Mini-Pcdh15b Gene Therapy Rescues Visual Deficits in a Zebrafish Model of Usher Syndrome Type 1F

Chen, X.; Hathaway, D. M.; Klein, A. J.; Phillips, J. B.; Booth, K. T.; Heitsch, M.; Lytvyn, A.; Derfler, B. H.; Wegner, J.; Goldstein, C.; Murphy, A.; Megason, S. G.; Westerfield, M.; Corey, D. P.; Ivanchenko, M. V.

doi:10.1101/2025.11.05.686814

bioRxiv · 10.1101/2025.11.05.686814

Mini-Pcdh15b Gene Therapy Rescues Visual Deficits in a Zebrafish Model of Usher Syndrome Type 1F

Chen, X.·Hathaway, D. M.·Klein, A. J.·Phillips, J. B.·Booth, K. T.·Heitsch, M.·Lytvyn, A.·Derfler, B. H.·Wegner, J.·Goldstein, C.·Murphy, A.·Megason, S. G.·Westerfield, M.·Corey, D. P.·Ivanchenko, M. V.

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Abstract

Usher syndrome type 1F (USH1F) is a severe inherited disorder caused by mutations in PCDH15, resulting in congenital deafness, vestibular dysfunction, and progressive retinal degeneration leading to blindness. While cochlear implantation can restore hearing, no therapeutic interventions currently exist for vision loss. Gene augmentation therapy represents a promising approach; however, the PCDH15 coding sequence ([~]5.3 kb) exceeds the packaging capacity of adeno-associated virus (AAV) vectors. To overcome this limitation, we previously engineered shortened "mini-PCDH15" constructs that retain key structural and functional domains while fitting within a single AAV. Among these, the mini-PCDH15-V4 variant successfully restored hearing in Pcdh15-deficient mice. Here, we investigated the ability of a cone-targeted mini-pcdh15b-V4 transgene to rescue vision in a zebrafish model of USH1F. Untreated pcdh15b-deficient zebrafish exhibited severe structural and functional defects of the retina, including disorganized and shortened photoreceptor outer segments, disrupted calyceal processes, and markedly reduced electroretinogram (ERG) and optokinetic response (OKR) performance. Targeted expression of mini-pcdh15b-V4 recapitulated typical localization of Pcdh15b to calyceal processes and outer segment membranes, rescued photoreceptor architecture, and re-established both structural organization and functional output. Treated mutants exhibited improved visual tracking behavior and full recovery of ERG a-wave and b-wave amplitudes, indicating restoration of photoreceptor and synaptic function. Importantly, mini-pcdh15b-V4 expression produced no adverse effects in wild-type or heterozygous fish, supporting the safety of cone-specific expression. Together, these findings demonstrate that mini-pcdh15b-V4 can restore both photoreceptor structure and visual function in pcdh15b-deficient zebrafish. This work establishes the pcdh15b mutant zebrafish as a powerful preclinical model for studying USH1F retinopathy and supports the translational potential of rationally engineered mini-PCDH15 constructs as a feasible gene therapy approach for preventing or reversing vision loss in individuals with USH1F.

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Chen, X., Hathaway, D. M., Klein, A. J., Phillips, J. B., Booth, K. T., Heitsch, M., Lytvyn, A., Derfler, B. H., Wegner, J., Goldstein, C., Murphy, A., Megason, S. G., Westerfield, M., Corey, D. P., Ivanchenko, M. V.. 2025-11-06. Mini-Pcdh15b Gene Therapy Rescues Visual Deficits in a Zebrafish Model of Usher Syndrome Type 1F. https://doi.org/10.1101/2025.11.05.686814

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