Materials and methods for delivering nucleic acids to cochlear and vestibular cells

Patent metadata

Field	Value
Publication number	US-12102692-B2
Application number	US-202117450008-A
Country	US
Kind code	B2
Filing date	Oct 5, 2021
Priority date	Dec 11, 2015
Publication date	Oct 1, 2024
Grant date	Oct 1, 2024

Abstract

Provided herein are materials and methods for efficiently delivering nucleic acids to cochlear and vestibular cells.

First claim

What is claimed is:

1. An adeno-associated virus (AAV) vector comprising an Anc80 capsid protein and a transgene comprising ACTG1, ADCY1, ATOHI, ATP6V1B1, BDNF, BDP1, BSND, DATSPER2, CABP2, CD164, CDC14A, CEACAM16, CHD7, CCDC50, CLDN14, CLIC5, CLPP, COCH, COL2A1, COL4A3, COL4A4, COL4A5, COL9A1, COL9A2, COL11A1, COL11A2, CRYM, DCDC2, DFNA5, DFNB31, DFNB59, DIAPH1, EDN3, EDNRB, ELMOD3, EMOD3, EPS8, EPS8L2, ESPN, ESRRB, EYA1, EYA4, FAM65B, FOXII, GIPC3, GJB2, GJB3, GJB6, GPR98, GRHL2, GPSM2, GRXCR1, GRXCR2, HARS2, HGF, HOMER2, HSD17B4, ILDR1, KARS, KCNE1, KCNJ10, KCNQ1, KCNQ4, KITLG, LARS2, LHFPL5, LOXHD1, LRTOMT, MARVELD2, MCM2, MET, MIR183, MIRN96, MITF, MSRB3, MT-RNR1, MT-TS1, MYH14, MYH9, MYO15A, MYO1A, MY03A, MY06, NARS2, NDP,NF2, NT3, OSBPL2, OTOA, OTOF, OTOG, OTOGL, P2RX2, PAX3, PJVK, PNPT1, POLRID, POLRIC, POU3F4, POU4F3, PRPS1, PTPRQ, RDX, SIPR2, SEMA3E, SERPINB6, SLC17A8, SLC22A4, SLC26A4, SLC26A5, SIX1, SIX5, SMAC/DIABLO, SNAI2, SOX10, STRC, SYNE4, TBC1D24, TCOF1, TECTA, TIMM8A, TJP2, TNC, TMIE, TMEM132E, TMPRSS3, TRPN, TRIOBP, TSPEAR, USHIC, USHIG, USH2D, WFS1, or XIAP.

2. A method of delivering a transgene to at least 80% of inner hair cells (IHCs) and at least 80% of outer ear hair cells (OHCs) in a subject's inner ear to treat a hearing disorder, the method comprising:
administering the AAV vector of claim 1 to the inner ear in a subject.

3. The method of claim 2 , wherein the transgene is further delivered to one or more spiral ganglion neurons, vestibular hair cells, vestibular ganglion neurons, supporting cells, and/or cells in the stria vascularis.

4. The method of claim 2 , wherein the Anc80 capsid protein has the amino acid sequence shown in SEQ ID NO:1.

5. The method of claim 2 , wherein the Anc80 capsid protein has the amino acid sequence shown in SEQ ID NO:2.

6. The method of claim 2 , wherein the transgene is under control of a heterologous promoter sequence.

7. The method of claim 6 , wherein the heterologous promoter sequence comprises a CMV promoter, a CBA promoter, a CASI promoter, a PGK promoter, a EF-1 promoter, an alpha9 nicotinic receptor promoter, a prestin promoter, a KCNQ4 promoter, a Myo7a promoter, a Myo6 promoter, a Gfil promoter, a Vglut3 promoter, or an Atoh1 promoter.

