RNA base editing therapy cures hearing loss induced by OTOF gene mutation

Yuanyuan Xue; Yong Tao; Xing Wang; Xueling Wang; Yilai Shu; Yuanhua Liu; Wen Kang; Sifan Chen; Zhenzhe Cheng; Boou Yan; Yanwei Xie; Lanting Bi; Haitao Jia; Jinhui Li; Qingquan Xiao; Liying Chen; Xuan Yao; Linyu Shi; Hui Yang; Hao Wu

doi:10.1016/j.ymthe.2023.10.019

RNA base editing therapy cures hearing loss induced by OTOF gene mutation

Mol Ther. 2023 Dec 6;31(12):3520-3530. doi: 10.1016/j.ymthe.2023.10.019. Epub 2023 Nov 2.

Authors

Yuanyuan Xue¹, Yong Tao², Xing Wang³, Xueling Wang², Yilai Shu⁴, Yuanhua Liu⁵, Wen Kang², Sifan Chen³, Zhenzhe Cheng², Boou Yan³, Yanwei Xie³, Lanting Bi³, Haitao Jia³, Jinhui Li³, Qingquan Xiao³, Liying Chen³, Xuan Yao³, Linyu Shi³, Hui Yang⁶, Hao Wu⁷

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China; HuidaGene Therapeutics Co., Ltd., Shanghai 200131, China.
² Department of Otolaryngology-Head and Neck Surgery, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China; Ear Institute, Shanghai Jiaotong University School of Medicine, Shanghai 200125, China; Shanghai Key Laboratory of Translational Medicine on Ear and Nose Diseases, Shanghai 200125, China.
³ HuidaGene Therapeutics Co., Ltd., Shanghai 200131, China.
⁴ ENT Institute and Department of Otorhinolaryngology, Eye and ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China; Institute of Biomedical Science, Fudan University, Shanghai 200032, China; NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai 200032, China.
⁵ Institute of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China.
⁶ HuidaGene Therapeutics Co., Ltd., Shanghai 200131, China; Institute of Neuroscience, Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai 200031, China. Electronic address: huiyang@huidagene.com.
⁷ Department of Otolaryngology-Head and Neck Surgery, Shanghai Ninth People's Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200011, China; Ear Institute, Shanghai Jiaotong University School of Medicine, Shanghai 200125, China; Shanghai Key Laboratory of Translational Medicine on Ear and Nose Diseases, Shanghai 200125, China. Electronic address: wuhao@shsmu.edu.cn.

PMID: 37915172
PMCID: PMC10727966 (available on 2024-12-06)
DOI: 10.1016/j.ymthe.2023.10.019

Abstract

Otoferlin (OTOF) gene mutations represent the primary cause of hearing impairment and deafness in auditory neuropathy. The c.2485C>T (p. Q829X) mutation variant is responsible for approximately 3% of recessive prelingual deafness cases within the Spanish population. Previous studies have used two recombinant AAV vectors to overexpress OTOF, albeit with limited efficacy. In this study, we introduce an enhanced mini-dCas13X RNA base editor (emxABE) delivered via an AAV9 variant, achieving nearly 100% transfection efficiency in inner hair cells. This approach is aimed at treating OTOF^Q829X, resulting in an approximately 80% adenosine-to-inosine conversion efficiency in humanized Otof^Q829X/Q829X mice. Following a single scala media injection of emxABE targeting OTOF^Q829X (emxABE-T) administered during the postnatal day 0-3 period in Otof^Q829X/Q829X mice, we observed OTOF expression restoration in nearly 100% of inner hair cells. Moreover, auditory function was significantly improved, reaching similar levels as in wild-type mice. This enhancement persisted for at least 7 months. We also investigated P5-P7 and P30 Otof^Q829X/Q829X mice, achieving auditory function restoration through round window injection of emxABE-T. These findings not only highlight an effective therapeutic strategy for potentially addressing OTOF^Q829X-induced hearing loss but also underscore emxABE as a versatile toolkit for treating other monogenic diseases characterized by premature termination codons.

Keywords: OTOF; RNA base editing; emxABE; gene therapy; hearing loss; otoferlin.

MeSH terms

Animals
Deafness*
Gene Editing
Hearing Loss* / genetics
Hearing Loss* / therapy
Hearing Loss, Central*
Mice
Mutation