Granted Patent
B2
US 12,612,645 · App. 17/606,280 · Granted Apr 28, 2026
AAV vectors encoding mini-PCDH15 and uses thereof
Inventors:
David P. Corey (Cambridge, MA); Artur Indzhykulian (Boston, MA); Marcos Sotomayor (Columbus, OH); Maryna V. Ivanchenko (Cambridge, MA); Cole W. D. Peters (Cambridge, MA)
Assignees:
President and Fellows of Harvard College; Ohio State Innovation Foundation; Massachusetts Eye and Ear Infirmary
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Abstract
Aspects of the disclosure relate to compositions, nucleic acids, vectors, viruses, and methods useful for treating hearing loss and/or blindness, for example, Usher Syndrome type IF. The present disclosure provides isolated nucleic acids, vectors, and rAAV.9.PHP.B comprising a transgene encoding a mini-PCDH15, and methods of treating hearing loss using the same. The present disclosure also provides a gRNA associated with a base editor to correct one or more mutations in PCDH15 for treating hearing loss and/or vision loss.
Claims (15)
1 . An isolated nucleic acid comprising a transgene flanked by adeno-associated virus (AAV) inverted terminal repeats (ITRs), wherein the transgene encodes a mini-Protocadherin related 15 (mini-PCDH15) protein comprising an amino acid sequence at least 95% identical to SEQ ID NO: 95 or 99.
2 . The isolated nucleic acid of claim 1 , wherein the mini-PCDH15 protein comprises the amino acid sequence of SEQ ID NO: 95.
3 . The isolated nucleic acid of claim 2 , wherein amino acid sequence of SEQ ID NO: 95 is encoded by the nucleic acid sequence of SEQ ID NO: 97.
4 . The isolated nucleic acid of claim 1 further comprising a promoter operably linked to the transgene encoding the mini-PCDH15 protein.
5 . The isolated nucleic acid of claim 4 , wherein the promoter is a cytomegalovirus (CMV) promoter, a hybrid cytomegalovirus (CMV) immediate-early/chicken beta-actin (CAG) promoter, a chicken beta actin (CBA) promoter, or a minimal promoter.
6 . The isolated nucleic acid of claim 4 , wherein the promoter is a Methyl-CpG Binding Protein 2 (MeCP2) promoter, a Ubiquitin-C (UbiC) promoter, a Bestrophin 1(Best1) (retina native) promoter, a human red opsin (RedO) promoter, a human rhodopsin kinase (RK) promoter, a mouse cone arrestin (CAR) promoter, a human rhodopsin (Rho) promoter, a UV opsin-specific 1 (opn1sw1) promoter, a UV opsin-specific 2 (opn1sw2) promoter, an Opsin 1, Medium Wave Sensitive 2 (opn1mw2) promoter, an opsin 1, long2 wave-sensitive 1 (opn1lw1) promoter, a blue cone specific (sws2) promoter, an L-opsin (opn1lw1-cxxc1) promoter, a thyroid hormone receptor β (thrb) promoter, an LIM Homeobox 1a (Ihx1a) promoter, a connexin 55.5 (cx55.5) promoter, a metabotropic glutamate receptor 6b (grm6b) promoter, a glial fibrillar acidic protein (gfap) promoter, a cone transducin alpha subunit (gnat2) promoter, a connexin 52.7 (cx52.7) promoter, a connexin 52.9 (cx52.9) promoter, a heat shock cognate 70-kd protein,-like (hsp70l) promoter, a yeast transcription activator protein-(GAL4-VP16) promoter, an upstream activation sequence (UAS), a visual system homeobox 1 (vsx1) promoter, or a rhodopsin (zop) promoter.
7 . The isolated nucleic acid of claim 5 , wherein the minimal promoter is a minimal CMV promoter, a CMV584 bp promoter, or a JeT promoter.
8 . The isolated nucleic acid of claim 1 , wherein the AAV ITRs are AAV2 ITRs.
9 . A vector comprising the isolated nucleic acid of claim 1 .
10 . An isolated host cell comprising the isolated nucleic acid of claim 1 .
11 . A pharmaceutical composition comprising the isolated nucleic acid of claim 1 and a pharmaceutically acceptable carrier.
12 . The isolated nucleic acid of claim 1 , wherein the mini-PCDH15 protein comprises the amino acid sequence of SEQ ID NO: 99.
13 . The isolated nucleic acid of claim 12 , wherein the amino acid sequence of SEQ ID NO: 99 is encoded by the nucleic acid sequence of SEQ ID NO: 101.
14 . The isolated nucleic acid of claim 1 , wherein the mini-PCDH15 protein comprises an amino acid sequence at least 98% identical to SEQ ID NO: 95 or 99.
15 . The isolated nucleic acid of claim 1 , wherein the mini-PCDH15 protein comprises an amino acid sequence at least 99% identical to SEQ ID NO: 95 or 99.
Assignments (3)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Sep 26, 2022
From: SOTOMAYER, MARCOS
To: OHIO STATE INNOVATION FOUNDATION
Reel/Frame 061210/0755 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Sep 26, 2022
From: INDZHYKULIAN, ARTUR
To: MASSACHUSETTS EYE AND EAR INFIRMARY
Reel/Frame 061210/0861 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Sep 15, 2022
From: COREY, DAVID P.; PETERS, COLE W.D.; IVANCHENKO, MARYNA V.
To: PRESIDENT AND FELLOWS OF HARVARD COLLEGE
Reel/Frame 061099/0537 →
Continuity (2)
Provisional Application
62839543
· Apr 26, 2019
References Cited (203)
US 5399363A
· Liversidge et al.
· 1995
[cited by applicant]
US 5478745A
· Samulski et al.
· 1995
[cited by applicant]
US 5543158A
· Gref et al.
· 1996
[cited by applicant]
US 5552157A
· Yagi et al.
