Gene editing in a Myo6 semi-dominant mouse model rescues auditory function

Yuanyuan Xue; Xinde Hu; Daqi Wang; Di Li; Yige Li; Fang Wang; Mingqian Huang; Xi Gu; Zhijiao Xu; Jinan Zhou; Jinghan Wang; Renjie Chai; Jun Shen; Zheng-Yi Chen; Geng-Lin Li; Hui Yang; Huawei Li; Erwei Zuo; Yilai Shu

doi:10.1016/j.ymthe.2021.06.015

Gene editing in a Myo6 semi-dominant mouse model rescues auditory function

Mol Ther. 2022 Jan 5;30(1):105-118. doi: 10.1016/j.ymthe.2021.06.015. Epub 2021 Jun 24.

Authors

Yuanyuan Xue¹, Xinde Hu², Daqi Wang¹, Di Li³, Yige Li⁴, Fang Wang¹, Mingqian Huang⁵, Xi Gu¹, Zhijiao Xu¹, Jinan Zhou¹, Jinghan Wang¹, Renjie Chai⁶, Jun Shen⁷, Zheng-Yi Chen⁵, Geng-Lin Li⁸, Hui Yang⁹, Huawei Li¹⁰, Erwei Zuo¹¹, Yilai Shu¹²

Affiliations

¹ ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China; Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China; NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai 200031, China.
² Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Research Center for Brain Science and Brain-Inspired Intelligence, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
³ Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518124, China; State Key Lab for Conservation and Utilization of Subtropical Agric-Biological Resources, Guangxi University, Nanning 530005, China.
⁴ MOE Key Laboratory for Developmental Genes and Human Disease, School of Life Sciences and Technology, Jiangsu Province High-Tech Key Laboratory for Bio-Medical Research, Southeast University, Nanjing 210096, China.
⁵ Department of Otolaryngology-Head and Neck Surgery, Graduate Program in Speech and Hearing Bioscience and Technology and Program in Neuroscience, Harvard Medical School, Boston, MA 02115, USA; Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary, Boston, MA 02114, USA.
⁶ MOE Key Laboratory for Developmental Genes and Human Disease, School of Life Sciences and Technology, Jiangsu Province High-Tech Key Laboratory for Bio-Medical Research, Southeast University, Nanjing 210096, China; Co-Innovation Center of Neuroregeneration, Nantong University, Nantong 226001, China; Institute for Stem Cell and Regeneration, Chinese Academy of Science, Beijing, Shanghai 200032, China.
⁷ Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Harvard Medical School Center for Hereditary Deafness, Boston, MA 02115, USA.
⁸ ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China; NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai 200031, China.
⁹ College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
¹⁰ ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China; Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China; NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai 200031, China; The Institutes of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai 200032, China. Electronic address: hwli@shmu.edu.cn.
¹¹ Shenzhen Branch, Guangdong Laboratory for Lingnan Modern Agriculture, Genome Analysis Laboratory of the Ministry of Agriculture, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen 518124, China. Electronic address: erweizuo@163.com.
¹² ENT Institute and Department of Otorhinolaryngology, Eye & ENT Hospital, State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Fudan University, Shanghai 200031, China; Institutes of Biomedical Sciences, Fudan University, Shanghai 200032, China; NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai 200031, China. Electronic address: yilai_shu@fudan.edu.cn.

Abstract

Myosin VI（MYO6） is an unconventional myosin that is vital for auditory and vestibular function. Pathogenic variants in the human MYO6 gene cause autosomal-dominant or -recessive forms of hearing loss. Effective treatments for Myo6 mutation causing hearing loss are limited. We studied whether adeno-associated virus (AAV)-PHP.eB vector-mediated in vivo delivery of Staphylococcus aureus Cas9 (SaCas9-KKH)-single-guide RNA (sgRNA) complexes could ameliorate hearing loss in a Myo6^WT/C442Y mouse model that recapitulated the phenotypes of human patients. The in vivo editing efficiency of the AAV-SaCas9-KKH-Myo6-g2 system on Myo6^C442Y is 4.05% on average in Myo6^WT/C442Y mice, which was ∼17-fold greater than editing efficiency of Myo6^WT alleles. Rescue of auditory function was observed up to 5 months post AAV-SaCas9-KKH-Myo6-g2 injection in Myo6^WT/C442Y mice. Meanwhile, shorter latencies of auditory brainstem response (ABR) wave I, lower distortion product otoacoustic emission (DPOAE) thresholds, increased cell survival rates, more regular hair bundle morphology, and recovery of inward calcium levels were also observed in the AAV-SaCas9-KKH-Myo6-g2-treated ears compared to untreated ears. These findings provide further reference for in vivo genome editing as a therapeutic treatment for various semi-dominant forms of hearing loss and other semi-dominant diseases.

Keywords: CRISPR-Cas9; gene therapy; hearing loss; myosin VI; semi-dominant hearing loss.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Disease Models, Animal
Evoked Potentials, Auditory, Brain Stem / genetics
Gene Editing*
Hearing
Hearing Loss* / genetics
Hearing Loss* / therapy
Humans
Mice
RNA, Guide, CRISPR-Cas Systems

Substances

RNA, Guide, CRISPR-Cas Systems

Abstract

Publication types

MeSH terms

Substances

Grants and funding