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.
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.
Mol Ther. 2022 Jan 5;30(1):105-118. doi: 10.1016/j.ymthe.2021.06.015. Epub 2021 Jun 24.
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 mouse model that recapitulated the phenotypes of human patients. The in vivo editing efficiency of the AAV-SaCas9-KKH-Myo6-g2 system on Myo6 is 4.05% on average in Myo6 mice, which was ∼17-fold greater than editing efficiency of Myo6 alleles. Rescue of auditory function was observed up to 5 months post AAV-SaCas9-KKH-Myo6-g2 injection in Myo6 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.
肌球蛋白 VI（MYO6）是一种非传统的肌球蛋白，对听觉和前庭功能至关重要。人类 MYO6 基因的致病性变异导致常染色体显性或隐性形式的听力损失。针对 Myo6 突变引起的听力损失的有效治疗方法有限。我们研究了腺相关病毒（AAV）-PHP.eB 载体介导的金黄色葡萄球菌 Cas9（SaCas9-KKH）-单指导 RNA（sgRNA）复合物在体内递送是否可以改善 Myo6 小鼠模型中的听力损失，该模型模拟了人类患者的表型。AAV-SaCas9-KKH-Myo6-g2 系统在 Myo6 小鼠中的体内编辑效率平均为 4.05%，比 Myo6 等位基因的编辑效率高约 17 倍。在 AAV-SaCas9-KKH-Myo6-g2 注射后 5 个月观察到听觉功能的恢复。同时，与未治疗的耳朵相比，在 AAV-SaCas9-KKH-Myo6-g2 处理的耳朵中也观察到听觉脑干反应（ABR）波 I 的潜伏期更短、畸变产物耳声发射（DPOAE）阈值更低、细胞存活率更高、毛束形态更规则以及钙内流的恢复。这些发现为体内基因组编辑作为各种半显性形式的听力损失和其他半显性疾病的治疗方法提供了进一步的参考。
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