· 1996
[cited by applicant]
US 5565213A
· Nakamori et al.
· 1996
[cited by applicant]
US 5567434A
· Szoka, Jr.
· 1996
[cited by applicant]
US 5641515A
· Ramtoola
· 1997
[cited by applicant]
US 5738868A
· Shinkarenko
· 1998
[cited by applicant]
US 5741516A
· Webb et al.
· 1998
[cited by applicant]
US 5795587A
· Gao et al.
· 1998
[cited by applicant]
US 6001650A
· Colosi
· 1999
[cited by applicant]
US 6156303A
· Russell et al.
· 2000
[cited by applicant]
US 20030138772A1
· Gao et al.
· 2003
[cited by applicant]
US 20130095071A1
· Bance et al.
· 2013
[cited by applicant]
US 20170037479A1
· Kuznetsov et al.
· 2017
[cited by applicant]
US 20180327779A1
· Colella et al.
· 2018
[cited by applicant]
US 20240016955A1
· Corey et al.
· 2024
[cited by applicant]
WO WO98010088A1
· 1998
[cited by applicant]
WO WO2016139321A1
· 2016
[cited by applicant]
WO WO2017193059A1
· 2017
[cited by applicant]
WO WO2018031683A1
· 2018
[cited by applicant]
WO WO2018145111A1
· 2018
[cited by applicant]
WO WO2019199689A1
· 2019
[cited by applicant]
WO WO2020079034A2
· 2020
[cited by applicant]
WO WO2020097372A1
· 2020
[cited by applicant]
WO WO2020198737A1
· 2020
[cited by applicant]
Perreault-Micale, Cynthia, et al. The Journal of Molecular Diagnostics 16.6 (2014): 673-678 (Year: 2014).
[cited by examiner]
Ostedgaard LS, et. al. Proc Natl Acad Sci U S A. Feb. 22, 2005;102(8):2952-7 (Year: 2005).
[cited by examiner]
Karpati et al., The principles of gene therapy in Duchenne muscular dystrophy. Clin Invest Med. Oct. 1994;17(5):499-509.
[cited by applicant]
Invitation to Pay Additional Fees for Application No. PCT/US2020/029968, mailed Aug. 3, 2020.
[cited by applicant]
International Search Report and Written Opinion for Application No. PCT/US2020/029968, mailed Oct. 5, 2020.
[cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/US2020/029968, mailed Nov. 4, 2021.
[cited by applicant]
International Search Report and Written Opinion for Application No. PCT/US2021/050188, mailed Dec. 20, 2021.
[cited by applicant]
Ahmad et al., Restoration of connexin26 protein level in the cochlea completely rescues hearing in a mouse model of human connexin30-linked deafness. Proc Natl Acad Sci U S A. Jan. 23, 2007;104(4):1337-41. doi: 10.1073/…
[cited by applicant]
Ahmed et al., Mutations of the protocadherin gene PCDH15 cause Usher syndrome type 1F. Am J Hum Genet. Jul. 2001;69(1):25-34. doi: 10.1086/321277. Epub Jun. 7, 2001.
[cited by applicant]
Ahmed et al., PCDH15 is expressed in the neurosensory epithelium of the eye and ear and mutant alleles are responsible for both USH1F and DFNB23. Hum Mol Genet. Dec. 15, 2003;12(24):3215-23. doi: 10.1093/hmg/ddg358. Epu…
[cited by applicant]
Akil et al., Restoration of hearing in the VGLUT3 knockout mouse using virally mediated gene therapy. Neuron. Jul. 26, 2012;75(2):283-93. doi: 10.1016/j.neuron.2012.05.019.
[cited by applicant]
Alagramam et al., Mutations in protocadherin 15 and cadherin 23 affect tip links and mechanotransduction in mammalian sensory hair cells. PLoS One. Apr. 21, 2011;6(4):e19183. doi: 10.1371/journal.pone.0019183.
[cited by applicant]
Al-Moyed et al., A dual-AAV approach restores fast exocytosis and partially rescues auditory function in deaf otoferlin knock-out mice. EMBO Mol Med. Jan. 2019;11(1):e9396. doi: 10.15252/emmm.201809396.
[cited by applicant]
Angueyra et al., Leveraging Zebrafish to Study Retinal Degeneration. Front Cell Dev Biol. Sep. 19, 2018;6:110. doi: 10.3389/fcell.2018.00110. eCollection 2018.
[cited by applicant]
Araya-Secchi et al., An elastic element in the protocadherin-15 tip link of the inner ear. Nat Commun. Nov. 18, 2016;7:13458. doi: 10.1038/ncomms13458.
[cited by applicant]
Assad et al., An active motor model for adaptation by vertebrate hair cells. J Neurosci. Sep. 1992;12(9):3291-309. doi: 10.1523/JNEUROSCI.12-09-03291.1992.
[cited by applicant]
Assad et al., Tip-link integrity and mechanical transduction in vertebrate hair cells. Neuron. Dec. 1991;7(6):985-94. doi: 10.1016/0896-6273(91)90343-x.
[cited by applicant]
Azaiez et al., Genomic Landscape and Mutational Signatures of Deafness-Associated Genes. Am J Hum Genet. Oct. 4, 2018;103(4):484-497. doi: 10.1016/j.ajhg.2018.08.006. Epub Sep. 20, 2018.
[cited by applicant]
Ben-Yosef et al., A mutation of PCDH15 among Ashkenazi Jews with the type 1 Usher syndrome. N Engl J Med. Apr. 24, 2003;348(17):1664-70. doi: 10.1056/NEJMoa021502.
[cited by applicant]
Bird et al., Harnessing molecular motors for nanoscale pulldown in live cells. Mol Biol Cell. Feb. 1, 2017;28(3):463-475. doi: 10.1091/mbc.E16-08-0583. Epub Dec. 8, 2016.
