Recombinant adeno-associated virus capsids with enhanced human skeletal muscle tropism
The present invention relates to variant AAV capsid polypeptides, wherein the variant AAV capsid polypeptides exhibit increased transduction and/or tropism in human muscle tissue or cells as compared non-variant parent capsid polypeptides.
1. A variant adeno-associated virus (AAV) capsid polypeptide, wherein said variant AAV capsid polypeptide exhibits increased transduction or tropism in human skeletal muscle tissue or cells as compared to a non-variant parent capsid polypeptide, wherein said variant AAV capsid polypeptide comprises a sequence selected from the group consisting of AAV-NP6 (SEQ ID NO: 1), AAV-NP20 (SEQ ID NO: 2), AAV-NP22(SEQ ID NO: 3), AAV-NP36 (SEQ ID NO: 4), AAV-NP66 (SEQ ID NO: 5), AAV-NP81 (SEQ ID NO: 6), and AAV-NP94 (SEQ ID NO: 7).
2. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide exhibits increased transduction as compared to a non-variant parent capsid polypeptide.
3. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide exhibits increased tropism as compared to a non-variant parent capsid polypeptide.
4. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide further exhibits an enhanced neutralization profile as compared to a non-variant parent capsid polypeptide.
5. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide further exhibits increased transduction or tropism in one or more non-muscle human tissues as compared to a non-variant parent capsid polypeptide.
6. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide exhibits increased transduction of human muscle tissue or cells in vivo as compared to a non-variant parent capsid polypeptide.
7. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide exhibits increased transduction of human muscle tissue or cells in vitro as compared to a non-variant parent capsid polypeptide.
8. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide exhibits increased transduction of a human muscle tissue explant ex vivo as compared to a non-variant parent capsid polypeptide.
9. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide is part of a functional AAV capsid, wherein said functional AAV capsid packages a nucleic acid sequence selected from the group consisting of a non-coding RNA, a protein coding sequence, an expression cassette, a multi-expression cassette, a sequence for homologous recombination, a genomic gene targeting cassette, and a therapeutic expression cassette.
10. The variant AAV capsid polypeptide of claim 9 , wherein said nucleic acid sequence is contained within an AAV vector.
11. The variant AAV capsid polypeptide of claim 9 , wherein said expression cassette is a CRISPR/CAS expression system.
12. The variant AAV capsid polypeptide of claim 9 , wherein said therapeutic expression cassette encodes a therapeutic protein or antibody.
13. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP22 (SEQ ID NO: 3).
14. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP66 (SEQ ID NO: 5).
15. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP6 (SEQ ID NO: 1).
16. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP20 (SEQ ID NO: 2).
17. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP36 (SEQ ID NO: 4).
18. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP81 (SEQ ID NO: 6).
19. The variant AAV capsid polypeptide of claim 1 , wherein said variant AAV capsid polypeptide sequence is AAV-NP94 (SEQ ID NO: 7).
20. The variant AAV capsid polypeptide of claim 9 , wherein said variant AAV capsid polypeptide results in increased nucleic acid expression as compared to a non-variant parent capsid polypeptide.
21. A variant adeno-associated virus (AAV) capsid polypeptide, wherein said variant AAV capsid polypeptide exhibits enhanced neutralization profile as compared to a non-variant parent capsid polypeptide, wherein said variant AAV capsid polypeptide comprises a sequence selected from the group consisting of AAV-NP6 (SEQ ID NO: 1), AAV-NP20 (SEQ ID NO: 2), AAV-NP22 (SEQ ID NO: 3), AAV-NP36 (SEQ ID NO: 4), AAV-NP66 (SEQ ID NO: 5), AAV-NP81 (SEQ ID NO: 6), and AAV-NP94 (SEQ ID NO: 7).
22. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide exhibits an enhanced neutralization profile against pooled human immunoglobulins as compared to a non-variant parent capsid polypeptide.
23. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide further exhibits increased transduction or tropism in human skeletal muscle tissue or cells as compared to a non-variant parent capsid polypeptide.
24. The variant AAV capsid polypeptide of claim 23 , wherein said variant AAV capsid polypeptide exhibits increased transduction as compared to a non-variant parent capsid polypeptide.
25. The variant AAV capsid polypeptide of claim 23 , wherein said variant AAV capsid polypeptide exhibits increased tropism as compared to a non-variant parent capsid polypeptide.
26. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide further exhibits increased transduction or tropism in one or more non-muscle human tissues as compared to a non-variant parent capsid polypeptide.
27. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide exhibits increased transduction of human muscle tissue or cells in vivo as compared to a non-variant parent capsid polypeptide.
28. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide exhibits increased transduction of human muscle tissue or cells in vitro as compared to a non-variant parent capsid polypeptide.
29. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide exhibits increased transduction of a human muscle tissue explant ex vivo as compared to a non-variant parent capsid polypeptide.
30. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide is part of a functional AAV capsid, wherein said functional AAV capsid packages a nucleic acid sequence selected from the group consisting of a non-coding RNA, a protein coding sequence, an expression cassette, a multi-expression cassette, a sequence for homologous recombination, a genomic gene targeting cassette, and a therapeutic expression cassette.
31. The variant AAV capsid polypeptide of claim 30 , wherein said nucleic acid sequence is contained within an AAV vector.
32. The variant AAV capsid polypeptide of claim 30 , wherein said expression cassette is a CRISPR/CAS expression system.
33. The variant AAV capsid polypeptide of claim 30 , wherein said therapeutic expression cassette encodes a therapeutic protein or antibody.
34. The variant AAV capsid polypeptide of claim 30 , wherein said variant AAV capsid polypeptide results in increased nucleic acid expression as compared to a non-variant parent capsid polypeptide.
35. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP22 (SEQ ID NO: 3).
36. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP66 (SEQ ID NO: 5).
37. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP6 (SEQ ID NO: 1).
38. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP20 (SEQ ID NO: 2).
39. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP36 (SEQ ID NO: 4).
40. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP81 (SEQ ID NO: 6).
41. The variant AAV capsid polypeptide of claim 21 , wherein said variant AAV capsid polypeptide sequence is AAV-NP94 (SEQ ID NO: 7).