathy caused by a myosin binding protein C (MYBPC3) mutation in a subject in need thereof, comprising:
administering systemically to the subject or directly to the heart tissue of the subject a recombinant adeno-associated virus (rAAV) virion, comprising a vector genome of at most 4.8 kb,
wherein the vector genome comprises an expression cassette comprising a polynucleotide encoding full-length MYBPC3 operatively linked to a cardiac troponin T promoter, the expression cassette flanked by a 5′ inverted terminal repeat (ITR) and a 3′ ITR.
2. The method of claim 1 , wherein the cardiac troponin T promoter comprises about 400 bp.
3. The method of claim 2 , wherein the cardiac troponin T promoter shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 3.
4. The method of claim 2 , wherein the cardiac troponin T promoter comprises the polynucleotide sequence of SEQ ID NO:3.
5. The method of claim 1 , wherein the MYBPC3 shares at least 90% identity to the amino acid sequence of SEQ ID NO: 103.
6. The method of claim 1 , wherein the polynucleotide encoding the MYBPC3 shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 86.
7. The method of claim 1 , wherein the 5′ ITR comprises a sequence that shares 95% identity to the polynucleotide sequence of SEQ ID NO: 96; and the 3′ ITR comprises a sequence that shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 97.
8. The method of claim 1 , wherein the cardiac troponin T promoter shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 3.
9. The method of claim 1 , wherein the cardiac troponin T promoter shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 3.
10. The method of claim 1 , wherein the expression cassette shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 95.
11. The method of claim 1 , wherein the expression cassette shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 95.
12. The method of claim 1 , wherein the vector genome shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 102.
13. The method of claim 1 , wherein the vector genome shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 102.
14. The method of claim 1 , wherein the rAAV virion is a serotype AAV9 virion.
15. The method of claim 1 , which is a method of treating a cardiomyopathy caused by a MYBPC3 mutation in a subject.
16. The method of claim 1 , which is a method of treating a hypertrophic cardiomyopathy.
17. The method of claim 16 , wherein the hypertrophic cardiomyopathy is caused by a MYBPC3 mutation in the subject.
18. The method of claim 17 , wherein the cardiac troponin T promoter comprises about 400 bp.
19. The method of claim 18 , wherein the cardiac troponin T promoter shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 3.
20. The method of claim 17 , wherein the MYBPC3 shares at least 90% identity to the amino acid sequence of SEQ ID NO: 103.
21. The method of claim 17 , wherein the polynucleotide encoding the MYBPC3 shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 86.
22. The method of claim 17 , wherein the 5′ ITR comprises a sequence that shares 95% identity to the polynucleotide sequence of SEQ ID NO: 96; and the 3′ ITR comprises a sequence that shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 97.
23. The method of claim 17 , wherein the cardiac troponin T promoter shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 3.
24. The method of claim 17 , wherein the cardiac troponin T promoter shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 3.
25. The method of claim 17 , wherein the expression cassette shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 95.
26. The method of claim 17 , wherein the expression cassette shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 95.
27. The method of claim 17 , wherein the vector genome shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 102.
28. The method of claim 17 , wherein the method increases cardiac function assessed by ejection fraction in the subject.
29. The method of claim 1 , wherein the method increases cardiac function assessed by ejection fraction in the subject.
30. The method of claim 1 , wherein the administering systemically is administering intravenously, intra-arterially or intraperitoneally.
31. The method of claim 30 , wherein the administering systemically is administering intravenously.
32. The method of claim 1 , wherein the administering directly to the heart tissue is by direct injection into the heart, cardiac catheterization or intracoronary administration.
33. The method of claim 32 , wherein the administering directly to the heart tissue is by intracardiac catheter delivery via retrograde coronary sinus infusion.
34. The method of claim 1 , wherein the MYBPC3 comprises the amino acid sequence of SEQ ID NO: 103.
35. The method of claim 1 , wherein the polynucleotide encoding the MYBPC3 comprises the polynucleotide sequence of SEQ ID NO:86.
36. The method of claim 1 , wherein the expression cassette comprises the polynucleotide sequence of SEQ ID NO:95.
37. The method of claim 1 , wherein the vector genome comprises the polynucleotide sequence of SEQ ID NO:102.
38. The method of claim 1 , wherein the MYBPC3 mutation is a deletion of the gene encoding MYBPC3.
39. The method of claim 1 , wherein the subject is a human.
40. The method of claim 1 , wherein the cardiac troponin T promoter consists of the polynucleotide sequence of SEQ ID NO:3.