Abstract
Mutations to the OTOF gene are among the most common reasons for auditory neuropathy. Although cochlear implants are often effective in restoring sound transduction, there are currently no biological treatments for individuals with variants of OTOF. Previous studies have reported the rescue of hearing in DFNB9 mice using OTOF gene replacement although the efficacy needs improvement. Here, we developed a novel dual-AAV-mediated gene therapy system based on the principles of protein trans-splicing, and we show that this system can reverse bilateral deafness in Otof –/– mice after a single unilateral injection. The system effectively expressed exogenous mouse or human otoferlin after injection on postnatal day 0–2. Human otoferlin restored hearing to near wild-type levels for at least 6 months and restored the release of synaptic vesicles in inner hair cells. Our study not only provides a preferential clinical strategy for the treatment of OTOF-related auditory neuropathies, but also describes a route of development for other large-gene therapies and protein engineering techniques.
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Data availability
The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
This work was supported by the National Key Research and Development Program of China (2020YFA0908201), the National Natural Science Foundation of China (82171148), the Clinical Research Plan of SHDC (SHDC2020CR4083), the Science and Technology Commission of Shanghai Municipality (21S11905100), the Special Project for Clinical Research in Health Industry of Shanghai Municipal Health Commission (20224Z0003), the Excellent Doctors-Excellent Clinical Researchers Program of Eye & ENT Hospital of Fudan University (SYB202005), and the “Shuguang Program” supported by the Shanghai Education Development Foundation and the Shanghai Municipal Education Commission (20SG08). We thank Shanghai Refreshgene Therapeutics Co., Ltd. for suggestions and assistance with the experiments.
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The National Key Research and Development Program of China, 2020YFA0908201, Yilai Shu, the National Natural Science Foundation of China, 82171148, Yilai Shu, the Clinical Research Plan of SHDC, SHDC2020CR4083, Yilai Shu, the Science and Technology Commission of Shanghai Municipality, 21S11905100, Yilai Shu, the Special Project for Clinical Research in Health Industry of Shanghai Municipal Health Commission, 20224Z0003, Yilai Shu, the Excellent Doctors-Excellent Clinical Researchers Program of Eye & ENT Hospital of Fudan University, SYB202005, Yilai Shu, the “Shuguang Program” supported by the Shanghai Education Development Foundation and the Shanghai Municipal Education Commission, 20SG08, Yilai Shu
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YS, HL, and WW: developed and supervised the project. HT and YS: jointly conceived the project. HT designed experiments and analyzed the data. HW, JL, MX, and FW: performed the in vivo experiments and the partial data analysis. HT: conducted the structural prediction. SW, HT, and SH: conducted the in vitro experiments. HT, YS, and SH: wrote the manuscript. HT, YS, SH, HW, SW, KG, YC, and DW: reviewed and revised the manuscript. All the authors read and approved the final manuscript.
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Tang, H., Wang, H., Wang, S. et al. Hearing of Otof-deficient mice restored by trans-splicing of N- and C-terminal otoferlin. Hum Genet 142, 289–304 (2023). https://doi.org/10.1007/s00439-022-02504-2
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DOI: https://doi.org/10.1007/s00439-022-02504-2