Compositions and methods for treating non-age-associated hearing impairment in a human subject
Provided herein are compositions that include at least two different nucleic acid vectors, where each of the at least two different vectors includes a coding sequence that encodes a different portion of an otoferlin protein, and the use of these compositions to treat hearing loss in a subject.
1. A plurality of recombinant adeno-associated viral (rAAV) vectors comprising:
a) a first rAAV vector comprising a nucleic acid sequence of SEQ ID NO: 96; and
b) a second rAAV vector comprising a nucleic acid sequence of SEQ ID NO: 105.
2. The plurality of rAAV vectors of claim 1 , wherein the first and second rAAV vectors are each encapsulated by an AAV capsid.
3. The plurality of rAAV vectors of claim 2 , wherein the AAV capsid encapsulating the first rAAV vector is a serotype selected from any one of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, or Anc80.
4. The plurality of rAAV vectors of claim 2 , wherein the AAV capsid encapsulating the second rAAV vector is a serotype selected from any one of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, or Anc80.
5. The plurality of rAAV vectors of claim 2 , wherein the first rAAV vector is encapsulated by an Anc80 capsid and the second rAAV vector is encapsulated by an Anc80 capsid.
6. The plurality of rAAV vectors of claim 5 , wherein the Anc80 capsids comprise the polypeptide sequence of SEQ ID NO: 109.
7. A composition comprising the plurality of rAAV vectors of claim 1 , wherein the composition further comprises one or more pharmaceutically acceptable carriers, diluents, or excipients.
8. The composition of claim 7 , wherein the composition is formulated for intra-cochlear administration.
9. The composition of claim 8 , wherein the composition is formulated to comprise a synthetic perilymph solution.
10. A method of expressing a recombinant full-length otoferlin protein in a mammalian cell, the method comprising administering a plurality of recombinant adeno-associated viral (rAAV) vectors comprising:
a) a first rAAV vector comprising a nucleic acid sequence of SEQ ID NO: 96; and
b) a second rAAV vector comprising a nucleic acid sequence of SEQ ID NO: 105;
to the mammalian cell, wherein the mammalian cell has reduced expression, lack of expression or dysfunction of otoferlin.
11. The method of claim 10 , wherein the mammalian cell is an inner hair cell.
12. The method of claim 10 , wherein the mammalian cell is a human cell.
13. The method of claim 10 , wherein the mammalian cell comprises a defective otoferlin gene.
14. The method of claim 10 , wherein the first and second rAAV vectors are each encapsulated by an AAV capsid.
15. The method of claim 14 , wherein the AAV capsid encapsulating the first rAAV vector is a serotype selected from any one of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, or Anc80.
16. The method of claim 14 , wherein the AAV capsid encapsulating the second rAAV vector is a serotype selected from any one of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, or Anc80.
17. The method of claim 14 , wherein the first rAAV vector is encapsulated by an Anc80 capsid and the second rAAV vector is encapsulated by an Anc80 capsid.
18. The method of claim 16 , wherein the Anc80 capsids comprise the polypeptide sequence of SEQ ID NO: 109.
19. A method of treating hearing loss in a subject having otoferlin related hearing loss, the method comprising administering a plurality of recombinant adeno-associated viral (rAAV) vectors comprising:
a) a first rAAV vector comprising a nucleic acid sequence of SEQ ID NO: 96; and
b) a second rAAV vector comprising a nucleic acid sequence of SEQ ID NO: 105;
into the cochlea of the subject, wherein the subject has reduced expression, lack of expression, or dysfunction of otoferlin.
20. The method of claim 19 , wherein the subject is a human.
21. The method of claim 19 , wherein the first and second rAAV vectors are each encapsulated by an AAV capsid.
22. The method of claim 21 , wherein the AAV capsid encapsulating the first rAAV vector is a serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, or Anc80.
23. The method of claim 21 , wherein the AAV capsid encapsulating the second rAAV vector is a serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, or Anc80.
24. The method of claim 21 , wherein the first rAAV vector is encapsulated by an Anc80 capsid and the second rAAV vector is encapsulated by an Anc80 capsid.
25. The method of claim 24 , wherein the Anc80 capsids comprise the polypeptide sequence of SEQ ID NO: 109.
26. The method of claim 19 , wherein the plurality of rAAV vectors is administered as a single dose.
27. The method of claim 19 , wherein the plurality of rAAV vectors is administered as two or more doses.
28. The method of claim 19 , wherein the subject has a defective otoferlin gene.