WO2020097372A1 - Use of adeno-associated viral vectors to correct gene defects/ express proteins in hair cells and supporting cells in the inner ear - Google Patents

Use of adeno-associated viral vectors to correct gene defects/ express proteins in hair cells and supporting cells in the inner ear Download PDF

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Publication number
WO2020097372A1
WO2020097372A1 PCT/US2019/060328 US2019060328W WO2020097372A1 WO 2020097372 A1 WO2020097372 A1 WO 2020097372A1 US 2019060328 W US2019060328 W US 2019060328W WO 2020097372 A1 WO2020097372 A1 WO 2020097372A1
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amino acid
acid substitutions
amino acids
cell target
sequence
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French (fr)
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WO2020097372A8 (en
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Emmanuel J. Simons
Robert NG
Danielle LENZ
Michelle VALERO
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Akouos Inc
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Akouos Inc
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Priority to JP2021524060A priority Critical patent/JP2022506613A/ja
Priority to EP19881503.7A priority patent/EP3880826A4/en
Priority to AU2019377115A priority patent/AU2019377115A1/en
Priority to CN201980082439.4A priority patent/CN113853437A/zh
Priority to US17/291,931 priority patent/US20230056182A1/en
Priority to EP22155160.9A priority patent/EP4063510A1/en
Priority to CA3119065A priority patent/CA3119065A1/en
Priority to CN202410531995.8A priority patent/CN118497276A/zh
Application filed by Akouos Inc filed Critical Akouos Inc
Publication of WO2020097372A1 publication Critical patent/WO2020097372A1/en
Publication of WO2020097372A8 publication Critical patent/WO2020097372A8/en
Anticipated expiration legal-status Critical
Priority to JP2024048210A priority patent/JP2024086735A/ja
Priority to US18/806,641 priority patent/US20250041457A1/en
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    • C—CHEMISTRY; METALLURGY
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/0075—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the delivery route, e.g. oral, subcutaneous
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/16—Otologicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/705—Receptors; Cell surface antigens; Cell surface determinants
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/65—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression using markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/03—Phosphoric monoester hydrolases (3.1.3)
    • C12Y301/03001—Alkaline phosphatase (3.1.3.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
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    • C12N2800/00—Nucleic acids vectors
    • C12N2800/40—Systems of functionally co-operating vectors

Definitions

  • the present disclosure relates generally to the use of nucleic acids to treat hearing loss in a primate.
  • Hearing loss can be conductive (arising from the ear canal or middle ear), sensorineural (arising from the inner ear or auditory nerve), or mixed. Most forms of non-syndromic deafness are associated with permanent hearing loss caused by damage to structures in the inner ear (sensorineural deafness), although some forms may involve changes in the middle ear (conductive hearing loss).
  • sensorineural hearing loss is caused by abnormalities in the hair cells of the organ of Corti in the cochlea (poor hair cell ftinction). The hair cells may be abnormal at birth, or may be damaged during the lifetime of an individual (e.g., as a result of noise trauma or infection).
  • each of the at least two different adeno- associated virus (AAV) vectors includes a coding sequence that encodes a different portion of a supporting cell target protein, each of the encoded portions being at least 30 amino acid residues in length, wherein the amino acid sequence of each of the encoded portions may optionally partially overlap with the amino acid sequence of a different one of the encoded portions; no single vector of the at least two different vectors encodes the lull-length supporting cell target protein; at least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of the supporting cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons; and when introduced into a primate cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a lull-
  • AAV adeno- associated virus
  • compositions that include at least two different nucleic acid vectors, where: each of the at least two different adeno-associated virus (AAV) vectors includes a coding sequence that encodes a different portion of a supporting cell target protein, each of the encoded portions being at least 30 amino acid residues in length, where the amino acid sequence of each of the encoded portions may optionally partially overlap with the amino acid sequence of a different one of the encoded portions; no single vector of the at least two different vectors encodes the lull-length supporting cell target protein; at least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of the supporting cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons; and when introduced into a primate cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a lull-length supporting cell target protein.
  • AAV
  • the amino acid sequence of one of the encoded portions overlaps with the amino acid sequence of a different one of the encoded portions. In some embodiments of any of the compositions described herein, the amino acid sequence of each of the encoded portions partially overlaps with the amino acid sequence of a different encoded portion. In some embodiments of any of the compositions described herein, the overlapping amino acid sequence is between 30 amino acid residues to about 390 amino acid residues in length.
  • each of the at least two different vectors includes a different segment of an intron, where the intron includes the nucleotide sequence of an intron that is present in a supporting cell target genomic DNA, and where the two different segments overlap in sequence by at least 100 nucleotides.
  • the two intron includes the nucleotide sequence of an intron that is present in a supporting cell target genomic DNA, and where the two different segments overlap in sequence by at least 100 nucleotides.
  • nucleotide sequence of each of the at least two different vectors is between about 30 nucleotides to about 390 nucleotides in length. In some embodiments of any of the compositions described herein, the nucleotide sequence of each of the at least two different vectors is between 30 nucleotides to 340 nucleotides in length.
  • the number of different vectors in the composition is two.
  • a first of the two different vectors includes a coding sequence that encodes an N-terminal portion of the supporting cell target protein.
  • the N-terminal portion of the supporting cell target protein is between about 30 amino acids to about 390 amino acids in length.
  • the N-terminal portion of the supporting cell target protein is between about 30 amino acids to about 340 amino acids in length.
  • the first vector fiirther includes one or both of a promoter and a Kozak sequence.
  • the first vector includes a promoter that is an inducible promoter, a constitutive promoter, or a tissue-specific promoter.
  • the second of the two different vectors includes a coding sequence that encodes a C-terminal portion of the supporting cell target protein.
  • the C-terminal portion of the supporting cell target protein is between 30 amino acids to about 390 amino acids in length. In some embodiments of any of the compositions described herein, the C-terminal portion of the supporting cell target protein is between 30 amino acids to about 340 amino acids in length.
  • the second vector fiirther includes a poly(dA) sequence.
  • compositions that include two different nucleic acid vectors, where: a first nucleic acid vector of the two different nucleic acid vectors includes a promoter, a first coding sequence that encodes an N-terminal portion of a
  • a second nucleic acid vector of the two different nucleic acid vectors includes a splicing acceptor signal sequence, a second coding sequence that encodes a C-terminal portion of a supporting cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the amino acid sequences of the encoded portions do not overlap, where no single vector of the two different vectors encodes the lull-length supporting cell target protein, and, when the coding sequences are transcribed in a primate cell, to produce RNA transcripts, splicing occurs between the splicing donor signal sequence on one transcript and the splicing acceptor signal sequence on the other transcript, thereby forming a recombine
  • At least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of a supporting cell target genomic DNA, and lacks an intronic sequence between the neighboring exons.
  • compositions that include: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a supporting cell target protein positioned 3’ of the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a first detectable marker gene positioned 3’ of the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second detectable marker gene, a splicing acceptor signal sequence positioned 3’ of the second detectable marker gene, a second coding sequence that encodes a C-terminal portion of a supporting cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence positioned at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the respective amino acid sequences of the encoded portions do not overlap with each other, where no single
  • RNA transcripts splicing occurs between the splicing donor signal on one transcript and the splicing acceptor signal on the other transcript, thereby forming a recombined RNA molecule that encodes a lull-length supporting cell target protein.
  • At least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of a supporting cell target genomic DNA, and lacks an intronic sequence between the neighboring exons.
  • the first or second detectable marker gene is alkaline phosphatase. In some embodiments of any of the compositions described herein, the first and second detectable marker genes are the same.
  • compositions that include: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a supporting cell target protein positioned 3’ to the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a FI phage recombinogenic region positioned 3’ to the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second FI phage recombinogenic region, a splicing acceptor signal sequence positioned 3’ of the second FI phage recombinogenic region, a second coding sequence that encodes a C- terminal portion of a supporting cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence positioned at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the respective
  • At least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of a supporting cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons.
  • compositions that include a single adeno-associated virus (AAV) vector, where the single AAV vector that includes a nucleic acid sequence that encodes a supporting cell target protein; and when introduced into a primate cell, a nucleic acid encoding a lull-length supporting cell target protein is generated at the locus of the supporting cell target gene, and the primate expresses the supporting cell target protein.
  • AAV adeno-associated virus
  • the supporting cell target gene is gap junction protein beta 2 (GJB2). In some embodiments of any of the compositions described herein, the supporting cell target gene is solute carrier family 26 member 4 (SLC26A4).
  • the composition farther includes a pharmaceutically acceptable excipient.
  • the second nucleic acid vector lurther includes a destabilizing sequence.
  • the second nucleic acid vector lurther includes a FKBP12 destabilizing sequence.
  • kits including any of the compositions described herein.
  • the kit lurther includes a pre- loaded syringe containing the composition.
  • Also provided herein are methods of correcting a supporting cell target gene defect in a supporting cell of an inner ear in a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering repairs the supporting cell target gene defect in the supporting cell of the inner ear of the primate.
  • Also provided herein are methods of increasing the expression level of a supporting cell target protein in a supporting cell of an inner ear of a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering results in an increase in the expression level of the supporting cell target protein in the supporting cell of the inner ear of the primate.
  • the primate has previously been determined to have a defective supporting cell target gene.
  • Also provided herein are methods of treating non-syndromic sensorineural hearing loss in a primate identified as having a defective supporting cell target gene that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein.
  • the method fiirther includes, prior to the administering step, determining that the primate has a defective supporting cell target gene.
  • the method fiirther includes prior to the administering step, determining that the primate has a defective supporting cell target gene.
  • Also provided herein are methods of restoring synapses and/or preserving spiral ganglion nerves in a primate identified or diagnosed as having an inner ear disorder that include: administering to the inner ear of the primate a therapeutically effective amount of any composition described herein.
  • the method fiirther includes prior to the administering step, determining that the primate has a defective supporting cell target gene.
  • Also provided herein are methods including administering to an inner ear of a primate a therapeutically effective amount of any composition described herein.
  • the primate has been previously identified as having a defective supporting cell target gene.
  • compositions including at least two different nucleic acid vectors, wherein: each of the at least two different adeno-associated virus (AAV) vectors includes a coding sequence that encodes a different portion of a hair cell target
  • each of the encoded portions being at least 30 amino acid residues in length, wherein the amino acid sequence of each of the encoded portions may optionally partially overlap with the amino acid sequence of a different one of the encoded portions; no single vector of the at least two different vectors encodes the lull-length hair cell target protein; at least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of the hair cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons; and when introduced into a primate cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a lull-length hair cell target protein.