8. The method of claim 2 , wherein the administering step comprises injecting the AAV vector through the round window.

9. The method of claim 2 , wherein the AAV vector is administered during a cochleostomy or during a canalostomy.

10. The method of claim 2 , wherein the AAV vector is administered to the middle ear and/or the round window via one or more drug delivery vehicles.

11. The method of claim 2 , wherein expression of the transgene results in regeneration of inner hair cells (IHCs), or outer hair cells (OHCs), and one or more of spiral ganglion neurons, stria vascularis, vestibular hair cells, and/or vestibular ganglion neurons, thereby restoring hearing or vestibular function.

12. The AAV vector of claim 1 , wherein the Anc80 capsid protein comprises the amino acid sequence of SEQ ID NO:1.

13. The AAV vector of claim 1 , wherein the Anc80 capsid protein comprises the amino acid sequence of SEQ ID NO:2.

14. The AAV vector of claim 1 , wherein the transgene is under control of a heterologous promoter sequence.

15. The AAV vector of claim 14 , wherein the heterologous promoter sequence comprises a CMV promoter, a CBA promoter, a CASI promoter, a PGK promoter, a EF-1 promoter, an alpha9 nicotinic receptor promoter, a prestin promoter, a KCNQ4 promoter, a Myo7a promoter, a Myo6 promoter, a Gfil promoter, a Vglut3 promoter, or an Atoh1 promoter.

16. An adeno-associated virus (AAV) vector comprising an Anc80 capsid protein, and a transgene chosen from KCNQ4, USHIC, GJB2, SLC26A4, STRC, or OTOF.

17. The AAV vector of claim 16 , wherein the Anc80 capsid protein comprises the amino acid sequence of SEQ ID NO: 1.

18. The AAV vector of claim 16 , wherein the Anc80 capsid protein comprises the amino acid sequence of SEQ ID NO:2.

19. The AAV vector of claim 16 , wherein the transgene is under control of a heterologous promoter sequence.

20. The AAV vector of claim 19 , wherein the heterologous promoter sequence comprises a CMV promoter, a CBA promoter, a CASI promoter, a PGK promoter, a EF-1 promoter, an alpha9 nicotinic receptor promoter, a prestin promoter, a KCNQ4 promoter, a Myo7a promoter, a Myo6 promoter, a Gfil promoter, a Vglut3 promoter, or an Atoh1 promoter.

21. The AAV vector of claim 16 , wherein the transgene comprises KCNQ4.

22. The AAV vector of claim 16 , wherein the transgene comprises USHIC.

23. The AAV vector of claim 16 , wherein the transgene comprises GJB2.

24. The AAV vector of claim 16 , wherein the transgene comprises SLC26A4.

25. A method of delivering a transgene to at least 80% of inner hair cells (IHCs) and at least 80% of outer ear hair cells (OHCs) in a subject's inner ear to treat a hearing disorder, the method comprising administering the AAV vector of claim 16 to the inner ear in a subject.

26. The method of claim 25 , wherein the transgene is further delivered to one or more of spiral ganglion neurons, vestibular hair cells, vestibular ganglion neurons, supporting cells, or cells in the stria vascularis.

27. The method of claim 25 , wherein the Anc80 capsid protein has the amino acid sequence shown in SEQ ID NO:1.

28. The method of claim 25 , wherein the Anc80 capsid protein has the amino acid sequence shown in SEQ ID NO:2.

Assignees

• Massachusetts Eye & Ear Infirmary
• Schepens Eye Res Inst
• Childrens Medical Center
• Schepens Eye Res Inst

Inventors

• Stankovic Konstantina
• Vandenberghe Luk H
• Holt Jeffrey
• Geleoc Gwenaelle

Classifications

• A61K9/0046

  Ear · CPC title

• A61K9/0019

  Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner (non-active ingredients are additionally classified in A61K47/00) · CPC title

• C12N2750/14145

  Special targeting system for viral vectors · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title

• C12N2750/14122

  New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes · CPC title