BioRxiv. 2023 Aug 23
Longlong Zhang, Hui Wang, Mengzhao Xun, Honghai Tang, Jinghan Wang, Jun Lv, Biyun Zhu, Yuxin Chen, Daqi Wang, Shaowei Hu, Ziwen Gao, Jianping Liu, Zheng-Yi Chen, Bing Chen, Huawei Li and Yilai Shu
Products used in the paper Details Operation
AAV vector packaging The AAVs used in this study include AAV1-CAG-hOTOF NT-AK, AAV1-CAG-hOTOF NT-AP, AAV1-CAG-hOTOF
AAV1-hOTOF CT-TS and AAV1-Myo15-GFP-WPRE-bGH-polyA. The vector plasmid together with capsid and helper plasmids were transiently transfected into HEK293T cells to produce viral particles. The viruses were commissioned from
PackGene Biotech (Guangzhou, China).
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Research Field: hearing

AAV Serotype: AAV1

Dose: A total of 20 μL AAV1-GFP was microinjected into the scala tympani through the RWM.

Routes of Administration: inner ear injection

Targeted organ: ear

Animal or cell line strain: ROtof –/– mice


Pathogenic mutations in the OTOF gene cause autosomal recessive hearing loss 9 (DFNB9), one of the most common forms of auditory neuropathy. There is no biological treatment for DFNB9. Here, we designed an OTOF gene therapy agent by dual AAV1 carrying human OTOF coding sequences with the expression driven by the hair cell-specific promoter Myo15, AAV1-hOTOF. To develop a clinical application of AAV1-hOTOF gene therapy, we evaluated its efficacy and safety in animal models by pharmacodynamics, behavior, and histopathology. AAV1-hOTOF inner ear delivery significantly improved hearing in Otof −/− mice without affecting normal hearing in wild-type mice. AAV1 was predominately distributed to the cochlea although it was detected in other organs such as the central nervous system and the liver, and no obvious toxic effects of AAV1-hOTOF were observed in mice. To further evaluate the safety of Myo15 promoter-driven AAV1-transgene, AAV1-GFP was delivered into the inner ear of Macaca fascicularis via the round window membrane. AAV1-GFP transduced 60-94% of the inner hair cells along the cochlear turns. AAV1-GFP was detected in isolated organs and no significant adverse effects were detected. These results suggest that AAV1-hOTOF is well tolerated and effective in animals, providing critical support for its clinical translation.

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