  • compositions that include at least two different nucleic acid vectors, where: each of the at least two different adeno-associated virus (AAV) vectors includes a coding sequence that encodes a different portion of a hair cell target protein, each of the encoded portions being at least 30 amino acid residues in length, where the amino acid sequence of each of the encoded portions may optionally partially overlap with the amino acid sequence of a different one of the encoded portions; no single vector of the at least two different vectors encodes the lull-length protein; at least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of the hair cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons; and when introduced into a primate cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a lull-length hair cell target protein.
  • AAV adeno
  • the amino acid sequence of one of the encoded portions overlaps with the amino acid sequence of a different one of the encoded portions. In some embodiments of any of the compositions described herein, the amino acid sequence of each of the encoded portions partially overlaps with the amino acid sequence of a different encoded portion. In some embodiments of any of the compositions described herein, the overlapping amino acid sequence is between 30 amino acid residues to about 390 amino acid residues in length.
  • each of the at least two different vectors includes a different segment of an intron, where the intron includes the nucleotide sequence of an intron that is present in a hair cell target genomic DNA, and where the two different segments overlap in sequence by at least 100 nucleotides. In some embodiments of any of the compositions described herein, the two different segments overlap in sequence by about 30 nucleotides to about 390 nucleotides. In some embodiments of any of the compositions described herein, the nucleotide sequence of each of the at least two different vectors is between about 30 nucleotides to about 390 nucleotides in length. In some embodiments of any of the compositions described herein, the nucleotide sequence of each of the at least two different vectors is between 30 nucleotides to 340 nucleotides in length.
  • the number of different vectors in the composition is two.
  • a first of the two different vectors includes a coding sequence that encodes an N-terminal portion of the hair cell target protein.
  • the N-terminal portion of the hair cell target protein is between about 30 amino acids to about 390 amino acids in length.
  • the N- terminal portion of the hair cell target protein is between about 30 amino acids to about 340 amino acids in length.
  • the first vector fiirther includes one or both of a promoter and a Kozak sequence.
  • the first vector includes a promoter that is an inducible promoter, a constitutive promoter, or a tissue- specific promoter.
  • the second of the two different vectors includes a coding sequence that encodes a C-terminal portion of the hair cell target protein.
  • the C-terminal portion of the hair cell target protein is between 30 amino acids to about 390 amino acids in length. In some embodiments of any of the compositions described herein, the C-terminal portion of the hair cell target protein is between 30 amino acids to about 340 amino acids in length.
  • the second vector fiirther includes a poly(dA) sequence.
  • compositions that include two different nucleic acid vectors, where: a first nucleic acid vector of the two different nucleic acid vectors includes a promoter, a first coding sequence that encodes an N-terminal portion of a hair cell target protein positioned 3’ of the promoter, and a splicing donor signal sequence positioned at the 3’ end of the first coding sequence; and a second nucleic acid vector of the two different nucleic acid vectors includes a splicing acceptor signal sequence, a second coding sequence that encodes a C-terminal portion of a hair cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the amino acid sequences of the encoded portions do not overlap, where no single vector of the two different vectors encodes the lull-length hair cell target protein, and, when the coding
  • At least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of a hair cell target genomic DNA, and lacks an intronic sequence between the neighboring exons.
  • compositions that include: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a hair cell target protein positioned 3’ of the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a first detectable marker gene positioned 3’ of the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second detectable marker gene, a splicing acceptor signal sequence positioned 3’ of the second detectable marker gene, a second coding sequence that encodes a C-terminal portion of a hair cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation
  • each of the encoded portions is at least 30 amino acid residues in length, where the respective amino acid sequences of the encoded portions do not overlap with each other, where no single vector of the two different vectors encodes the lull-length hair cell target protein, and, when the coding sequences are transcribed in a primate cell to produce RNA transcripts, splicing occurs between the splicing donor signal on one transcript and the splicing acceptor signal on the other transcript, thereby forming a recombined RNA molecule that encodes a lull-length hair cell target protein.
  • At least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of a hair cell target genomic DNA, and lacks an intronic sequence between the neighboring exons.
  • the first or second detectable marker gene is alkaline phosphatase. In some embodiments of any of the compositions described herein, the first and second detectable marker genes are the same.
  • compositions that include: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a hair cell target protein positioned 3’ to the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a FI phage recombinogenic region positioned 3’ to the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second FI phage recombinogenic region, a splicing acceptor signal sequence positioned 3’ of the second FI phage recombinogenic region, a second coding sequence that encodes a C-terminal portion of a hair cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence positioned at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the
  • At least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of a hair cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons.
  • compositions that include a single adeno-associated virus (AAV) vector, where the single AAV vector that includes a nucleic acid sequence that encodes a hair cell target protein; and when introduced into a primate cell, a nucleic acid encoding a lull-length hair cell target protein is generated at the locus of the hair cell target gene, and the primate expresses the hair cell target protein.
  • AAV adeno-associated virus
  • the composition lurther includes a pharmaceutically acceptable excipient.
  • kits including any of the compositions described herein.
  • the kit lurther includes a pre- loaded syringe containing the composition.
  • Also provided herein are methods of correcting a hair cell target gene defect in a hair cell of an inner ear in a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering repairs the hair cell target gene defect in the hair cell of the inner ear of the primate.
  • Also provided herein are methods of increasing the expression level of a hair cell target protein in a hair cell of an inner ear of a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering results in an increase in the expression level of the hair cell target protein in the hair cell of the inner ear of the primate.
  • the primate has previously been determined to have a defective hair cell target gene.
  • Also provided herein are methods of treating non-syndromic sensorineural hearing loss in a primate identified as having a defective hair cell target gene that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein.
  • the method lurther includes, prior to the administering step, determining that the primate has a defective hair cell target gene.
  • the method lurther includes prior to the administering step, determining that the primate has a defective hair cell target gene.
  • Also provided herein are methods of restoring synapses and/or preserving spiral ganglion nerves in a primate identified or diagnosed as having an inner ear disorder that include: administering to the inner ear of the primate a therapeutically effective amount of any composition described herein.
  • the method lurther includes prior to the administering step, determining that the primate has a defective hair cell target gene.
  • Also provided herein are methods including administering to an inner ear of a primate a therapeutically effective amount of any composition described herein.
  • the primate has been previously identified as having a defective hair cell target gene.
  • “a” and“an” refers to one or to more than one (i.e., at least one) of the grammatical object of the article.
  • “an element” encompasses one element and more than one element.
  • supporting cell target protein means a protein that is expressed in a supporting cell in a primate and ftinctionally contributes, at least in part, to the hearing in the primate.
  • supporting cell target proteins include GJB2 and SLC26A4.
  • mutation in a supporting cell target gene refers to a modification in a wildtype supporting cell target gene that results in the production of a supporting cell
  • a mutation can result in the production of a supporting cell target protein having a deletion in one or more amino acids (e.g., 2, 3, 4,
  • the mutation can result in a frameshift in the supporting cell target gene.
  • hair cell target protein means a protein that is expressed in a hair cell in a primate and ftinctionally contributes, at least in part, to the hearing in the primate.
  • hair cell target proteins include tectorin alpha (TECTA), unconventional myson-15 (MY015A), myosin IILA (MY03A), myosin VI (MY06), myosin VILA (MY07A), cadherin related 23 (CDH23), collagen type XI alpha 2 chain (COL11A2), lipoxygenase homology domains 1 (LOXHD1), protocadherin related 15 (PCDH15), whirlin (WHRN), an otoferlin (OTOF) gene, a trio rho guanine nucleotide exchange factor (TRIO) and f-actin binding protein (TRIOBP) gene, a stereocilin (STRC) gene, a pejvakin (PJVK) gene, a norrin
  • TECTA
  • mutation in a hair cell target gene refers to a modification in a wildtype hair cell target gene that results in the production of a hair cell target protein having one or more of: a deletion in one or more amino acids, one or more amino acid substitutions, and one or more amino acid insertions as compared to the wildtype hair cell target protein, and/or results in a decrease in the expressed level of the encoded hair cell target protein in a primate cell as compared to the expressed level of the encoded hair cell target protein in a primate cell not having a mutation.
  • a mutation can result in the production of a hair cell target protein having a deletion in one or more amino acids (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 16, 17, 18, 19, or 20 amino acids). In some embodiments, the mutation can result in a frameshift in the hair cell
  • a frameshift is known in the art to encompass any mutation in a coding sequence that results in a shift in the reading frame of the coding sequence.
  • a frameshift can result in a nonfunctional protein.
  • a point mutation can be a nonsense mutation (i.e., result in a premature stop codon in an exon of the gene).
  • a nonsense mutation can result in the production of a truncated protein (as compared to a corresponding wildtype protein) that may or may not be functional.
  • the mutation can result in the loss (or a decrease in the level) of expression of hair cell target mRNA or hair cell target protein or both the mRNA and protein.
  • the mutation can result in the production of an altered hair cell target protein having a loss or decrease in one or more biological activities (functions) as compared to a wildtype hair cell target protein. In some embodiments, the mutation can result in the loss (or a decrease in the level) of expression of supporting cell target mRNA or supporting cell target protein or both the mRNA and protein. In some embodiments, the mutation can result in the production of an altered supporting cell target protein having a loss or decrease in one or more biological activities (functions) as compared to a wildtype supporting cell target protein.
  • the mutation is an insertion of one or more nucleotides into a hair cell target gene.
  • the mutation is in a regulatory sequence of the hair cell target gene, i.e., a portion of the gene that is not coding sequence.
  • a mutation in a regulatory sequence may be in a promoter or enhancer region and prevent or reduce the proper transcription of the hair cell target gene.
  • the mutation is an insertion of one or more nucleotides into a supporting cell target gene.
  • the mutation is in a regulatory sequence of the supporting cell target gene, i.e., a portion of the gene that is not coding sequence.
  • a mutation in a regulatory sequence may be in a promoter or enhancer region and prevent or reduce the proper transcription of the supporting cell target gene.
  • the term“conservative mutation” refers to a mutation that does not change the amino acid encoded at the site of the mutation (due to codon degeneracy).
  • Modifications can be introduced into a nucleotide sequence by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated
  • Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain.
  • Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid and glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, and methionine), beta-branched side chains (e.g., threonine, valine, and isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine
  • nucleotide sequence encoding an amino acid sequence includes all nucleotide sequences that are degenerate versions of each other and thus encode the same amino acid sequence.
  • endogenous refers to any material originating from within an organism, cell, or tissue.
  • exogenous refers to any material introduced from or originating from outside an organism, cell, or tissue that is not produced or does not originate from the same organism, cell, or tissue in which it is being introduced.
  • isolated means altered or removed from the natural state.
  • a nucleic acid or a peptide naturally present in a living animal is not“isolated,” but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its natural state is“isolated.”
  • An isolated nucleic acid or protein can exist in substantially purified form, or can exist in a non-native environment such as, for example, a host cell.
  • transfected refers to a process by which exogenous nucleic acid is transferred or introduced into a cell.
  • A“transfected,” “transformed,” or “transduced” primate cell is one that has been transfected, transformed or transduced with exogenous nucleic acid.
  • the term“expression” refers to the transcription and/or translation of a particular nucleotide sequence encoding a protein.
  • the term“transient expression” refers to the expression of a non-integrated coding sequence for a short period of time (e.g., hours or days).
  • the coding sequence that is transiently expressed in a cell e.g., a primate cell is lost upon multiple rounds of cell division.
  • the term“primate” is intended to include any primate (e.g., a human, a non human primate (e.g., simian (e.g., a monkey (e.g., a marmoset, a baboon, or a macaque), or an ape (e.g., a gorilla, a gibbon, an orangutan, or a chimpanzee).
  • the primate has or is at risk of having hearing loss.
  • the primate has been previously identified as having a mutation in a hair cell target gene (e.g., an OTOF gene, a TRIOBP gene, a STRC gene, a PJVK gene, a NDP gene, a CLRN1 gene, a TMC1 gene, a VEGF gene, or a FISPA1A gene).
  • a hair cell target gene e.g., an OTOF gene, a TRIOBP gene, a STRC gene, a PJVK gene, a NDP gene, a CLRN1 gene, a TMC1 gene, a VEGF gene, or a FISPA1A gene.
  • the primate has been previously identified as having a mutation in a supporting cell target gene (e.g., a GJB2 gene or a SLC26A4 gene).
  • the primate has been previously identified as having a mutation in a GJB2 gene.
  • the primate has been previously identified as having a mutation in a SLC26A4 gene.
  • the primate has been identified as having a mutation in a hair cell target gene (e.g., an OTOF gene, a TRIOBP gene, a STRC gene, a PJVK gene, a NDP gene, a CLRN1 gene, a TMC1 gene, a VEGF gene, or a HSPA1A gene) and has been diagnosed with hearing loss.
  • the primate has been identified as having a mutation in a supporting cell target gene (e.g., a GBJ2 gene or a SLC26A4 gene) and has been diagnosed with hearing loss.
  • the primate has been identified as having hearing loss.
  • a treatment is“therapeutically effective” when it results in a reduction in one or more of the number, severity, and frequency of one or more symptoms of a disease state (hearing loss, e.g., non-syndromic sensorineural hearing loss or syndromic sensorineural hearing loss) in a primate.
  • a disease state e.g., non-syndromic sensorineural hearing loss or syndromic sensorineural hearing loss
  • a therapeutically effective amount of a composition can result in an increase in the expression level of an active target protein (e.g., a supporting cell target protein (e.g., a wildtype, lull-length supporting cell target protein or a variant of a supporting cell target protein that has the desired activity) (e.g., as compared to the expression level prior to treatment with the composition) or a hair cell target protein (e.g., a wildtype, lull-length hair cell target protein or a variant of a hair cell
  • an active target protein e.g., a supporting cell target protein (e.g., a wildtype, lull-length supporting cell target protein or a variant of a supporting cell target protein that has the desired activity)
  • a hair cell target protein e.g., a wildtype, lull-length hair cell target protein or a variant of a hair cell
  • a therapeutically effective amount of a composition can result in an increase in the expression level of an active target protein (e.g., an active supporting cell target protein (e.g., a wildtype, lull-length supporting cell target protein or active variant) ort an active hair cell target protein (e.g., a wildtype, lull-length hair cell target protein or active variant) in a target cell (e.g., a supporting cell or a hair cell).
  • an active target protein e.g., an active supporting cell target protein (e.g., a wildtype, lull-length supporting cell target protein or active variant)
  • an active hair cell target protein e.g., a wildtype, lull-length hair cell target protein or active variant
  • a therapeutically effective amount of a composition can result in an increase in the expression level of an active target protein (e.g., an active supporting cell target protein (e.g., a wildtype, lull-length supporting cell target protein or active variant) or an active hair cell target protein (e.g., a wildtype, lull-length hair cell target protein or active variant), and/or an increase in one or more activities of a target protein (e.g., a supporting cell target protein or a hair cell target protein) in a target cell (e.g., as compared to a reference level, such as the level(s) in a primate prior to treatment, the level(s) in a primate having a mutation in a target gene (e.g., a supporting cell target gene or a hair cell target gene), or the level(s) in a primate or a population of primates having hearing loss, e.g., non-syndromic sensorineural hearing loss or syndromic sensorine
  • nucleic acid or“polynucleotide” refers to deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), or a combination thereof, in either single- or double- stranded form. Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleotides. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses complementary sequences as well as the sequence explicitly indicated. In some embodiments of any of the nucleic acids described herein, the nucleic acid is DNA. In some embodiments of any of the nucleic acids described herein, the nucleic acid is RNA.
  • active hair cell target protein means a protein encoded by DNA that, if substituted for both wildtype alleles encoding lull-length hair cell target protein in hair cells of what is otherwise a wildtype primate, and if expressed in the hair cells of that primate, results in that primate’s having a level of hearing approximating the normal level of hearing of a similar primate that is entirely wildtype.
  • active hair cell target protein means a protein encoded by DNA that, if substituted for both wildtype alleles encoding lull-length hair cell target protein in hair cells of what is otherwise a wildtype primate, and if expressed in the hair cells of that primate, results in that primate’s having a level of hearing approximating the normal level of hearing of a similar primate that is entirely wildtype.
  • hair cell target proteins are lull-length hair cell target proteins (e.g., any of the lull-length hair cell target proteins described herein).
  • active supporting cell target protein means a protein encoded by DNA that, if substituted for both wildtype alleles encoding lull-length supporting cell target protein in supporting cells of what is otherwise a wildtype primate, and if expressed in the supporting cells of that primate, results in that primate’s having a level of hearing approximating the normal level of hearing of a similar primate that is entirely wildtype.
  • active supporting cell target proteins are lull-length supporting cell target proteins (e.g., any of the lull-length supporting cell target proteins described herein).
  • an active target protein e.g., an active supporting cell target protein or an active hair cell target protein
  • can include a sequence of a wildtype, lull-length target protein e.g., a supporting cell target protein (e.g., a wildtype, human, lull-length supporting cell target protein) or a hair cell target protein (e.g., a wildtype, human, lull- length hair cell target protein) including 1 amino acid substitution to about 160 amino acid substitutions, 1 amino acid substitution to about 155 amino acid substitutions, 1 amino acid substitution to about 150 amino acid substitutions, 1 amino acid substitution to about 145 amino acid substitutions, 1 amino acid substitution to about 140 amino acid substitutions, 1 amino acid substitution to about 135 amino acid substitutions, 1 amino acid substitution to about 130 amino acid substitutions, 1 amino acid substitution to about 125 amino acid substitutions, 1 amino acid substitution to about 120 amino acid substitutions, 1 amino acid substitution to about 115 amino acid substitutions, 1 amino acid substitution to about 110 amino acid substitutions, 1 amino acid substitution
  • amino acid substitution to about 50 amino acid substitutions 1 amino acid substitution to about 45 amino acid substitutions, 1 amino acid substitution to about 40 amino acid substitutions, 1 amino acid substitution to about 35 amino acid substitutions, 1 amino acid substitution to about 30 amino acid substitutions, 1 amino acid substitution to about 25 amino acid substitutions, 1 amino acid substitution to about 20 amino acid substitutions, 1 amino acid substitution to about 15 amino acid substitutions, 1 amino acid substitution to about 10 amino acid substitutions, 1 amino acid substitution to about 9 amino acid substitutions, 1 amino acid substitution to about 8 amino acid substitutions,
  • 1 amino acid substitution to about 7 amino acid substitutions 1 amino acid substitution to about 6 amino acid substitutions, 1 amino acid substitution to about 5 amino acid substitutions, 1 amino acid substitution to about 4 amino acid substitutions, 1 amino acid substitution to about 3 amino acid substitutions, between about 2 amino acid substitutions to about 160 amino acid substitutions, about 2 amino acid substitutions to about 155 amino acid substitutions, about 2 amino acid substitutions to about 150 amino acid substitutions, about 2 amino acid substitutions to about 145 amino acid substitutions, about 2 amino acid substitutions to about 140 amino acid substitutions, about 2 amino acid substitutions to about 135 amino acid substitutions, about 2 amino acid substitutions to about 130 amino acid substitutions, about 2 amino acid substitutions to about 125 amino acid substitutions, about 2 amino acid substitutions to about 120 amino acid substitutions, about 2 amino acid substitutions to about 115 amino acid substitutions, about 2 amino acid substitutions to about 110 amino acid substitutions, about 2 amino acid substitutions to about 105 amino acid substitutions, about 2 amino acid substitutions to about 100 amino acid substitutions, about 2 amino acid substitutions
