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 of 4.7 kilobase pairs (kb), comprising an expression cassette comprising a polynucleotide encoding full-length myosin binding protein C (MYBPC3) operatively linked to a promoter, the expression cassette flanked by a 5′ inverted terminal repeat MR) and a 3′ ITR;
wherein the 4.7 kb rAAV virion is able to treat hypertrophic cardiomyopathy via significantly improved cardiac function based on ejection fraction and significantly decreased hypertrophy compared to a 5.4 kb rAAV virion expressing the MYBPC3 protein.
2. The rAAV virion of claim 1 , wherein the promoter is a cardiac troponin T promoter.
3. The rAAV virion of claim 2 , wherein the cardiac troponin T promoter comprises about 400 bp.
4. The rAAV virion of claim 2 , wherein the cardiac troponin T promoter comprises about 400 bp and wherein the cardiac troponin T promoter shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 3.
5. The rAAV virion of claim 2 , wherein the cardiac troponin T promoter shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 3.
6. The rAAV virion of claim 2 , wherein the cardiac troponin T promoter shares at least 95% identity to the polynucleotide sequence of SEQ ID NO: 3.
7. The rAAV virion of claim 2 , wherein the cardiac troponin T promoter comprises the polynucleotide sequence of SEQ ID NO: 3.
8. The rAAV virion of claim 1 , wherein the MYBPC3 shares at least 90% identity to the amino acid sequence of SEQ ID NO: 103.
9. The rAAV virion of claim 1 , wherein the polynucleotide encoding the MYBPC3 shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 86.
10. 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.
11. The rAAV virion of claim 1 , wherein the expression cassette shares at least 80% identity to the polynucleotide sequence of SEQ ID NO: 95.
12. The rAAV virion of claim 1 , wherein the expression cassette shares at least 90% identity to the polynucleotide sequence of SEQ ID NO: 95.
13. The rAAV virion of claim 1 , wherein the vector genome shares at least 85% identity to the polynucleotide sequence of SEQ. ID NO: 102.
14. The rAAV virion of claim 1 , wherein the vector genome shares at least 90% 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. The rAAV virion of claim 1 , wherein the MYBPC3 comprises the amino acid sequence of SEQ ID NO: 103.
17. The rAAV virion of claim 1 , wherein the polynucleotide encoding the MYBPC3 comprises the polynucleotide sequence of SEQ ID NO: 86.
18. The rAAV virion of claim 1 , wherein the expression cassette comprises the polynucleotide sequence of SEQ ID NO: 95.
19. The rAAV virion of claim 1 , wherein the vector genome comprises the polynucleotide sequence of SEQ ID NO: 102.