[cited by applicant]
Boulay et al., Hearing is normal without connexin30. J Neurosci. Jan. 9, 2013;33(2):430-4. doi: 10.1523/JNEUROSCI.4240-12.2013.
[cited by applicant]
Brownstein et al., The R245X mutation of PCDH15 in Ashkenazi Jewish children diagnosed with nonsyndromic hearing loss foreshadows retinitis pigmentosa. Pediatr Res. Jun. 2004;55(6):995-1000. doi: 10.1203/01.PDR.00001252…
[cited by applicant]
Buenrostro et al., Transposition of native chromatin for fast and sensitive epigenomic profiling of open chromatin, DNA-binding proteins and nucleosome position. Nature Methods. Dec. 2013;10(12):1213-8. doi: 10.1038/nme…
[cited by applicant]
Burset et al., SpliceDB: database of canonical and noncanonical mammalian splice sites, Nucleic Acids Res. Jan. 1, 2001;29(1):255-9. doi: 10.1093/nar/29.1.255.
[cited by applicant]
Chamberlain et al., Progress toward Gene Therapy for Duchenne Muscular Dystrophy. Mol Ther. May 3, 2017;25(5):1125-1131. doi: 10.1016/j.ymthe.2017.02.019. Epub Apr. 15, 2017.
[cited by applicant]
Chang et al., Functional studies reveal new mechanisms for deafness caused by connexin mutations. Otol Neurotol. Feb. 2009;30(2):237-40. doi: 10.1097/MAO.0b013e318194f774.
[cited by applicant]
Chang et al., Gap junction mediated intercellular metabolite transfer in the cochlea is compromised in connexin30 null mice. PloS One. 2008;3(12):e4088. doi: 10.1371/journal.pone.0004088. Epub Dec. 31, 2008.
[cited by applicant]
Chang et al., Timed conditional null of connexin26 in mice reveals temporary requirements of connexin26 in key cochlear developmental events before the onset of hearing. Neurobiol Dis. Jan. 2015;73:418-27. doi: 10.1016/…
[cited by applicant]
Chen et al., Characterization of a knock-in mouse model of the homozygous p.V37I variant in Gjb2. Sci Rep. Sep. 13, 2016;6:33279. doi: 10.1038/srep33279.
[cited by applicant]
Chen et al., Developmental abnormalities in supporting cell phalangeal processes and cytoskeleton in the Gjb2 knockdown mouse model. Dis Model Mech. Feb. 26, 2018;11(2):dmm033019. doi: 10.1242/dmm.033019.
[cited by applicant]
Chen et al., Down regulated connexin26 at different postnatal stage displayed different types of cellular degeneration and formation of organ of Corti. Biochem Biophys Res Commun. Feb. 28, 2014;445(1):71-7. doi: 10.1016…
[cited by applicant]
Cheung et al., Ca2+ changes the force sensitivity of the hair-cell transduction channel. Biophys J. Jan. 1, 2006;90(1):124-39. doi: 10.1529/biophysj.105.061226. Epub Oct. 7, 2005.
[cited by applicant]
Christensen et al., TRP channels in mechanosensation : Direct or indirect activation? Nat Rev Neurosci. Jul. 2007;8(7):510-21. doi: 10.1038/nrn2149.
[cited by applicant]
Chylinski et al., The tracrRNA and Cas9 families of type II CRISPR-Cas immunity systems. RNA Biol. May 2013;10(5):726-37. doi: 10.4161/rna.24321. Epub Apr. 5, 2013.
[cited by applicant]
Cohen-Salmon et al., Targeted ablation of connexin26 in the inner ear epithelial gap junction network causes hearing impairment and cell death. Curr Biol. Jul. 9, 2002;12(13):1106-11. doi: 10.1016/s0960-9822(02)00904-1.
[cited by applicant]
Corey et al., Kinetics of the receptor current in bullfrog saccular hair cells. J Neurosci. May 1983;3(5):962-76. doi: 10.1523/JNEUROSCI.03-05-00962.1983.
[cited by applicant]
Corey et al., TRPA1 is a candidate for the mechanosensitive transduction channel of vertebrate hair cells. Nature. Dec. 9, 2004;432(7018):723-30. doi: 10.1038/nature03066. Epub Oct. 13, 2004.
[cited by applicant]
Creyghton et al., Histone H3K27ac separates active from poised enhancers and predicts developmental state. Proc Natl Acad Sci U S A. Dec. 14, 2010;107(50):21931-6. doi: 10.1073/pnas.1016071107. Epub Nov. 24, 2010.
[cited by applicant]
Crispino et al., BAAV mediated GJB2 gene transfer restores gap junction coupling in cochlear organotypic cultures from deaf Cx26Sox10Cre mice. PloS One. 2011;6(8):e23279. doi: 10.1371/journal.pone.0023279. Epub Aug. 18,…
[cited by applicant]
Crispino et al., In vivo genetic manipulation of inner ear connexin expression by bovine adeno-associated viral vectors. Sci Rep. Aug. 4, 2017;7(1):6567. doi: 10.1038/s41598-017-06759-y.
[cited by applicant]
Darcy et al. Dual adeno-associated viral Anc80 vector efficiently transduces inner ear cells in non-human primates. ARO Abstract 691. 2020; 43: 447-448.
[cited by applicant]
Degen et al., Connexin32 can restore hearing in connexin26 deficient mice. Eur J Cell Biol. Oct. 2011;90(10):817-24. doi: 10.1016/j.ejcb.2011.05.001. Epub Aug. 2, 2011.
[cited by applicant]
Del Castillo et al., A novel deletion involving the connexin-30 gene, del(GJB6-d13s1854), found in trans with mutations in the GJB2 gene (connexin-26) in subjects with DFNB1 non-syndromic hearing impairment. J Med Genet…
[cited by applicant]
Del Castillo et al., DFNB1 Non-syndromic Hearing Impairment: Diversity of Mutations and Associated Phenotypes. Front Mol Neurosci. Dec. 22, 2017;10:428. doi: 10.3389/fnmol.2017.00428. eCollection 2017.