  • amino acid substitutions about 2 amino acid substitutions to about 40 amino acid substitutions, about 2 amino acid substitutions to about 35 amino acid substitutions, about 2 amino acid substitutions to about 30 amino acid substitutions, about 2 amino acid substitutions to about 25 amino acid substitutions, about 2 amino acid substitutions to about 20 amino acid substitutions, about 2 amino acid substitutions to about 15 amino acid substitutions, about 2 amino acid substitutions to about 10 amino acid substitutions, about 2 amino acid substitutions to about 9 amino acid substitutions, about 2 amino acid substitutions to about 8 amino acid substitutions, about 2 amino acid substitutions to about 7 amino acid substitutions, about 2 amino acid substitutions to about 6 amino acid substitutions, about 2 amino acid substitutions to about 5 amino acid substitutions, about 2 amino acid substitutions to about 4 amino acid substitutions, between about 3 amino acid substitutions to about 160 amino acid substitutions, about 3 amino acid substitutions to about 155 amino acid substitutions, about 3 amino acid substitutions to about 150 amino acid substitutions, about 3 amino acid substitutions to about 145 amino acid substitutions, about 3 amino acid substitutions to about 140
  • amino acid substitutions to about 35 amino acid substitutions about 3 amino acid substitutions to about 30 amino acid substitutions, about 3 amino acid substitutions to about 25 amino acid substitutions, about 3 amino acid substitutions to about 20 amino acid substitutions, about 3 amino acid substitutions to about 15 amino acid substitutions, about 3 amino acid substitutions to about 10 amino acid substitutions, about 3 amino acid substitutions to about 9 amino acid substitutions, about 3 amino acid substitutions to about 8 amino acid substitutions, about 3 amino acid substitutions to about 7 amino acid substitutions, about 3 amino acid substitutions to about 6 amino acid substitutions, about 3 amino acid substitutions to about 5 amino acid substitutions, between about 4 amino acid substitutions to about 160 amino acid substitutions, about 4 amino acid substitutions to about 155 amino acid substitutions, about 4 amino acid substitutions to about 150 amino acid substitutions, about 4 amino acid substitutions to about 145 amino acid substitutions, about 4 amino acid substitutions to about 140 amino acid substitutions, about 4 amino acid substitutions to about 135 amino acid substitutions, about 4 amino acid substitutions to about 130 amino acid substitutions
  • substitutions to about 25 amino acid substitutions about 4 amino acid substitutions to about 20 amino acid substitutions, about 4 amino acid substitutions to about 15 amino acid substitutions, about 4 amino acid substitutions to about 10 amino acid substitutions, about 4 amino acid substitutions to about 9 amino acid substitutions, about 4 amino acid substitutions to about 8 amino acid substitutions, about 4 amino acid substitutions to about 7 amino acid substitutions, about 4 amino acid substitutions to about 6 amino acid substitutions, between about 5 amino acid substitutions to about 160 amino acid substitutions, about 5 amino acid substitutions to about 155 amino acid substitutions, about 5 amino acid substitutions to about 150 amino acid substitutions, about 5 amino acid substitutions to about 145 amino acid substitutions, about 5 amino acid substitutions to about 140 amino acid substitutions, about 5 amino acid substitutions to about 135 amino acid substitutions, about 5 amino acid substitutions to about 130 amino acid substitutions, about 5 amino acid substitutions to about 125 amino acid substitutions, about 5 amino acid substitutions to about 120 amino acid substitutions, about 5 amino acid substitutions to about 115 amino acid substitutions, about
  • substitutions to about 10 amino acid substitutions about 5 amino acid substitutions to about 9 amino acid substitutions, about 5 amino acid substitutions to about 8 amino acid substitutions, about 5 amino acid substitutions to about 7 amino acid substitutions, between about 6 amino acid substitutions to about 160 amino acid substitutions, about 6 amino acid substitutions to about 155 amino acid substitutions, about 6 amino acid substitutions to about 150 amino acid substitutions, about 6 amino acid substitutions to about 145 amino acid substitutions, about 6 amino acid substitutions to about 140 amino acid substitutions, about 6 amino acid substitutions to about 135 amino acid substitutions, about 6 amino acid substitutions to about 130 amino acid substitutions, about 6 amino acid substitutions to about 125 amino acid substitutions, about 6 amino acid substitutions to about 120 amino acid substitutions, about 6 amino acid substitutions to about 115 amino acid substitutions, about 6 amino acid substitutions to about 110 amino acid substitutions, about 6 amino acid substitutions to about 105 amino acid substitutions, about 6 amino acid substitutions to about 100 amino acid substitutions, about 6 amino acid substitutions to about 95 amino acid substitutions,
  • substitutions to about 155 amino acid substitutions about 7 amino acid substitutions to about 150 amino acid substitutions, about 7 amino acid substitutions to about 145 amino acid substitutions, about 7 amino acid substitutions to about 140 amino acid substitutions, about 7 amino acid substitutions to about 135 amino acid substitutions, about 7 amino acid substitutions to about 130 amino acid substitutions, about 7 amino acid substitutions to about 125 amino acid substitutions, about 7 amino acid substitutions to about 120 amino acid substitutions, about 7 amino acid substitutions to about 115 amino acid substitutions, about 7 amino acid substitutions to about 110 amino acid substitutions, about 7 amino acid substitutions to about 105 amino acid substitutions, about 7 amino acid substitutions to about 100 amino acid substitutions, about 7 amino acid substitutions to about 95 amino acid substitutions, about 7 amino acid substitutions to about 90 amino acid substitutions, about 7 amino acid substitutions to about 85 amino acid substitutions, about 7 amino acid substitutions to about 80 amino acid substitutions, about 7 amino acid substitutions to about 75 amino acid substitutions, about 7 amino acid substitutions to about 70 amino acid substitutions, about
  • amino acid substitutions to about 125 amino acid substitutions about 8 amino acid substitutions to about 120 amino acid substitutions, about 8 amino acid substitutions to about 115 amino acid substitutions, about 8 amino acid substitutions to about 110 amino acid substitutions, about 8 amino acid substitutions to about 105 amino acid substitutions, about 8 amino acid substitutions to about 100 amino acid substitutions, about 8 amino acid substitutions to about 95 amino acid substitutions, about 8 amino acid substitutions to about 90 amino acid substitutions, about 8 amino acid substitutions to about 85 amino acid substitutions, about 8 amino acid substitutions to about 80 amino acid substitutions, about 8 amino acid substitutions to about 75 amino acid substitutions, about 8 amino acid substitutions to about 70 amino acid substitutions, about 8 amino acid substitutions to about 65 amino acid substitutions, about 8 amino acid substitutions to about 60 amino acid substitutions, about 8 amino acid substitutions to about 55 amino acid substitutions, about 8 amino acid substitutions to about 50 amino acid substitutions, about 8 amino acid substitutions to about 45 amino acid substitutions, about 8 amino acid substitutions to about 40 amino acid substitutions, about 8
  • amino acid substitutions to about 90 amino acid substitutions about 10 amino acid substitutions to about 85 amino acid substitutions, about 10 amino acid substitutions to about 80 amino acid substitutions, about 10 amino acid substitutions to about 75 amino acid substitutions, about 10 amino acid substitutions to about 70 amino acid substitutions, about 10 amino acid substitutions to about 65 amino acid substitutions, about 10 amino acid substitutions to about 60 amino acid substitutions, about 10 amino acid substitutions to about 55 amino acid substitutions, about 10 amino acid substitutions to about 50 amino acid substitutions, about 10 amino acid substitutions to about 45 amino acid substitutions, about 10 amino acid substitutions to about 40 amino acid substitutions, about 10 amino acid substitutions to about 35 amino acid substitutions, about 10 amino acid substitutions to about 30 amino acid substitutions, about 10 amino acid substitutions to about 25 amino acid substitutions, about 10 amino acid substitutions to about 20 amino acid substitutions, about 10 amino acid substitutions to about 15 amino acid substitutions, between about 15 amino acid substitutions to about 160 amino acid substitutions, about 15 amino acid substitutions to about 155 amino acid substitutions, about
  • amino acid substitutions about 15 amino acid substitutions to about 50 amino acid substitutions, about 15 amino acid substitutions to about 45 amino acid substitutions, about 15 amino acid substitutions to about 40 amino acid substitutions, about 15 amino acid substitutions to about 35 amino acid substitutions, about 15 amino acid substitutions to about 30 amino acid substitutions, about 15 amino acid substitutions to about 25 amino acid substitutions, about 15 amino acid substitutions to about 20 amino acid substitutions, between about 20 amino acid substitutions to about 160 amino acid substitutions, about 20 amino acid substitutions to about 155 amino acid substitutions, about 20 amino acid substitutions to about 150 amino acid substitutions, about 20 amino acid substitutions to about 145 amino acid substitutions, about 20 amino acid substitutions to about 140 amino acid substitutions, about 20 amino acid substitutions to about 135 amino acid substitutions, about 20 amino acid substitutions to about 130 amino acid substitutions, about 20 amino acid substitutions to about 125 amino acid substitutions, about 20 amino acid substitutions to about 120 amino acid substitutions, about 20 amino acid substitutions to about 115 amino acid substitutions, about 20 amino acid substitutions
  • substitutions to about 150 amino acid substitutions about 25 amino acid substitutions to about 145 amino acid substitutions, about 25 amino acid substitutions to about 140 amino acid substitutions, about 25 amino acid substitutions to about 135 amino acid substitutions, about 25 amino acid substitutions to about 130 amino acid substitutions, about 25 amino acid substitutions to about 125 amino acid substitutions, about 25 amino acid substitutions to about 120 amino acid substitutions, about 25 amino acid substitutions to about 115 amino acid substitutions, about 25 amino acid substitutions to about 110 amino acid substitutions, about 25 amino acid substitutions to about 105 amino acid substitutions, about 25 amino acid substitutions to about 100 amino acid substitutions, about 25 amino acid substitutions to about 95 amino acid substitutions, about 25 amino acid substitutions to about 90 amino acid substitutions, about 25 amino acid substitutions to about 85 amino acid substitutions, about 25 amino acid substitutions to about 80 amino acid substitutions, about 25 amino acid substitutions to about 75 amino acid substitutions, about 25 amino acid substitutions to about 70 amino acid substitutions, about 25 amino acid substitutions to about 65 amino acid substitutions, about 25
