AAV-mediated delivery of antibodies to the inner ear
Provided herein are methods that include introducing into an inner ear of a mammal a therapeutically effective amount of an adeno-associated virus (AAV) vector that includes a nucleotide sequence encoding (a) a polypeptide including an antibody heavy chain variable domain operably linked to a signal peptide and a polypeptide including an antibody light chain variable domain operably linked to a signal peptide; (b) a polypeptide including an antigen-binding antibody fragment operably linked to a signal peptide; or (c) a soluble vascular endothelial growth factor receptor operably linked to a signal peptide.
1. A method of delivering anti-VEGF antibody to an inner ear of a mammal, said method comprising:
delivering via intra-cochlear administration an adeno-associated virus (AAV) vector into the inner ear of the mammal, and
wherein the AAV vector comprises a nucleotide sequence comprising:
(i) a first coding sequence that encodes a first polypeptide comprising an antibody heavy chain variable domain operably linked to a first signal peptide, wherein the first coding sequence comprises the nucleotide sequence of SEQ ID NO: 52;
(ii) a second coding sequence that encodes a second polypeptide comprising an antibody light chain variable domain operably linked to a second signal peptide, wherein the second coding sequence comprises the nucleotide sequence of SEQ ID NO: 53; and
(iii) a sequence encoding a Thosea asigna virus 2A (T2A) peptide that is present between the first coding sequence and the second coding sequence,
wherein the first and second polypeptides together specifically bind to one or more mammalian VEGF proteins,
wherein the mammal has an inner ear disorder and the inner ear disorder is vestibular schwannoma or neurofibromatosis type II, and
wherein the delivery of the AAV vector results in a reduction in VEGF activity in the inner ear of the mammal.
2. The method of claim 1 , wherein the nucleotide sequence comprises one or both of (i) a promoter and (ii) a Kozak sequence.
3. The method of claim 2 , wherein the promoter is an inducible promoter, a constitutive promoter, or a tissue-specific promoter.
4. The method of claim 3 , wherein the constitutive promoter is a CAG promoter, a CBA promoter, or a CMV promoter.
5. The method of claim 1 , wherein the nucleotide sequence comprises a polyadenylation signal sequence.
6. The method of claim 1 , wherein one or both of the first and second signal peptides comprise an IL2 signal peptide.
7. The method of claim 1 , wherein the nucleotide sequence further comprises two AAV inverted terminal repeats (ITRs), wherein the two AAV ITRs flank both of the coding sequences.
8. The method of claim 7 , wherein the two AAV ITRs are AAV2 ITRs or are derived from AAV2 ITRs.
9. The method of claim 1 , wherein the AAV vector is comprised in a composition.
10. The method of claim 1 , wherein intra-cochlear administration comprises delivery into or through a round window membrane.
11. The method of claim 1 , wherein the reduction in VEGF activity is due to binding of VEGF-A, VEGF-B, VEGF-C, and/or VEGF-D by the first polypeptide and the second polypeptide.
12. The method of claim 1 , further comprising indirectly detecting a level of VEGF activity by measuring a hearing level of the mammal.
13. The method of claim 1 , wherein the inner ear disorder is vestibular schwannoma.
14. The method of claim 1 , wherein the mammal is a human.
15. The method of claim 1 , wherein the reduced VEGF activity is reduced human VEGF-A, VEGF-B, VEGF-C, and/or VEGF-D activity.
16. The method of claim 1 , wherein the inner ear disorder is neurofibromatosis type II.