Gene Therapy Constructs and Methods for Treatment of Hearing Loss
Disclosed are compositions and methods that useful in the treatment and/or prevention of hearing loss caused by genetic mutation of the TMPRSS3 gene or the LOXHD1 gene. The compositions and methods disclosed herein use adeno-associated viral (AAV) vector gene delivery of TRMPSS3 or LOXHD1 into the inner ear to restore activity of the TMPRSS3 gene or the LOXHD1 gene, respectively, promote hair cell survival and restore hearing in patients suffering from hearing loss.
1 . An expression vector comprising:
a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:1, SEQ ID NO:3 or a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:3; and
a promoter operatively linked to the nucleic acid sequence.
2 . The expression vector of claim 1 , wherein the expression vector is an adeno-associated viral vector.
3 . The expression vector of claim 2 , wherein the adeno-associated viral vector is selected from the group consisting of AAV2, AAV2/Anc80, AAV5, AAV6, AAV6.2, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 or Anc80.
4 . The expression vector of claim 1 , wherein the promoter is selected from the group consisting of: TMPRSS3 promoters, Myo7a promoters, human cytomegalovirus (HCMV) promoters, cytomegalovirus/chicken beta-actin (CBA) promoters and Pou4f3 promoters.
5 . A pharmaceutical composition for use in a method for the treatment or prevention of hearing loss comprising an expression vector comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:1, SEQ ID NO:3 or a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:3; and a promoter operatively linked to the nucleic acid.
6 . The pharmaceutical composition of claim 5 , wherein the expression vector is an adeno-associated viral vector.
7 . The pharmaceutical composition of claim 6 , wherein the adeno-associated viral vector is selected from the group consisting of AAV2, AAV2/Anc80, AAV5, AAV6, AAV6.2, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 or Anc80.
8 . The pharmaceutical composition of claim 5 , wherein the promoter is selected from the group consisting of: TMPRSS3 promoters, Myo7a promoters, human cytomegalovirus (HCMV) promoters, cytomegalovirus/chicken beta-actin (CBA) promoters and Pou4f3 promoters.
9 . A cell transfected with an expression vector comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:1, SEQ ID NO:3 or a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:3; and a promoter operatively linked to the nucleic acid.
10 . The cell of claim 9 , wherein the expression vector is an adeno-associated viral vector.
11 . The cell of claim 10 , wherein the adeno-associated viral vector is selected from the group consisting of AAV2, AAV2/Anc80, AAV5, AAV6, AAV6.2, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 or Anc80.
12 . The cell of claim 9 , wherein the promoter is selected from the group consisting of TMPRSS3 promoters, Myo7a promoters, human cytomegalovirus (HCMV) promoters, cytomegalovirus/chicken beta-actin (CBA) promoters or Pou4f3 promoters.
13 . The cell of claim 9 , in the cell is a stem cell.
14 . The cell of claim 13 , wher the stem cell is an induced pluripotent stem cell.
15 . A method for treating or preventing hearing loss, comprising administering to a subject in need thereof an effective amount of an expression vector comprising a nucleic acid sequence selected from the group consisting of SEQ ID NO:1, a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:1, SEQ ID NO:3 or a nucleic acid sequence having at least 90% sequence identity to the nucleic acid of SEQ ID NO:3; and a promoter operatively linked to the nucleic acid.
16 . The method of claim 15 , wherein the expression vector is an adeno-associated viral vector.
17 . The method of claim 16 , wherein the adeno-associated viral vector is selected from the group consisting of AAV2, AAV2/Anc80, AAVS, AAV6, AAV6.2, AAV7, AAV8, AAV9, AAVrh8, AAVrh10, AAVrh39, AAVrh43, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8 or Anc80.
18 . The method of claim 15 , wherein the promoter is selected from the group consisting of TMPRSS3 promoters, Myo7a promoters, human cytomegalovirus (HCMV) promoters, cytomegalovirus/chicken beta-actin (CBA) promoters, or Pou4f3 promoters.
19 . The method of claim 15 , wherein the expression vector is administered by injection into the inner ear of the subject.
20 . The method of claim 19 , wherein the injection method is selected from the group consisting of cochleostomy, round window membrane, endolymphatic sac, scala media, canalostomy, scala media via the endolymphatic sac, or any combination thereof.
21 . The method of claim 15 , wherein the subject has one or more genetic risk factors associated with hearing loss.
22 . The method of claim 21 , wherein one of the genetic risk factors is selected from the group consisting of a mutation in the TMPRSS3 gene or a mutation in the LOXHD1 gene.
23 . The method of claim 21 , wherein the subject does not exhibit any clinical indicators of hearing loss.
24 . A transgenic mouse comprising a mutation that causes hearing loss selected from the group consisting of a mutation in the human LOXHD1 gene or a mutation in the human TMPRSS3 gene.