  • substitutions to about 100 amino acid substitutions about 30 amino acid substitutions to about 95 amino acid substitutions, about 30 amino acid substitutions to about 90 amino acid substitutions, about 30 amino acid substitutions to about 85 amino acid substitutions, about 30 amino acid substitutions to about 80 amino acid substitutions, about 30 amino acid substitutions to about 75 amino acid substitutions, about 30 amino acid substitutions to about 70 amino acid substitutions, about 30 amino acid substitutions to about 65 amino acid substitutions, about 30 amino acid substitutions to about 60 amino acid substitutions, about 30 amino acid substitutions to about 55 amino acid substitutions, about 30 amino acid substitutions to about 50 amino acid substitutions, about 30 amino acid substitutions to about 45 amino acid substitutions, about 30 amino acid substitutions to about 40 amino acid substitutions, about 30 amino acid substitutions to about 35 amino acid substitutions, between about 35 amino acid substitutions to about 160 amino acid substitutions, about 35 amino acid substitutions to about 155 amino acid substitutions, about 35 amino acid substitutions to about 150 amino acid substitutions, about 35 amino acid substitutions to about 145 amino acid substitutions, about 35 amino acid
  • amino acid substitutions to about 40 amino acid substitutions between about 40 amino acid substitutions to about 160 amino acid substitutions, about 40 amino acid substitutions to about 155 amino acid substitutions, about 40 amino acid substitutions to about 150 amino acid substitutions, about 40 amino acid substitutions to about 145 amino acid substitutions, about 40 amino acid substitutions to about 140 amino acid substitutions, about 40 amino acid substitutions to about 135 amino acid substitutions, about 40 amino acid substitutions to about 130 amino acid substitutions, about 40 amino acid substitutions to about 125 amino acid substitutions, about 40 amino acid substitutions to about 120 amino acid substitutions, about 40 amino acid substitutions to about 115 amino acid substitutions, about 40 amino acid substitutions to about 110 amino acid substitutions, about 40 amino acid substitutions to about 105 amino acid substitutions, about 40 amino acid substitutions to about 100 amino acid substitutions, about 40 amino acid substitutions to about 95 amino acid substitutions, about 40 amino acid substitutions to about 90 amino acid substitutions, about 40 amino acid substitutions to about 85 amino acid substitutions, about 40 amino acid substitutions to about 80 amino
  • substitutions about 45 amino acid substitutions to about 100 amino acid substitutions, about 45 amino acid substitutions to about 95 amino acid substitutions, about 45 amino acid substitutions to about 90 amino acid substitutions, about 45 amino acid substitutions to about 85 amino acid substitutions, about 45 amino acid substitutions to about 80 amino acid substitutions, about 45 amino acid substitutions to about 75 amino acid substitutions, about 45 amino acid substitutions to about 70 amino acid substitutions, about 45 amino acid substitutions to about 65 amino acid substitutions, about 45 amino acid substitutions to about 60 amino acid substitutions, about 45 amino acid substitutions to about 55 amino acid substitutions, about 45 amino acid substitutions to about 50 amino acid substitutions, between about 50 amino acid substitutions to about 160 amino acid substitutions, about 50 amino acid substitutions to about 155 amino acid substitutions, about 50 amino acid substitutions to about 150 amino acid substitutions, about 50 amino acid substitutions to about 145 amino acid substitutions, about 50 amino acid substitutions to about 140 amino acid substitutions, about 50 amino acid substitutions to about 135 amino acid substitutions, about 50 amino acid substitutions to about 130 amino
  • amino acid substitutions to about 140 amino acid substitutions about 60 amino acid substitutions to about 135 amino acid substitutions, about 60 amino acid substitutions to about 130 amino acid substitutions, about 60 amino acid substitutions to about 125 amino acid substitutions, about 60 amino acid substitutions to about 120 amino acid substitutions, about 60 amino acid substitutions to about 115 amino acid substitutions, about 60 amino acid substitutions to about 110 amino acid substitutions, about 60 amino acid substitutions to about 105 amino acid substitutions, about 60 amino acid substitutions to about 100 amino acid substitutions, about 60 amino acid substitutions to about 95 amino acid substitutions, about 60 amino acid substitutions to about 90 amino acid substitutions, about 60 amino acid substitutions to about 85 amino acid substitutions, about 60 amino acid substitutions to about 80 amino acid substitutions, about 60 amino acid substitutions to about 75 amino acid substitutions, about 60 amino acid substitutions to about 70 amino acid substitutions, about 60 amino acid substitutions to about 65 amino acid substitutions, between about 70 amino acid substitutions to about 160 amino acid substitutions, about 70 amino acid substitutions to about 155 amino acid substitutions
  • amino acid substitutions to about 145 amino acid substitutions about 80 amino acid substitutions to about 140 amino acid substitutions, about 80 amino acid substitutions to about 135 amino acid substitutions, about 80 amino acid substitutions to about 130 amino acid substitutions, about 80 amino acid substitutions to about 125 amino acid substitutions, about 80 amino acid substitutions to about 120 amino acid substitutions, about 80 amino acid substitutions to about 115 amino acid substitutions, about 80 amino acid substitutions to about 110 amino acid substitutions, about 80 amino acid substitutions to about 105 amino acid substitutions, about 80 amino acid substitutions to about 100 amino acid substitutions, about 80 amino acid substitutions to about 95 amino acid substitutions, about 80 amino acid substitutions to about 90 amino acid substitutions, about 80 amino acid substitutions to about 85 amino acid substitutions, between about 90 amino acid substitutions to about 160 amino acid substitutions, about 90 amino acid substitutions to about 155 amino acid substitutions, about 90 amino acid substitutions to about 150 amino acid substitutions, about 90 amino acid substitutions to about 145 amino acid substitutions, about 90 amino acid substitutions to about 140
  • substitutions to about 115 amino acid substitutions about 100 amino acid substitutions to about 110 amino acid substitutions, about 100 amino acid substitutions to about 105 amino acid substitutions, between about 110 amino acid substitutions to about 160 amino acid substitutions, about 110 amino acid substitutions to about 155 amino acid substitutions, about 110 amino acid substitutions to about 150 amino acid substitutions, about 110 amino acid substitutions to about 145 amino acid substitutions, about 110 amino acid substitutions to about 140 amino acid substitutions, about 110 amino acid substitutions to about 135 amino acid substitutions, about 110 amino acid substitutions to about 130 amino acid substitutions, about 110 amino acid substitutions to about 125 amino acid substitutions, about 110 amino acid substitutions to about 120 amino acid substitutions, about 110 amino acid substitutions to about 115 amino acid substitutions, between about 120 amino acid substitutions to about 160 amino acid substitutions, about 120 amino acid substitutions to about 155 amino acid substitutions, about 120 amino acid substitutions to about 150 amino acid substitutions, about 120 amino acid substitutions to about 145 amino acid substitutions, about 120 amino acid substitutions to about 140 amino
  • An active target protein (e.g., a supporting cell target protein or a hair cell target protein) can include, e.g., a sequence of a wildtype, lull-length target protein (e.g., a supporting cell target protein (e.g., a wildtype, human, lull-length supporting cell target protein) or a hair cell target protein (e.g., a wildtype, human, lull-length hair cell target protein) that has 1 amino acid to about 50 amino acids, 1 amino acid to about 45 amino acids, 1 amino acid to about 40 amino acids, 1 amino acid to about 35 amino acids, 1 amino acid to about 30 amino acids, 1 amino acid to about 25 amino acids, 1 amino acid to about 20 amino acids, 1 amino acid to about 15 amino acids, 1 amino acid to about 10 amino acids, 1 amino acid to about 9 amino acids, 1 amino acid to about 8 amino acids, 1 amino acid to about 7 amino acids, 1 amino acid to about 6 amino acids, 1 amino acid to about 5 amino acids, 1 amino acid to about 4 amino acids,
  • amino acids about 10 amino acids to about 20 amino acids, about 10 amino acids to about 15 amino acids, about 15 amino acids to about 50 amino acids, about 15 amino acids to about 45 amino acids, about 15 amino acids to about 40 amino acids, about 15 amino acids to about 35 amino acids, about 15 amino acids to about 30 amino acids, about 15 amino acids to about 25 amino acids, about 15 amino acids to about 20 amino acids, about 20 amino acids to about 50 amino acids, about 20 amino acids to about 45 amino acids, about 20 amino acids to about 40 amino acids, about 20 amino acids to about 35 amino acids, about 20 amino acids to about 30 amino acids, about 20 amino acids to about 25 amino acids, about 25 amino acids to about 50 amino acids, about 25 amino acids to about 45 amino acids, about 25 amino acids to about 40 amino acids, about 25 amino acids to about 35 amino acids, about 25 amino acids to about 30 amino acids, about 30 amino acids to about 50 amino acids, about 30 amino acids to about 45 amino acids, about 30 amino acids to about 40 amino acids, about 30 amino acids to about 35 amino acids, about 35 amino acids to about 30 amino acids, about 30
  • the two or more deleted amino acids can be contiguous in the sequence of the wildtype, lull-length protein. In other examples where two or more amino acids are deleted from the sequence of a wildtype, lull-length hair cell target protein, the two or more deleted amino acids are not contiguous in the sequence of the wildtype, lull-length protein.
  • amino acids that are not conserved between wildtype, lull-length hair cell target proteins from different species can be deleted without losing activity, while those amino acids that are conserved between wildtype, lull-length hair cell target proteins from different species should not be deleted as they are more likely (than amino acids that are not conserved between different species) to be involved in activity.
  • the two or more deleted amino acids can be contiguous in the sequence of the wildtype, lull-length protein.
  • two or more amino acids are deleted from the sequence of a wildtype, lull-length supporting cell target protein
  • the two or more deleted amino acids are not contiguous in the sequence of the wildtype, lull-length protein.
  • amino acids that are not conserved between wildtype, lull- length supporting cell target proteins from different species can be deleted without losing activity, while those amino acids that are conserved between wildtype, lull-length supporting cell target proteins from different species should not be deleted as they are more likely (than amino acids that are not conserved between different species) to be involved in activity.