[cited by applicant]
De-La-Torre et al., A Mechanically Weak Extracellular Membrane-Adjacent Domain Induces Dimerization of Protocadherin-15. Biophys J. Dec. 18, 2018;115(12):2368-2385. doi: 10.1016/j.bpj.2018.11.010. Epub Nov. 16, 2018.
[cited by applicant]
Deltcheva et al., CRISPR RNA maturation by trans-encoded small RNA and host factor 10 RNase III. Nature. Mar. 31, 2011;471(7340):602-7. doi: 10.1038/nature09886.
[cited by applicant]
Derby et al., Gene transfer into the mammalian inner ear using HSV-1 and vaccinia virus vectors. Hear Res. Aug. 1999;134(1-2):1-8. doi: 10.1016/s0378-5955(99)00045-3.
[cited by applicant]
Deverman et al., Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain. Nat Biotechnol. Feb. 2016;34(2):204-9. doi: 10.1038/nbt.3440. Epub Feb. 1, 2016.
[cited by applicant]
Dinh et al., Diverse deafness mechanisms of connexin mutations revealed by studies using in vitro approaches and mouse models. Brain Res. Jun. 24, 2009;1277:52-69. doi: 10.1016/j.brainres.2009.02.008. Epub Feb. 20, 2009.
[cited by applicant]
Doucette et al., Profound, prelingual nonsyndromic deafness maps to chromosome 10q21 and is caused by a novel missense mutation in the Usher syndrome type IF gene PCDH15. Eur J Hum Genet. May 2009;17(5):554-64. doi: 10.…
[cited by applicant]
Dyka et al., Dual adeno-associated virus vectors result in efficient in vitro and in vivo expression of an oversized gene, MYO7A. Hum Gene Ther Methods. Apr. 2014;25(2):166-77. doi: 10.1089/hgtb.2013.212.
[cited by applicant]
Ellis et al., A survey of ex vivo/in vitro transduction efficiency of mammalian primary cells and cell lines with Nine natural adeno-associated virus (AAV1-9) and one engineered adeno- associated virus serotype. Virol J…
[cited by applicant]
Feigenspan et al., Expression of connexin36 in cone pedicles and OFF-cone bipolar cells of the mouse retina. J Neurosci. Mar. 31, 2004;24(13):3325-34. doi: 10.1523/JNEUROSCI.5598-03.2004.
[cited by applicant]
Ferretti et al., Complete genome sequence of an M1 strain of
[cited by applicant]
Fetoni et al., Cx26 partial loss causes accelerated presbycusis by redox imbalance and dysregulation of Nfr2 pathway. Redox Biol. Oct. 2018;19:301-317. doi: 10.1016/j.redox.2018.08.002. Epub Aug. 7, 2018.
[cited by applicant]
Forge et al., Gap junctions and connexin expression in the inner ear. Novartis Found Symp. 1999;219:134-50; discussion 151-6. doi: 10.1002/9780470515587.ch9.
[cited by applicant]
Forge et al., Gap junctions in the inner ear: comparison of distribution patterns in different vertebrates and assessement of connexin composition in mammals. J Comp Neurol. Dec. 8, 2003;467(2):207-31. doi: 10.1002/cne.…
[cited by applicant]
Garcia et al., Localization of myosin-Iβ near both ends of tip links in frog saccular hair cells. J Neurosci. Nov. 1, 1998;18(21):8637-47. doi: 10.1523/JNEUROSCI.18-21-08637.1998.
[cited by applicant]
Gaudelli et al., Programmable base editing of A*T to G*C in genomic DNA without DNA cleavage. Nature. Nov. 23, 2017;551(7681):464-471. doi: 10.1038/nature24644. Epub Oct. 25, 2017.
[cited by applicant]
Ge et al., Structure of mouse protocadherin 15 of the stereocilia tip link in complex with LHFPL5. Elife. Aug. 2, 2018;7:e38770. doi: 10.7554/eLife.38770.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NC_000010, version NC_000010.10.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142763, version NM_001142763.2.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142763, version NM_001142763.1.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142769, version NM_001142769.2.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142769, version NM_001142769.3.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142771, version NM_001142771.1.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142771, version NM_001142771.2.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_001142772, version NM_001142772.1.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NM_033056, version NM_033056.3.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NP_001136235, version NP_001136235.1. protocadherin-15 isoform CD1-1 precursor [
[cited by applicant]
Genbank Submission. NCBI; Accession No. NP_001136241, version NP_001136241.1. protocadherin-15 isoform CD2-1 precursor [
[cited by applicant]
Genbank Submission. NCBI; Accession No. NP_001341349, version NP_001341349.1. protocadherin-15 isoform CD3-a precursor [
[cited by applicant]
Gregorevic et al., rAAV6-microdystrophin preserves muscle function and extends lifespan in severely dystrophic mice. Nat Med. Jul. 2006;12(7):787-9. doi: 10.1038/nm1439. Epub Jul. 2, 2006.
[cited by applicant]
Grimm et al., In vitro and in vivo gene therapy vector evolution via multispecies interbreeding and retargeting of adeno-associated viruses. J Virol. Jun. 2008;82(12):5887-911. doi: 10.1128/JVI.00254-08. Epub Apr. 9, 20…
[cited by applicant]
György et al., Allele-specific gene editing prevents deafness in a model of dominant progressive hearing loss. Nat Med. Jul. 2019;25(7):1123-1130. doi: 10.1038/s41591-019-0500-9. Epub Jul. 3, 2019.
