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 2

Dual-AAV delivery of PCDH15 successfully preserves hearing in Myo15a-Cre mouse model and alleviates vestibular deficits in constitutive knockout model of USH1F.

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Dual-AAV delivery of PCDH15 successfully preserves hearing in Myo15a-Cre...
(A) Dual AAV vectors encoding PCDH15-CD2 protein were injected via the RWM into either conditional or constitutive knockout mice at P1. At 5 weeks, treated animals were assayed for hearing and vestibular function and processed for histology. (B) ABR click and tone thresholds for untreated Pcdh15fl/fl Cre control mice (n = 9), untreated Pcdh15fl/fl, Myo15a-Cre+/− mice (n = 10), and treated Pcdh15fl/fl, Myo15a-Cre+/− mice (n = 23). (C) Average DPOAE thresholds and amplitudes at 16 kHz for 5-week-old hearing control mice (n = 9), for untreated Pcdh15fl/fl, Myo15a-Cre+/− mice (n = 6), and treated Pcdh15fl/fl, Myo15a-Cre+/− mice (n = 22). (D) Representative open field path outlines of wild-type mice, untreated Pcdh15fl/fl, Myo15a-Cre+/− mice, Pcdh15–/– knockout mice, and treated Pcdh15–/– knockouts. (E) Summary of total rotations in wild type (n = 5), untreated Pcdh15fl/fl, Myo15a-Cre+/− mice (n = 5), Pcdh15–/– mice (n = 4), and treated Pcdh15–/– mice (n = 13). Untreated Pcdh15–/– mice exhibited severe circling behavior, while vestibular function was fully preserved at normal levels in dual AAV-treated Pcdh15–/– mice. (F) Wild-type (n = 5) and Pcdh15fl/fl, Myo15a-Cre+/− (n = 5) mice swam well. Pcdh15–/– mice (n = 4) were unable to swim normally. G) Pcdh15–/– mice showed much shorter latency to fall on the rotarod test (n = 4), while treated Pcdh15–/– mice performed at wild-type levels (n = 12). One-way ANOVA followed by Dunnett’s test was used to assess statistical significance. Data are presented as mean ± SEM. ***P < 0.001, ****P < 0.0001; n.s., not significant (P > 0.05).