  • an active target protein e.g., a supporting cell target protein or a hair cell target protein
  • amino acid to about 15 amino acids 1 amino acid to about 10 amino acids, 1 amino acid to about 9 amino acids, 1 amino acid to about 8 amino acids, 1 amino acid to about 7 amino acids, 1 amino acid to about 6 amino acids, 1 amino acid to about 5 amino acids, 1 amino acid to about 4 amino acids, 1 amino acid to about 3 amino acids, about 2 amino acids to about 100 amino acids, about 2 amino acid to about 95 amino acids, about 2 amino acids to about 90 amino acids, about 2 amino acids to about 85 amino acids, about
  • 2 amino acids to about 80 amino acids about 2 amino acids to about 75 amino acids, about 2 amino acids to about 70 amino acids, about 2 amino acids to about 65 amino acids, about 2 amino acids to about 60 amino acids, about 2 amino acids to about 55 amino acids, about 2 amino acids to about 50 amino acids, about 2 amino acids to about 45 amino acids, about 2 amino acids to about 40 amino acids, about 2 amino acids to
  • amino acids to about 30 amino acids about 2 amino acids to about 25 amino acids, about 2 amino acids to about 20 amino acids, about 2 amino acids to about 15 amino acids, about 2 amino acids to about 10 amino acids, about 2 amino acids to about 9 amino acids, about 2 amino acids to about 8 amino acids, about 2 amino acids to about 7 amino acids, about 2 amino acids to about 6 amino acids, about 2 amino acids to about 5 amino acids, about 2 amino acids to about 4 amino acids, about 3 amino acids to about 100 amino acids, about 3 amino acid to about 95 amino acids, about 3 amino acids to about 90 amino acids, about 3 amino acids to about 85 amino acids, about 3 amino acids to about 80 amino acids, about 3 amino acids to about 75 amino acids, about 3 amino acids to about 70 amino acids, about 3 amino acids to about 65 amino acids, about 3 amino acids to about 60 amino acids, about 3 amino acids to about 55 amino acids, about 3 amino acids to about 50 amino acids, about 3 amino acids to about 45 amino acids, about 3 amino acids to about 40 amino acids, about 3 amino acids to about 35 amino acids,
  • amino acids to about 5 amino acids about 4 amino acids to about 100 amino acids, about 4 amino acid to about 95 amino acids, about 4 amino acids to about 90 amino acids, about 4 amino acids to about 85 amino acids, about 4 amino acids to about 80 amino acids, about 4 amino acids to about 75 amino acids, about 4 amino acids to about 70 amino acids, about 4 amino acids to about 65 amino acids, about 4 amino acids to about 60 amino acids, about 4 amino acids to about 55 amino acids, about 4 amino acids to about 50 amino acids, about 4 amino acids to about 45 amino acids, about 4 amino acids to about 40 amino acids, about 4 amino acids to about 35 amino acids, about 4 amino acids to about 30 amino acids, about 4 amino acids to about 25 amino acids, about
  • 4 amino acids to about 20 amino acids about 4 amino acids to about 15 amino acids, about 4 amino acids to about 10 amino acids, about 4 amino acids to about 9 amino acids, about 4 amino acids to about 8 amino acids, about 4 amino acids to about 7 amino acids, about 4 amino acids to about 6 amino acids, about 5 amino acids to about 100 amino acids
  • amino acids about 5 amino acid to about 95 amino acids, about 5 amino acids to about 90 amino acids, about 5 amino acids to about 85 amino acids, about 5 amino acids to about 80 amino acids, about 5 amino acids to about 75 amino acids, about 5 amino acids to about 70 amino acids, about 5 amino acids to about 65 amino acids, about 5 amino acids to about 60 amino acids, about 5 amino acids to about 55 amino acids, about 5 amino acids to about 50 amino acids, about 5 amino acids to about 45 amino acids, about 5 amino acids to about 40 amino acids, about 5 amino acids to about 35 amino acids, about
  • 5 amino acids to about 30 amino acids about 5 amino acids to about 25 amino acids, about 5 amino acids to about 20 amino acids, about 5 amino acids to about 15 amino acids, about 5 amino acids to about 10 amino acids, about 5 amino acids to about 9 amino acids, about 5 amino acids to about 8 amino acids, about 5 amino acids to about 7 amino acids, about 6 amino acids to about 100 amino acids, about 6 amino acid to about 95 amino acids, about 6 amino acids to about 90 amino acids, about 6 amino acids to about 85 amino acids, about 6 amino acids to about 80 amino acids, about 6 amino acids to about 75 amino acids, about 6 amino acids to about 70 amino acids, about 6 amino acids to about 65 amino acids, about 6 amino acids to about 60 amino acids, about 6 amino acids to about 55 amino acids, about 6 amino acids to about 50 amino acids, about 6 amino acids to about 45 amino acids, about 6 amino acids to about 40 amino acids, about 6 amino acids to about 100 amino acids, about 6 amino acid to about 95 amino acids, about 6 amino acids to about 90 amino acids, about 6 amino acids to about 85 amino acids, about 6 amino acids to about
  • amino acids to about 95 amino acids about 30 amino acids to about 90 amino acids, about 30 amino acids to about 85 amino acids, about 30 amino acids to about 80 amino acids, about 30 amino acids to about 75 amino acids, about 30 amino acids to about 70 amino acids, about 30 amino acids to about 65 amino acids, about 30 amino acids to about 60 amino acids, about 30 amino acids to about 55 amino acids, about 30 amino acids to about 50 amino acids, about 30 amino acids to about 45 amino acids, about 30 amino acids to about 40 amino acids, about 30 amino acids to about 35 amino acids, about 40 amino acids to about 100 amino acids, about 40 amino acid to about 95 amino acids, about 40 amino acids to about 90 amino acids, about 40 amino acids to about 85 amino acids, about 40 amino acids to about 80 amino acids, about 40 amino acids to about 75 amino acids, about 40 amino acids to about 70 amino acids, about 40 amino acids to about 65 amino acids, about 40 amino acids to about 60 amino acids, about 40 amino acids to about 55 amino acids, about 40 amino acids to about 50 amino acids, about 40 amino acids to about 45 amino acids, about 50 amino acids to about 100
  • an active target protein e.g., a supporting cell target protein or a hair cell target protein
  • the 1 amino acid to 50 amino acids can be inserted as a contiguous sequence into the sequence of a wildtype, full-length protein. In some examples, the 1 amino acid to 50 amino acids (or any subrange thereof) are not inserted as a contiguous sequence into the sequence of a wildtype, full-length protein. As can be appreciated in the art, the 1 amino acid to 50 amino acids can be inserted into a portion of the sequence of a wildtype, full- length protein that is not well-conserved between species.
  • Figure 1A is a representative image of Myo7a/Iba-l immunofluorescent staining of cochlear tissue of a cynomolgus macaque (non-human primate) following administration of a single Anc80-GFP AAV vector directly into the inner ear through the round window.
  • Figure IB is a representative image of Anc80-GFP immunofluorescent staining of the same cochlear tissue of the cynomolgus macaque as in Figure 1 A.
  • Figure 1C is a representative image of a merged immunofluorescent staining of Myo7a/Iba-l and Anc80-GFP of the same cochlear tissue of the cynomolgus macaque as in Figure 1 A.
  • compositions that include at least two different nucleic acid vectors, where: each of the at least two different adeno-associated virus (AAV) vectors includes a coding sequence that encodes a different portion of a supporting cell target protein, each of the encoded portions being at least 30 amino acid residues in length, where the amino acid sequence of each of the encoded portions may optionally partially overlap with the amino acid sequence of a different one of the encoded portions; no single vector of the at least two different vectors encodes the lull-length supporting cell target protein; at least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of the supporting cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons; and when introduced into a primate cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a lull-length supporting cell target protein.
  • AAV
  • compositions including two different nucleic acid vectors, where: a first nucleic acid vector of the two different nucleic acid vectors includes a promoter, a first coding sequence that encodes an N-terminal portion of a supporting cell target protein positioned 3’ of the promoter, and a splicing donor signal sequence positioned at the 3’ end of the first coding sequence; and a second nucleic acid vector of the two different nucleic acid vectors includes a splicing acceptor signal sequence, a second coding sequence that encodes a C-terminal portion of a supporting cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the amino acid sequences of the encoded portions do not overlap, where no single vector of the two different vectors encodes the lull-length supporting cell target protein, and, when the coding sequence
  • compositions including: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a supporting cell target protein positioned 3’ of the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a first detectable marker gene positioned 3’ of the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second detectable marker gene, a splicing acceptor signal sequence positioned 3’ of the second detectable marker gene, a second coding sequence that encodes a C-terminal portion of a supporting cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence positioned at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the respective amino acid sequences of the encoded portions do not overlap with each other, where no single vector
  • compositions including: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a supporting cell target protein positioned 3’ to the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a FI phage recombinogenic region positioned 3’ to the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second FI phage recombinogenic region, a splicing acceptor signal sequence positioned 3’ of the second FI phage recombinogenic region, a second coding sequence that encodes a C-terminal portion of a supporting cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence positioned at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the respective
  • splicing occurs between the splicing donor signal one transcript and the splicing acceptor signal on the other transcript, thereby forming a recombined RNA molecule that encodes a lull-length supporting cell target protein.
  • compositions including a single adeno-associated virus (AAV) vector, where the single AAV vector that includes a nucleic acid sequence that encodes a supporting cell target protein; and when introduced into a primate cell, a nucleic acid encoding a lull-length supporting cell target protein is generated at the locus of the supporting cell target gene, and the primate expresses the supporting cell target protein.
  • AAV adeno-associated virus
  • compositions that include at least two different nucleic acid vectors, where: each of the at least two different adeno-associated virus (AAV) vectors includes a coding sequence that encodes a different portion of a hair cell target protein, each of the encoded portions being at least 30 amino acid residues in length, where the amino acid sequence of each of the encoded portions may optionally partially overlap with the amino acid sequence of a different one of the encoded portions; no single vector of the at least two different vectors encodes the lull-length hair cell target protein; at least one of the coding sequences includes a nucleotide sequence spanning two neighboring exons of the hair cell target genomic DNA, and lacks an intronic sequence between the two neighboring exons; and when introduced into a primate cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a lull-length hair cell target protein.
  • AAV
  • compositions including two different nucleic acid vectors, where: a first nucleic acid vector of the two different nucleic acid vectors includes a promoter, a first coding sequence that encodes an N-terminal portion of a hair cell target protein positioned 3’ of the promoter, and a splicing donor signal sequence positioned at the 3’ end of the first coding sequence; and a second nucleic acid vector of the two different nucleic acid vectors includes a splicing acceptor signal sequence, a second coding sequence that encodes a C-terminal portion of a hair cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence at
  • each of the encoded portions is at least 30 amino acid residues in length, where the amino acid sequences of the encoded portions do not overlap, where no single vector of the two different vectors encodes the lull-length hair cell target protein, and, when the coding sequences are transcribed in a primate cell, to produce RNA transcripts, splicing occurs between the splicing donor signal sequence on one transcript and the splicing acceptor signal sequence on the other transcript, thereby forming a recombined RNA molecule that encodes a lull-length hair cell target protein.