[cited by applicant]
György et al., Gene transfer with AAV9-PHP.B rescues hearing in a mouse model of usher syndrome 3A and transduces hair cells in a non-human primate. Mol Ther Methods Clin Dev. Nov. 20, 2018;13:1-13. doi: 10.1016/j.omtm.…
[cited by applicant]
György et al., Rescue of hearing by gene delivery to inner-ear hair cells using exosome-associated AAV. Mol Ther. Feb. 1, 2017;25(2):379-391. doi: 10.1016/j.ymthe.2016.12.010. Epub Jan. 9, 2017.
[cited by applicant]
Hanlon et al., AAV-S: A novel AAV vector selected in brain transduces the inner ear with high efficiency. Mol Ther. Apr. 28, 2020; 18 (4S1): Abstract 151.
[cited by applicant]
Hanlon et al., Selection of an Efficient AAV Vector for Robust CNS Transgene Expression. Mol Ther Methods Clin Dev. Oct. 23, 2019;15:320-332. doi: 10.1016/j.omtm.2019.10.007. eCollection Dec. 13, 2019.
[cited by applicant]
Hasson et al., Unconventional myosins in inner-ear sensory epithelia J Cell Biol. Jun. 16, 1997;137(6):1287-307. doi: 10.1083/jcb.137.6.1287.
[cited by applicant]
Haywood-Watson et al., Ames Waltzer deaf mice have reduced electroretinogram amplitudes and complex alternative splicing of Pedh15 transcripts. Invest Ophthalmol Vis Sci. Jul. 2006;47(7):3074-84. doi: 10.1167/iovs.06-01…
[cited by applicant]
Hirsch et al., Delivering Transgenic DNA Exceeding the Carrying Capacity of AAV Vectors. Methods Mol Biol. 2016;1382:21-39. doi: 10.1007/978-1-4939-3271-9_2.
[cited by applicant]
Holt et al., A chemical-genetic strategy demonstrates myosin 1c mediates sensory adaptation in hair cells. Cell. Feb. 8, 2002;108(3):371-81. doi: 10.1016/s0092-8674(02)00629-3.
[cited by applicant]
Holt et al., Functional expression of exogenous proteins in mammalian sensory hair cells infected with adenoviral vectors. J Neurophysiol. Apr. 1999;81(4):1881-8. doi: 10.1152/jn.1999.81.4.1881.
[cited by applicant]
Hrvatin et al., Single-cell analysis of experience-dependent transcriptomic states in the mouse visual cortex. Nat Neurosci. Jan. 2018;21(1):120-129. doi: 10.1038/s41593-017-0029-5. Epub Dec. 11, 2017.
[cited by applicant]
Iizuka et al., Perinatal Gjb2 gene transfer rescues hearing in a mouse model of hereditary deafness. Hum Mol Genet. Jul. 1, 2015;24(13):3651-61. doi: 10.1093/hmg/ddv109. Epub Mar. 23, 2015.
[cited by applicant]
Indzhykulian et al., Molecular remodeling of tip links underlies mechanosensory regeneration in auditory hair cells. PLoS Biol. 2013;11(6):e1001583. doi: 10.1371/journal.pbio.1001583. Epub Jun. 11, 2013.
[cited by applicant]
Isgrig et al., AAV2.7m8 is a powerful viral vector for inner ear gene therapy. Nat Commun. Jan. 25, 2019;10(1):427. doi: 10.1038/s41467-018-08243-1.
[cited by applicant]
Ivanchenko, Preclinical testing of AAV9-PHP.B for transgene expression in the non-human primate cochlea. Hear Res. Sep. 1, 2020;394:107930. doi: 10.1016/j.heares.2020.107930. Epub Feb. 26, 2020.
[cited by applicant]
Jinek et al., A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science. Aug. 17, 2012;337(6096):816-21. doi: 10.1126/science.1225829. Epub Jun. 28, 2012.
[cited by applicant]
Johnson et al., Connexin-Mediated Signaling in Nonsensory Cells Is Crucial for the Development of Sensory Inner Hair Cells in the Mouse Cochlea. J Neurosci. Jan. 11, 2017;37(2):258-268. doi: 10.1523/JNEUROSCI.2251-16.20…
[cited by applicant]
Kamiya et al., Assembly of the cochlear gap junction macromolecular complex requires connexin 26. J Clin Invest. Apr. 2014;124(4):1598-607. doi: 10.1172/JCI67621. Epub Mar. 3, 2014.
[cited by applicant]
Kazmierczak et al., Cadherin 23 and protocadherin 15 interact to form tip-link filaments in sensory hair cells. Nature. Sep. 6, 2007;449(7158):87-91. doi: 10.1038/nature06091.
[cited by applicant]
Kelsell et al., Connexin 26 mutations in hereditary non-syndromic sensorineural deafness. Nature. May 1, 1997;387(6628):80-3. doi: 10.1038/387080a0.
[cited by applicant]
Kenna et al., Audiologic phenotype and progression in GJB2 (Connexin 26) hearing loss. Arch Otolaryngol Head Neck Surg. Jan. 2010;136(1):81-7. doi: 10.1001/archoto.2009.202.
[cited by applicant]
Kikuchi et al., Gap junctions in the rat cochlea: immunohistochemical and ultrastructural analysis. Anat Embryol (Berl) Feb. 1995;191(2):101-18. doi: 10.1007/BF00186783.
[cited by applicant]
Kleinstiver et al., Broadening the targeting range of
[cited by applicant]
Kleinstiver et al., Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature. Jul. 23, 2015;523(7561):481-5. doi: 10.1038/nature14592. Epub Jun. 22, 2015.
[cited by applicant]
Kodippili et al., Dual AAV Gene Therapy for Duchenne Muscular Dystrophy with a 7-kb Mini-Dystrophin Gene in the Canine Model. Hum Gene Ther. Mar. 2018;29(3):299-311. doi: 10.1089/hum.2017.095. Epub Aug. 4, 2017.
[cited by applicant]
Koehler et al., Generation of inner ear organoids containing functional hair cells from human pluripotent stem cells. Nat Biotechnol. Jun. 2017;35(6):583-589. doi: 10.1038/nbt.3840. Epub May 1, 2017.
