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 T 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 recombinant adeno-associated virus (rAAV) virion, comprising a vector genome comprising an expression cassette comprising a polynucleotide encoding myosin binding protein C (MYBPC3) operatively linked to a promoter, the expression cassette flanked by a 5′ inverted terminal repeat (ITR) and a 3′ ITR; wherein the vector genome comprises at most 4.8 kbp; and wherein the expression cassette shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 95.
2. The rAAV virion of claim 1 , wherein the promoter is a cardiac troponin T promoter.
3. The rAAV virion of claim 2 , therein the cardiac troponin T promoter comprises about 400 hp.
4. The rAAV virion of claim 3 , wherein the cardiac troponin T promoter shares at least 90% identity to the polynucleotide sequence of SEQ 11) NO: 3.
5. The rAAV virion of claim 3 , wherein the cardiac troponin I promoter shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 3.
6. The rAAV virion of claim 3 , wherein the cardiac troponin I promoter shares 100% identity to the polynucleotide sequence of SEQ ID NO: 3.
7. The rAAV virion of claim 1 , wherein MYBPC3 shares at least 95% identity to the amino acid sequence of SEQ ID NO: 103.
8. The rAAV virion of claim 1 , wherein the polynucleotide encoding MYBPC3 shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 86.
9. The rAAV virion of claim 1 , wherein the vector genome comprises at most 4.7 kbp.
10. The rAAV virion of claim 1 , wherein the vector genome comprises 4.7 kbp.
11. The rAAV virion 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.
12. The rAAV virion of claim 1 , wherein the expression cassette shares 100% identity to the polynucleotide sequence of SEQ ID NO: 95.
13. The rAAV virion of claim 1 , wherein the vector genome shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 102.
14. The rAAV virion of claim 1 , wherein the vector genome shares 100% identity to the polynucleotide sequence of SEQ ID NO: 102.
15. The rAAV virion of claim 1 , wherein the rAAV virion is a serotype AAV9 virion.
16. A pharmaceutical composition comprising the rAAV virion of claim 1 .
17. The pharmaceutical composition of claim 16 , wherein the vector genome shares at least 95% identity to SEQ ID NO: 102.