  • compositions including: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a hair cell target protein positioned 3’ of the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a first detectable marker gene positioned 3’ of the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second detectable marker gene, a splicing acceptor signal sequence positioned 3’ of the second detectable marker gene, a second coding sequence that encodes a C-terminal portion of a hair cell target protein positioned at the 3’ end of the splicing acceptor signal sequence, and a polyadenylation sequence positioned at the 3’ end of the second coding sequence; where each of the encoded portions is at least 30 amino acid residues in length, where the respective amino acid sequences of the encoded portions do not overlap with each other, where no single vector
  • compositions including: a first nucleic acid vector including a promoter, a first coding sequence that encodes an N-terminal portion of a hair cell target protein positioned 3’ to the promoter, a splicing donor signal sequence positioned at the 3’ end of the first coding sequence, and a FI phage recombinogenic region positioned 3’ to the splicing donor signal sequence; and a second nucleic acid vector, different from the first nucleic acid vector, including a second FI phage recombinogenic region, a splicing
  • each of the encoded portions is at least 30 amino acid residues in length, where the respective amino acid sequences of the encoded portions do not overlap with each other, where no single vector of the two different vectors encodes the lull-length hair cell target protein, and, when the coding sequences are transcribed in a primate cell to produce RNA transcripts, splicing occurs between the splicing donor signal one transcript and the splicing acceptor signal on the other transcript, thereby forming a recombined RNA molecule that encodes a lull-length hair cell target protein.
  • compositions including a single adeno-associated virus (AAV) vector, where the single AAV vector that includes a nucleic acid sequence that encodes a hair cell target protein; and when introduced into a primate cell, a nucleic acid encoding a lull-length hair cell target protein is generated at the locus of the hair cell target gene, and the primate expresses the hair cell target protein.
  • AAV adeno-associated virus
  • kits that include any of the compositions described herein.
  • Also provided are methods of correcting a supporting cell target gene defect in a supporting cell of an inner ear in a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering repairs the supporting cell target gene defect in the supporting cell of the inner ear of the primate.
  • Also provided are methods of correcting a hair cell target gene defect in a hair cell of an inner ear in a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering repairs the hair cell target gene defect in the hair cell of the inner ear of the primate.
  • Also provided herein are methods of increasing the expression level of a supporting cell target protein in a supporting cell of an inner ear of a primate that include:
  • administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering results in an increase in the expression level of the supporting cell target protein in the supporting cell of the inner ear of the primate.
  • Also provided herein are methods of increasing the expression level of a hair cell target protein in a hair cell of an inner ear of a primate that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein, where the administering results in an increase in the expression level of the hair cell target protein in the hair cell of the inner ear of the primate.
  • Also provided herein are methods of restoring synapses and/or preserving spiral ganglion nerves in a primate identified or diagnosed as having an inner ear disorder that include: administering to the inner ear of the primate a therapeutically effective amount of any of the compositions described herein.
  • Gap junction protein beta 2 (GJB2)
  • the GJB2 gene encodes gap junction protein beta 2, a member of the gap junction protein family. Gap junction proteins are specialized proteins, also called connexins, involved in intracellular communication. Mutations in GJB2 have been associated with hearing loss and deafness (Amorini et al., Ann. Hum. Genet. 79(5):341-349, 2015; Qing et al., Genet. Test Mol. Biomarkers 19(l):52-58, 2015).
  • the human GJB2 gene is located on chromosome 13ql2. It contains 2 exons encompassing ⁇ 5 kilobases (kb) (NCBI Accession No. NG008358.1).
  • the lull-length wildtype GJB2 protein expressed from the human GJB2 gene is 226 amino acids in length.
  • Various mutations in the GJB2 gene have been associated with hearing loss (e.g., non-syndromic sensorineural hearing loss or syndromic sensorineural hearing loss).
  • hearing loss e.g., non-syndromic sensorineural hearing loss or syndromic sensorineural hearing loss.
  • the c.35delG allele was found on 65.5% of patients from Eastern Sicily (Amorini et al., Ann. Hum. Genet. 79(5):341-349, 2015).
  • Additional exemplary mutations in a GJB2 gene detected in subjects having non- syndromic sensorineural hearing loss or syndromic sensorineural hearing loss, and methods of sequencing a nucleic acid encoding GJB2 are described in, e.g., Snoeckx et al., Am. J. Hum. Genet 77: 945-957, 2005; Welch et al., Am. J. Med. Genet A 143: 1567- 1573, 2007; and Zelante et al., Hum. Mol. Genet. 6:1605-1609, 1997.
  • Methods of detecting mutations in a gene are well-known in the art. Non-limiting examples of such techniques include: real-time polymerase chain reaction (RT-PCR), PCR, sequencing, Southern blotting, and Northern blotting.
  • An exemplary human wildtype GJB2 protein is or includes the sequence of SEQ ID NO: 1.
  • Non-limiting examples of nucleic acid encoding a wildtype GJB2 protein are or include SEQ ID NO: 2.
  • SEQ ID NO: 2 At least some or all of the codons in SEQ ID NO: 2 can be codon-optimized to allow for optimal expression in a non-human mammal.
  • the GJB2 protein comprises a sequence that is at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) identical to SEQ ID NO: 1.
  • a non-limiting example of a human wildtype GJB2 genomic DNA sequence is SEQ ID NO: 3.
  • the exons in SEQ ID NO: 3 are: nucleotide positions 1-156 (exon 1), and nucleotide positions 3336-5469 (exon 2).
  • the intron is located between each of these exons in SEQ ID NO: 3, i.e., at nucleotide positions 157-3335 (intron 1).
  • Solute carrier family 26 member 4 (SLC26A4)
  • the SLC26A4 gene encodes the Pendred protein, and has homology to sulfate transporters. Mutations in SLC26A4 have been associated with hearing loss and deafness (Albert et al., Eur. J. Hum. Genet. 14:773-779, 2006; and Qing et al., Genet. Test Mol. Biomarkers 19(l):52-58, 2015).
  • the human SLC26A4 gene is located on chromosome 7q22. It contains 21 exons encompassing ⁇ 57 kilobases (kb) (NCBI Accession No. NG 008489.1). The full-length wildtype SLC26A4 protein expressed from the human SLC26A4 gene is 780 amino acids in length.
  • Additional exemplary mutations in a SLC26A4 gene detected in subjects having non-syndromic sensorineural hearing loss or syndromic sensorineural hearing loss, and methods of sequencing a nucleic acid encoding SLC26A4 are described in, e.g., Albert et al., Eur. J. Hum. Genet. 14: 773-779, 2006; and Qing et al., Genet. Test Mol. Biomarkers 19(l):52-58, 2015. Methods of detecting mutations in a gene are well-known in the art. Non-limiting examples of such techniques include: real-time polymerase chain reaction (RT-PCR), PCR, sequencing, Southern blotting, and Northern blotting.
  • RT-PCR real-time polymerase chain reaction
  • An exemplary human wildtype SLC26A4 protein is or includes the sequence of SEQ ID NO: 4.
  • Non-limiting examples of nucleic acid encoding a wildtype SLC26A4 protein are or include SEQ ID NO: 5 or 6.
  • at least some or all of the codons in SEQ ID NO: 5 or 6 can be codon-optimized to allow for optimal expression in a non-human mammal.
  • the SLC26A4 protein comprises a sequence that is at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%) identical to SEQ ID NO: 4.
  • SEQ ID NO: 7 A non-limiting example of a human wildtype SLC26A4 genomic is SEQ ID NO: 7.
  • the exons in SEQ ID NO: 7 are: nucleotide positions 1-221 (exon 1), nucleotide positions 1005-1171 (exon 2), nucleotide positions 2262-2801 (exon 3), nucleotide positions 11504-11614 (exon 4), nucleotide positions 13530-13714 (exon 5), nucleotide positions 14311-14475 (exon 6), nucleotide positions 22568-22720 (exon 7), nucleotide positions 22821-22903 (exon 8), nucleotide positions 28419-28566 (exon 9), nucleotide positions 29490-29603 (exon 10), nucleotide positions 33769-33846 (exon 11), nucleotide positions 33987-34082 (exon 12), nucleotide positions 35299-35405 (ex
  • the introns are located between each of these exons in SEQ ID NO: 7, i.e., at nucleotide positions 222-1004 (intron 1), nucleotide positions 1172-2261 (intron 2), nucleotide positions 2802-11503 (intron 3), nucleotide positions 11615-13529 (intron 4), nucleotide positions 13715-14310 (intron 5), nucleotide positions 14476-22567 (intron 6), nucleotide positions 22721-22820 (intron 7), nucleotide positions 22904-28418 (intron 8), nucleotide positions 28567-29489 (intron 9), nucleotide positions 29604-33768 (intron 10), nucleotide positions 33847-33986 (intron 11), nucleotide positions 34083-35298 (intron 12), nucleotide positions 35406- 37407 (intron 13), nucleotide positions 37478-39448
  • gagttaatgg gtgcagtaca ccaacatggc acacgtatac atatgtaaca aacctgcacg
  • gagatctctc atgaaatgaa gagtagtcaa gacaccctac accataattt

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CN201980082439.4A CN113853437A (zh) 2018-11-07 2019-11-07 腺相关病毒载体用于在内耳中的毛细胞和支持细胞中校正基因缺陷/表达蛋白质的用途
US17/291,931 US20230056182A1 (en) 2018-11-07 2019-11-07 Use of adeno-associated viral vectors to correct gene defects/ express proteins in hair cells and supporting cells in the inner ear
EP22155160.9A EP4063510A1 (en) 2018-11-07 2019-11-07 Use of adeno-associated viral vectors to correct gene defects/ express proteins in hair cells and supporting cells in the inner ear