[cited by applicant]
Kohrman et al., Gene therapy for deafness. Gene Ther. Dec. 2013;20(12):1119-23. doi: 10.1038/gt.2013.39. Epub Jul. 18, 2013.
[cited by applicant]
Komor et al., Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature. May 19, 2016;533(7603):420-4. doi: 10.1038/nature17946. Epub Apr. 20, 2016.
[cited by applicant]
Kwan et al., TRPA1 contributes to cold, mechanical, and chemical nociception but is not essential for hair-cell transduction. Neuron. Apr. 20, 2006;50(2):277-89. doi: 10.1016/j.neuron.2006.03.042.
[cited by applicant]
Lang et al., Effects of chronic furosemide treatment and age on cell division in the adult gerbil inner ear. J Assoc Res Otolaryngol. Jun. 2003;4(2):164-75. doi: 10.1007/s10162-002-2056-4.
[cited by applicant]
Lee et al., Mice with conditional deletion of Cx26 exhibit no vestibular phenotype despite secondary loss of Cx30 in the vestibular end organs. Hear Res. Oct. 2015;328:102-12. doi: 10.1016/j.heares.2015.07.018. Epub Jul…
[cited by applicant]
Levy et al., Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated viruses. Nat Biomed Eng. Jan. 2020;4(1):97-110. doi: 10.1038/s41551-019-0501-5. Epub Jan…
[cited by applicant]
Li et al., Characterization of slow-cycling cells in the mouse cochlear lateral wall. PLoS One. Jun. 20, 2017;12(6):e0179293. doi: 10.1371/journal.pone.0179293. eCollection 2017.
[cited by applicant]
Li et al., Notch inhibition induces mitotically generated hair cells in mammalian cochleae via activating the Wnt pathway. Proc Natl Acad Sci U S A. Jan. 6, 2015;112(1):166-71. doi: 10.1073/pnas.1415901112. Epub Dec. 22…
[cited by applicant]
Lin et al., Hearing loss and incident dementia. Arch Neurol. Feb. 2011;68(2):214-20. doi: 10.1001/archneurol.2010.362.
[cited by applicant]
Lin et al., Hearing loss prevalence in the United States. Arch Intern Med. Nov. 14, 2011;171(20):1851-2. doi: 10.1001/archinternmed.2011.506.
[cited by applicant]
Lukashkina et al., Amplification mode differs along the length of the mouse cochlea as revealed by connexin 26 deletion from specific gap junctions. Sci Rep. Jul. 12, 2017;7(1):5185. doi: 10.1038/s41598-017-04279-3.
[cited by applicant]
Lustig et al., Cochlear Gene Therapy. Cold Spring Harb Perspect Med. Sep. 3, 2019;9(9):a033191. doi: 10.1101/cshperspect.a033191.
[cited by applicant]
Ma et al., Connexin 26 in mature ears influences survival of hair cells and neurons. ARO Abstract 404. 2020; 43:258-9.
[cited by applicant]
Maguire et al., Efficacy, Safety, and Durability of Voretigene Neparvovec-rzyl in RPE65 Mutation-Associated Inherited Retinal Dystrophy: Results of Phase 1 and 3 Trials. Ophthalmology. Sep. 2019;126(9):1273-1285. doi: 1…
[cited by applicant]
Mahendrasingam et al., Subcellular distribution and relative expression of fibrocyte markers in the CD/1 mouse cochlea assessed by semiquantitative immunogold electron microscopy. J Histochem Cytochem. Nov. 2011;59(11):…
[cited by applicant]
Mason et al., Universal infant hearing screening by automated auditory brainstem response measurement. Pediatrics. Feb. 1998;101(2):221-8. doi: 10.1542/peds.101.2.221.
[cited by applicant]
McLean et al., GREAT improves functional interpretation of cis-regulatory regions. Nat Biotechnol. May 2010;28(5):495-501. doi: 10.1038/nbt.1630. Epub May 2, 2010.
[cited by applicant]
Mei et al., A deafness mechanism of digenic Cx26 (GJB2) and Cx30 (GJB6) mutations: Reduction of endocochlear potential by impairment of heterogeneous gap junctional function in the cochlear lateral wall. Neurobiol Dis. …
[cited by applicant]
Mutai et al., Mitotic activity and specification of fibrocyte subtypes in the developing rat cochlear lateral wall. Neuroscience. Nov. 10, 2009;163(4):1255-63. doi: 10.1016/j.neuroscience.2009.07.059. Epub Aug. 4, 2009.
[cited by applicant]
Narui et al., Tuning Inner-Ear Tip-Link Affinity Through Alternatively Spliced Variants of Protocadherin-15. Biochemistry. Mar. 20, 2018;57(11):1702-1710. doi: 10.1021/acs.biochem.7b01075. Epub Mar. 6, 2018.
[cited by applicant]
Nickel et al., Gap junctions and connexins in the inner ear: their roles in homeostasis and deafness. Curr Opin Otolaryngol Head Neck Surg. Oct. 2008;16(5):452-7. doi: 10.1097/MOO.0b013e32830e20b0.
[cited by applicant]
Olson et al., Post-translational tools expand the scope of synthetic biology. Curr Opin Chem Biol. Aug. 2012;16(3-4):300-6. doi: 10.1016/j.cbpa.2012.06.003. Epub Jul. 4, 2012.
[cited by applicant]
Paige et al., RNA mimics of green fluorescent protein. Science. Jul. 29, 2011;333(6042):642-6. doi: 10.1126/science.1207339.
[cited by applicant]
Pan et al., Gene therapy restores auditory and vestibular function in a mouse model of Usher syndrome type 1c. Nat Biotechnol. Mar. 2017;35(3):264-272. doi: 10.1038/nbt.3801. Epub Feb. 6, 2017.
