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

Abstract

Provided herein are materials and methods for efficiently delivering nucleic acids to cochlear and vestibular cells, and methods of treating sensory transduction disorders associated with a genetic defect. Some embodiments are directed to a synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector, a cell comprising the synthetic inner ear hair cell targeting AAV vector, and method of treating Usher Syndrome in a subject using the synthetic inner ear hair cell targeting AAV vector.

What is claimed is: 
     
         1 . A synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector, wherein the vector encodes a capsid having at least about 85% sequence identity to Anc80, and comprises a promoter selected from the group consisting of an Espin promoter, a PCDH15 promoter, a PTPRQ promoter and a TMHS (LHFPL5) promoter that directs expression of harmonin-a, harmonin-b, or harmonin-c polypeptide. 
     
     
         2 . The synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector of  claim 1 , wherein the vector transduces inner and outer hair cells with at least about 70% or greater efficiency. 
     
     
         3 . A cell comprising the synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector of  claim 1 . 
     
     
         4 . The cell of  claim 3 , wherein the cell is an outer or inner hair cell. 
     
     
         5 . A method of treating Usher Syndrome in a subject, the method comprising contacting a cell of the subject with the synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector of  claim 1 . 
     
     
         6 . A method for introduction of a wild-type form of a defective gene in a subject with Usher Syndrome, the method comprising contacting a cell of the subject with the synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector of  claim 1 . 
     
     
         7 . A method of treating Usher Syndrome in a subject, the method comprising contacting a cell of the subject with the synthetic inner ear hair cell targeting adeno-associated virus (AAV) vector, wherein the vector comprises a promoter selected from the group consisting of an Espin promoter, a PCDH15 promoter, a PTPRQ promoter and a TMHS (LHFPL5) promoter, wherein the promoter directs expression of polynucleotide encoding a human USH1 polypeptide selected from the group consisting of myosin 7a, harmonin, cadherin 23, protocadherin 15, SANS and calcium and integrin binding protein 2. 
     
     
         8 . The method of  claim 7 , wherein the human polypeptide is TMC1, TMC2, harmonin-a, harmonin-b, or harmonin-c. 
     
     
         9 . The method of  claim 7 , wherein the administering reverses the hearing loss. 
     
     
         10 . The method of  claim 9 , wherein the hearing loss is partial hearing loss or complete deafness. 
     
     
         11 . The method of  claim 9 , wherein recovery of auditory function is associated with preservation of hair bundle morphology and/or restoration of mechanotransduction.