JP2021524060A JP2022506613A (ja) 2018-11-07 2019-11-07 内耳の有毛細胞及び支持細胞における遺伝子欠陥／発現タンパク質を修正するためのアデノ随伴ウイルスベクターの使用
EP19881503.7A EP3880826A4 (en) 2018-11-07 2019-11-07 USE OF ADENO-ASSOCIATED VIRAL VECTORS TO CORRECT GENE DEFECTS/EXPRESS PROTEINS IN CILIATE AND SUPPORTING CELLS IN THE INNER EAR
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Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021231808A3 (en) * 2020-05-13 2021-12-23 Akouos, Inc. Compositions and methods for treating gjb2-associated hearing loss
WO2022056444A1 (en) * 2020-09-14 2022-03-17 President And Fellows Of Harvard College Recombinant adeno associated virus (raav) encoding gjb2 and uses thereof
WO2022056440A1 (en) * 2020-09-14 2022-03-17 President And Fellows Of Harvard College Dual-aav vector delivery of pcdh15 and uses thereof
WO2022099007A1 (en) * 2020-11-06 2022-05-12 Applied Genetic Technologies Corporation Variant adeno-associated virus (aav)capsid polypeptides and gene therapeutics thereof for treatment of hearing loss
WO2022119839A1 (en) * 2020-12-01 2022-06-09 Akouos, Inc. Anti-vegf antibody constructs and related methods for treating vestibular schwannoma associated symptoms
WO2022146839A1 (en) 2020-12-29 2022-07-07 Akouos, Inc. Compositions and methods for treating clrn1-associated hearing loss and/or vision loss
US20220267795A1 (en) * 2021-02-24 2022-08-25 Kinase Pharma Inc. Compositions and methods for regulating production of an angiogensis inhibitor
US11525139B2 (en) 2016-08-23 2022-12-13 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US11660353B2 (en) 2018-04-27 2023-05-30 Decibel Therapeutics, Inc. Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems
US11697801B2 (en) 2017-12-19 2023-07-11 Akouos, Inc. AAV-mediated delivery of therapeutic antibodies to the inner ear
WO2023150142A1 (en) * 2022-02-02 2023-08-10 Akouos, Inc. Anti-vegf antibody constructs and related methods for treating vestibular schwannoma associated symptoms
US11807867B2 (en) 2020-02-21 2023-11-07 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
WO2024073635A3 (en) * 2022-09-30 2024-06-13 Decibel Therapeutics, Inc. Slc26a4 regulatory elements and uses thereof
US12054724B2 (en) 2018-04-10 2024-08-06 President And Fellows Of Harvard College AAV vectors encoding clarin-1 or GJB2 and uses thereof
US12129287B2 (en) 2020-09-14 2024-10-29 President And Fellows Of Harvard College Recombinant adeno associated virus encoding clarin-1 and uses thereof
US12233136B2 (en) 2018-04-27 2025-02-25 Decibel Therapeutics, Inc. Myosin 15 promoters and uses thereof
US12252520B2 (en) 2019-02-08 2025-03-18 Decibel Therapeutics, Inc. Myosin 15 promoters and uses thereof
US12410442B2 (en) 2019-02-08 2025-09-09 Regeneron Pharmaceuticals, Inc. Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems
US12589168B2 (en) 2019-10-30 2026-03-31 Regeneron Pharmaceuticals, Inc. Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems
US12612645B2 (en) 2020-04-24 2026-04-28 President And Fellows Of Harvard College AAV vectors encoding mini-PCDH15 and uses thereof

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2025179016A1 (en) * 2024-02-20 2025-08-28 Skylark Bio, Inc. Recombinant adeno associated virus and uses thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130095071A1 (en) * 2009-12-21 2013-04-18 Audigen Inc. Method of treating or preventing hearing loss
US20160076054A1 (en) * 2013-04-18 2016-03-17 Fondazione Telethon Effective delivery of large genes by dual aav vectors
WO2017136764A1 (en) * 2016-02-05 2017-08-10 The General Hospital Corporation Hybrid system for efficient gene delivery to cells of the inner ear
WO2018039375A1 (en) * 2016-08-23 2018-03-01 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0928202A4 (en) * 1996-09-25 2001-09-05 Cell Genesys Inc NON-INVASIVE ADMINISTRATION OF ADENO-ASSOCIATED VIRAL VECTORS
EP1082445A2 (en) * 1998-05-27 2001-03-14 Cell Genesys, Inc. Adeno-associated viral vector-mediated expression of factor viii activity
CA2890061A1 (en) * 2012-12-04 2014-06-12 Merck Sharp & Dohme Corp. Conditional replicating viral vectors
US9790490B2 (en) * 2015-06-18 2017-10-17 The Broad Institute Inc. CRISPR enzymes and systems
TWI813532B (zh) * 2015-06-18 2023-09-01 美商博得學院股份有限公司 降低脱靶效應的ｃｒｉｓｐｒ酶突變
SG11201804814YA (en) * 2015-12-11 2018-07-30 Massachusetts Eye & Ear Infirmary Materials and methods for delivering nucleic acids to cochlear and vestibular cells
WO2017181021A1 (en) * 2016-04-15 2017-10-19 Regenxbio Inc. Treatment of ocular diseases with fully-human post-translationally modified anti-vegf fab
KR102746901B1 (ko) * 2017-03-03 2024-12-26 옵시디안 테라퓨틱스, 인크. Cd19 조성물 및 면역요법을 위한 방법

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20130095071A1 (en) * 2009-12-21 2013-04-18 Audigen Inc. Method of treating or preventing hearing loss
US20160076054A1 (en) * 2013-04-18 2016-03-17 Fondazione Telethon Effective delivery of large genes by dual aav vectors
WO2017136764A1 (en) * 2016-02-05 2017-08-10 The General Hospital Corporation Hybrid system for efficient gene delivery to cells of the inner ear
WO2018039375A1 (en) * 2016-08-23 2018-03-01 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
ALEMI, AURASH: "AOS research grant: Restoration of Hearing in the Otoferliu Knockout Mouse using Viral Gene Therapy", 145TH ANNUAL MEETING, 21-22 APRIL 2012; MANCHESTER GRAND HYATT, SAN DIEGO, US, USA, AMERICAN OTOLOGICAL SOCIETY, INC., US, vol. 145th, 21 April 2012 (2012-04-21) - 22 April 2012 (2012-04-22), US, pages 68, XP009513930 *
MIN YONG LEE, YONG-HO PARK: "Potential of Gene and Cell Therapy for Inner Ear Hair Cells", BIOMED RESEARCH INTERNATIONAL, HINDAWI PUBLISHING CORPORATION, vol. 2018, 13 June 2018 (2018-06-13), pages 1 - 11, XP055696904, ISSN: 2314-6133, DOI: 10.1155/2018/8137614 *
See also references of EP3880826A4 *
WENJUAN ZHANG, ET AL: "Cochlear Gene Therapy for Sensorineural Hearing Loss: Current Status and Major Remaining Hurdles for Translational Success", FRONTIERS IN MOLECULAR NEUROSCIENCE, vol. 11, pages 1 - 15, XP055696898, DOI: 10.3389/fnmol.2018.00221 *

Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12071627B2 (en) 2016-08-23 2024-08-27 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US11993777B2 (en) 2016-08-23 2024-05-28 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US11781145B2 (en) 2016-08-23 2023-10-10 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US12264317B2 (en) 2016-08-23 2025-04-01 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US11525139B2 (en) 2016-08-23 2022-12-13 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US11697801B2 (en) 2017-12-19 2023-07-11 Akouos, Inc. AAV-mediated delivery of therapeutic antibodies to the inner ear
US12275960B2 (en) 2017-12-19 2025-04-15 Akouos, Inc. AAV-mediated delivery of therapeutic antibodies to the inner ear
US12077783B2 (en) 2017-12-19 2024-09-03 Akouos, Inc. AAV-mediated delivery of antibodies to the inner ear
US12054724B2 (en) 2018-04-10 2024-08-06 President And Fellows Of Harvard College AAV vectors encoding clarin-1 or GJB2 and uses thereof
US11660353B2 (en) 2018-04-27 2023-05-30 Decibel Therapeutics, Inc. Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems
US12233136B2 (en) 2018-04-27 2025-02-25 Decibel Therapeutics, Inc. Myosin 15 promoters and uses thereof
US12252520B2 (en) 2019-02-08 2025-03-18 Decibel Therapeutics, Inc. Myosin 15 promoters and uses thereof
US12410442B2 (en) 2019-02-08 2025-09-09 Regeneron Pharmaceuticals, Inc. Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems
US12589168B2 (en) 2019-10-30 2026-03-31 Regeneron Pharmaceuticals, Inc. Compositions and methods for treating sensorineural hearing loss using otoferlin dual vector systems
US12305191B2 (en) 2020-02-21 2025-05-20 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US11807867B2 (en) 2020-02-21 2023-11-07 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US12077773B2 (en) 2020-02-21 2024-09-03 Akouos, Inc. Compositions and methods for treating non-age-associated hearing impairment in a human subject
US12612645B2 (en) 2020-04-24 2026-04-28 President And Fellows Of Harvard College AAV vectors encoding mini-PCDH15 and uses thereof
WO2021231808A3 (en) * 2020-05-13 2021-12-23 Akouos, Inc. Compositions and methods for treating gjb2-associated hearing loss
EP4211151A4 (en) * 2020-09-14 2025-03-26 President and Fellows of Harvard College RECOMBINANT ADENO-ASSOCIATED VIRUS (RAAV) ENCODING GJB2 AND ITS USES
US12129287B2 (en) 2020-09-14 2024-10-29 President And Fellows Of Harvard College Recombinant adeno associated virus encoding clarin-1 and uses thereof
WO2022056440A1 (en) * 2020-09-14 2022-03-17 President And Fellows Of Harvard College Dual-aav vector delivery of pcdh15 and uses thereof
WO2022056444A1 (en) * 2020-09-14 2022-03-17 President And Fellows Of Harvard College Recombinant adeno associated virus (raav) encoding gjb2 and uses thereof
WO2022099007A1 (en) * 2020-11-06 2022-05-12 Applied Genetic Technologies Corporation Variant adeno-associated virus (aav)capsid polypeptides and gene therapeutics thereof for treatment of hearing loss
JP2023551533A (ja) * 2020-12-01 2023-12-08 アコーオス インコーポレイテッド 前庭神経鞘腫関連症状を治療するための抗ｖｅｇｆ抗体構築物及び関連する方法
US20230295287A1 (en) * 2020-12-01 2023-09-21 Akouos, Inc. Anti-vegf antibody constructs and related methods for treating vestibular schwannoma associated symptoms
WO2022119839A1 (en) * 2020-12-01 2022-06-09 Akouos, Inc. Anti-vegf antibody constructs and related methods for treating vestibular schwannoma associated symptoms
US12365726B2 (en) * 2020-12-01 2025-07-22 Akouos, Inc. Anti-VEGF antibody constructs
WO2022146839A1 (en) 2020-12-29 2022-07-07 Akouos, Inc. Compositions and methods for treating clrn1-associated hearing loss and/or vision loss
US20220267795A1 (en) * 2021-02-24 2022-08-25 Kinase Pharma Inc. Compositions and methods for regulating production of an angiogensis inhibitor
WO2023150142A1 (en) * 2022-02-02 2023-08-10 Akouos, Inc. Anti-vegf antibody constructs and related methods for treating vestibular schwannoma associated symptoms
WO2024073635A3 (en) * 2022-09-30 2024-06-13 Decibel Therapeutics, Inc. Slc26a4 regulatory elements and uses thereof

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JP2022506613A (ja) 2022-01-17
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