[cited by applicant]
Pepermans et al., The CD2 isoform of protocadherin-15 is an essential component of the tip-link complex in mature auditory hair cells. EMBO Mol Med. Jul. 2014;6(7):984-92. doi: 10.15252/emmm.201403976.
[cited by applicant]
Plantier et al., A factor VIII minigene comprising the truncated intron I of factor IX highly improves the in vitro production of factor VIII. Thromb Haemost. Aug. 2001;86(2):596-603.
[cited by applicant]
Powers et al., A Partial Calcium-Free Linker Confers Flexibility to Inner-Ear Protocadherin-15. Structure. Mar. 7, 2017;25(3):482-495. doi: 10.1016/j.str.2017.01.014. Epub Feb. 23, 2017.
[cited by applicant]
Qi et al., Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Cell. Feb. 28, 2013;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.
[cited by applicant]
Rada-Iglesias et al., A unique chromatin signature uncovers early developmental enhancers in humans. Nature. Feb. 10, 2011;470(7333):279-83. doi: 10.1038/nature09692. Epub Dec. 15, 2010.
[cited by applicant]
Rees et al., Base editing: precision chemistry on the genome and transcriptome of living cells. Nat Rev Genet. Dec. 2018;19(12):770-788. doi: 10.1038/s41576-018-0059-1.
[cited by applicant]
Rodriguez-Paris et al., Comparative functional characterization of novel non-syndromic GJB2 gene variant p.Gly45Arg and lethal syndromic variant p.Gly45Glu. PeerJ. Oct. 11, 2016;4:e2494. doi: 10.7717/peerj.2494. eCollec…
[cited by applicant]
Sacheli et al., Gene transfer in inner ear cells: a challenging race. Gene Ther. Mar. 2013;20(3):237-47. doi: 10.1038/gt.2012.51. Epub Jun. 28, 2012.
[cited by applicant]
Scheffer et al., Gene expression profiling identifies Hes6 as a transcriptional target of ATOH1 in cochlear hair cells. FEBS Lett. Oct. 2, 2007;581(24):4651-6. doi: 10.1016/j.febslet.2007.08.059. Epub Sep. 4, 2007.
[cited by applicant]
Scheffer et al., The alphal subunit of nicotinic acetylcholine receptors in the inner ear: transcriptional regulation by ATOH1 and co-expression with the gamma subunit in hair cells. J Neurochem. Dec. 2007;103(6):2651-6…
[cited by applicant]
Scheffer et al., XIRP2, an actin-binding protein essential for inner ear hair-cell stereocilia. Cell Rep. Mar. 24, 2015;10(11):1811-8. doi: 10.1016/j.celrep.2015.02.042. Epub Mar. 12, 2015.
[cited by applicant]
Senis et al., CRISPR/Cas9-mediated genome engineering: an adeno-associated viral (AAV) vector toolbox. Biotechnol J. Nov. 2014;9(11):1402-12. doi: 10.1002/biot.201400046. Epub Oct. 6, 2014.
[cited by applicant]
Shepherd et al., The extent of adaptation in bullfrog saccular hair cells. J Neurosci. Oct. 1994;14(10):6217-29. doi: 10.1523/JNEUROSCI.14-10-06217.1994.
[cited by applicant]
Shu et al., Identification of Adeno-Associated Viral Vectors That Target Neonatal and Adult Mammalian Inner Ear Cell Subtypes. Hum Gene Ther. Sep. 2016;27(9):687-99. doi: 10.1089/hum.2016.053. Epub Jun. 24, 2016.
[cited by applicant]
Solc et al., Molecular cloning of myosins from the bullfrog saccular macula: A candidate for the adaptation motor. Auditory Neurosci. 1994; 1:63-75.
[cited by applicant]
Sotomayor et al., A Partial Calcium-Free Linker Confers Flexibility to Inner-Ear Protocadherin-15. Structure. Mar. 7, 2017;25(3):482-495. doi: 10.1016/j.str.2017.01.014. Epub Feb. 23, 2017.
[cited by applicant]
Sotomayor et al., In search of the hair-cell gating spring: Elastic properties of ankyrin and cadherin repeats. Structure. Apr. 2005;13(4):669-82. doi: 10.1016/j.str.2005.03.001.
[cited by applicant]
Sotomayor et al., Structural determinants of cadherin-23 function in hearing and deafness. Neuron. Apr. 15, 2010;66(1):85-100. doi: 10.1016/j.neuron.2010.03.028.
[cited by applicant]
Sun et al., Connexin30 null and conditional connexin26 null mice display distinct pattern and time course of cellular degeneration in the cochlea. J Comp Neurol. Oct. 20, 2009;516(6):569-79. doi: 10.1002/cne.22117.
[cited by applicant]
Takada et al., Connexin 26 null mice exhibit spiral ganglion degeneration that can be blocked by BDNF gene therapy. Hearing Res. Mar. 2014;309:124-35. doi: 10.1016/j.heares.2013.11.009. Epub Dec. 12, 2013.
[cited by applicant]
Trapani et al., Effective delivery of large genes to the retina by dual AAV vectors. EMBO Mol Med. Feb. 2014;6(2):194-211. doi: 10.1002/emmm.201302948. Epub Dec. 15, 2013.
[cited by applicant]
Trapani et al., Seeing the Light after 25 Years of Retinal Gene Therapy. Trends Mol Med. Aug. 2018;24(8):669-681. doi: 10.1016/j.molmed.2018.06.006. Epub Jul. 5, 2018.
[cited by applicant]
Trapani, Dual AAV Vectors for Stargardt Disease. Methods Mol Biol. 2018; 1715:153-175. doi: 10.1007/978-1-4939-7522-8_11.
[cited by applicant]
Truong et al., Development of an intein-mediated split-Cas9 system for gene therapy. Nucleic Acids Res. Jul. 27, 2015;43(13):6450-8. doi: 10.1093/nar/gkv601. Epub Jun. 16, 2015.
