Molecular Otolaryngology and Renal Research Laboratories, Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA.
Department of Otorhinolaryngology, Shinshu University School of Medicine, Matsumoto, Nagano, 390-8621, Japan.
Sci Rep. 2018 Feb 14;8(1):2980. doi: 10.1038/s41598-018-21233-z.
Cochlear gene therapy holds promise for the treatment of genetic deafness. Assessing its impact in adult murine models of hearing loss, however, has been hampered by technical challenges that have made it difficult to establish a robust method to deliver transgenes to the mature murine inner ear. Here in we demonstrate the feasibility of a combined round window membrane injection and semi-circular canal fenestration technique in the adult cochlea. Injection of both AAV2/9 and AAV2/Anc80L65 via this approach in P15-16 and P56-60 mice permits robust eGFP transduction of virtually all inner hair cells throughout the cochlea with variable transduction of vestibular hair cells. Auditory thresholds are not compromised. Transduction rate and cell tropism is primarily influenced by viral titer and AAV serotype but not age at injection. This approach is safe, versatile and efficient. Its use will facilitate studies using cochlear gene therapy in murine models of hearing loss over a wide range of time points.
耳蜗基因治疗有望治疗遗传性耳聋。然而，在成年小鼠听力损失模型中评估其影响受到技术挑战的阻碍，这些技术挑战使得难以建立一种将转基因递送到成熟的小鼠内耳的稳健方法。在这里，我们证明了在成年耳蜗中通过圆窗膜注射和半规管开窗联合技术的可行性。通过这种方法在 P15-16 和 P56-60 小鼠中注射 AAV2/9 和 AAV2/Anc80L65，允许实质上所有内毛细胞的 GFP 转导，在内耳中具有可变的前庭毛细胞转导。听觉阈值不受影响。转导率和细胞嗜性主要受病毒滴度和 AAV 血清型的影响，但不受注射时的年龄影响。这种方法安全、多功能且高效。它的使用将促进在广泛的时间点范围内使用耳蜗基因治疗在听力损失的小鼠模型中的研究。
