PCDH15 dual-AAV gene therapy for deafness and blindness in Usher syndrome type 1F models
Maryna V. Ivanchenko, … , Bence György, David P. Corey
Maryna V. Ivanchenko, … , Bence György, David P. Corey
Published October 23, 2024
Citation Information: J Clin Invest. 2024;134(23):e177700. https://doi.org/10.1172/JCI177700.
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Research Article Ophthalmology Otology

PCDH15 dual-AAV gene therapy for deafness and blindness in Usher syndrome type 1F models

Abstract

Usher syndrome type 1F (USH1F), resulting from mutations in the protocadherin-15 (PCDH15) gene, is characterized by congenital lack of hearing and balance, and progressive blindness in the form of retinitis pigmentosa. In this study, we explore an approach for USH1F gene therapy, exceeding the single AAV packaging limit by employing a dual–adeno-associated virus (dual-AAV) strategy to deliver the full-length PCDH15 coding sequence. We demonstrate the efficacy of this strategy in mouse USH1F models, effectively restoring hearing and balance in these mice. Importantly, our approach also proves successful in expressing PCDH15 protein in clinically relevant retinal models, including human retinal organoids and nonhuman primate retina, showing efficient targeting of photoreceptors and proper protein expression in the calyceal processes. This research represents a major step toward advancing gene therapy for USH1F and the multiple challenges of hearing, balance, and vision impairment.

Authors

Maryna V. Ivanchenko, Daniel M. Hathaway, Eric M. Mulhall, Kevin T.A. Booth, Mantian Wang, Cole W. Peters, Alex J. Klein, Xinlan Chen, Yaqiao Li, Bence György, David P. Corey

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Figure 4

Human PCDH15 exon structure and disease-associated variants.

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Human PCDH15 exon structure and disease-associated variants. (A) Gene sc...
(A) Gene schematic of PCDH15-CD2 and reported disease-causing variants. Three primary transcripts of PCDH15 (CD1, CD2, and CD3) translate 3 isoforms, which differ primarily in the 3′ end of the gene. Disease variants were gathered from the Deafness Variation Database, accessed August 2021. Disease variants are plotted based on their location on transcript NM_001142769 and color-coded based on associated disease phenotype: blue, non-syndromic hearing loss (DFNB23); red, Usher syndrome type 1F (USH1F); or green, non-syndromic retinitis pigmentosa (NSRP). Note that the variant p.P1796fs impacts the protein-coding sequence of only the CD1 isoform and has been reported to cause NSRP, suggesting that the CD1 splice form is required for retinal function but not hearing. Similarly, p.Q1576X in CD2 affects hearing but not vision. (B) Quantitative PCR evaluation of PCDH15-CD1, PCDH15-CD2, and PCDH15-CD3 expression in primate cochlea (n = 5) and retina (n = 6). One-way ANOVA followed by post hoc Tukey’s test was used to assess statistical significance. Data are presented as mean ± SEM. **P < 0.01, ***P < 0.001, ****P < 0.0001; n.s., not significant (P > 0.05).