[cited by applicant]
Vogl, Tryptophan-rich basic protein (WRB) mediates insertion of the tail-anchored protein otoferlin and is required for hair cell exocytosis and hearing. EMBO J. Dec. 1, 2016;35(23):2536-2552. doi: 10.15252/embj.2015935…
[cited by applicant]
Wang et al., Targeted connexin26 ablation arrests postnatal development of the organ of Corti. Biochem Biophys Res Commun. Jul. 17, 2009;385(1):33-7. doi: 10.1016/j.bbrc.2009.05.023. Epub May 9, 2009.
[cited by applicant]
Wassmer et al., Exosome-associated AAV2 vector mediates robust gene delivery into the murine retina upon intravitreal injection. Sci Rep. Mar. 31, 2017;7:45329. doi: 10.1038/srep45329.
[cited by applicant]
Watanabe et al., Expression of the Sox10 gene during mouse inner ear development. Brain Res Mol Brain Res. Dec. 8, 2000;84(1-2):141-5. doi: 10.1016/s0169-328x(00)00236-9.
[cited by applicant]
Wilch et al., A novel DFNB1 deletion allele supports the existence of a distant cis-regulatory region that controls GJB2 and GJB6 expression. Clin Genet. Sep. 2010;78(3):267-74. doi: 10.1111/j.1399-0004.2010.01387.x. Ep…
[cited by applicant]
Wingard et al., Cellular and Deafness Mechanisms Underlying Connexin Mutation-Induced Hearing Loss—A Common Hereditary Deafness. Front Cell Neurosci. May 29, 2015;9:202. doi: 10.3389/fncel.2015.00202. eCollection 2015.
[cited by applicant]
Wise et al., The effect of deafness duration on neurotrophin gene therapy for spiral ganglion neuron protection. Hearing Res. Aug. 2011;278(1-2):69-76. doi: 10.1016/j.heares.2011.04.010. Epub May 1, 2011.
[cited by applicant]
Wu et al., Hair-cell mechanotransduction persists in TRP channel knockout mice. PloS One. May 19, 2016;11(5):e0155577. doi: 10.1371/journal.pone.0155577. eCollection 2016.
[cited by applicant]
Xia et al., Expression of connexin 26 and Na,K-ATPase in the developing mouse cochlear lateral wall: functional implications. Brain Res. Oct. 30, 1999;846(1):106-11. doi: 10.1016/s0006-8993(99)01996-4.
[cited by applicant]
Xiao et al., Rescue of the albino phenotype by introducing a functional tyrosinase minigene into Kunming albino mice. World J Gastroenterol. Jan. 14, 2007;13(2):244-9. doi: 10.3748/wjg.v13.i2.244.
[cited by applicant]
Yang et al., Gfi1-Cre knock-in mouse line: A tool for inner ear hair cell-specific gene deletion. Genesis. Jun. 2010;48(6):400-6. doi: 10.1002/dvg.20632.
[cited by applicant]
Yeh et al., In vivo base editing of post-mitotic sensory cells. Nature Commun. Jun. 5, 2018;9(1):2184. doi: 10.1038/s41467-018-04580-3.
[cited by applicant]
Yeh et al., In vivo base editing rescues hearing in a mouse model of recessive deafness. ARO Abstract 172. 2020; 43:96-97.
[cited by applicant]
Yu et al., Virally expressed connexin26 restores gap junction function in the cochlea of conditional Gjb2 knockout mice. Gene Ther. Jan. 2014;21(1):71-80. doi: 10.1038/gt.2013.59. Epub Nov. 14, 2013.
[cited by applicant]
Zelante et al., Connexin26 mutations associated with the most common form of non-syndromic neurosensory autosomal recessive deafness (DFNB1) in Mediterraneans. Hum Mol Genet. Sep. 1997;6(9):1605-9. doi: 10.1093/hmg/6.9.…
[cited by applicant]
Zhu et al., Active cochlear amplification is dependent on supporting cell gap junctions. Nature Commun. 2013;4:1786. doi: 10.1038/ncomms2806.
[cited by applicant]
Extended European Search Report for Application No. 20794633.6, mailed Sep. 27, 2023.
[cited by applicant]
International Preliminary Report on Patentability for Application No. PCT/US2021/050188, mailed Mar. 23, 2023.
[cited by applicant]
Ahmed et al., Gene structure and mutant alleles of PCDH15: nonsyndromic deafness DFNB23 and type 1 Usher syndrome. Hum Genet. Oct. 2008;124(3):215-23. doi: 10.1007/s00439-008-0543-3. Epub Aug. 22, 2008.
[cited by applicant]
Ivanchenko et al., Mini-PCDH15 gene therapy rescues hearing in a mouse model of Usher syndrome type 1F. Nat Commun. Apr. 26, 2023;14(1):2400. doi: 10.1038/s41467-023-38038-y.
[cited by applicant]
Pepermans et al., The tip-link molecular complex of the auditory mechano-electrical transduction machinery. Hear Res. Dec. 2015;330(Pt A):10-7. doi: 10.1016/j.heares.2015.05.005. Epub Jun. 3, 2015.
[cited by applicant]
Zhang et al., Cochlear Gene Therapy for Sensorineural Hearing Loss: Current Status and Major Remaining Hurdles for Translational Success. Front Mol Neurosci. Jun. 26, 2018:11:221. doi: 10.3389/fnmol.2018.00221. eCollect…
[cited by applicant]
Sotomayor et al., Structure of a force-conveying cadherin bond essential for inner-ear mechanotransduction. Nature. Dec. 6, 2012;492(7427):128-32. doi: 10.1038/nature11590. Epub Nov. 7, 2012.
[cited by applicant]
Genbank Submission. NCBI; Accession No. NG_009115, version NG_009115.1.
[cited by applicant]