PDF(Pubmed)
Abstract：
Adeno-associated viral (AAV) vectors are increasingly used as vehicles for gene delivery to treat hearing loss. However, lack of specificity of the transgene expression may lead to overexpression of the transgene in nontarget tissues. In this study, we evaluated the expression efficiency and specificity of transgene delivered by AAV-PHP.eB under the inner ear sensory cell-specific Myo15 promoter. Compared with the ubiquitous CAG promoter, the Myo15 promoter initiates efficient expression of the GFP fluorescence reporter in hair cells, while minimizing non-specific expression in other cell types of the inner ear and CNS. Furthermore, using the Myo15 promoter, we constructed an AAV-mediated therapeutic system with the coding sequence of OTOF gene. After inner ear injection, we observed apparent hearing recovery in Otof-/- mice, highly efficient expression of exogenous otoferlin, and significant improvement in the exocytosis function of inner hair cells. Overall, our results indicate that gene therapy mediated by the hair cell-specific Myo15 promoter has potential clinical application for the treatment of autosomal recessive deafness and yet for other hereditary hearing loss related to dysfunction of hair cells.
摘要：
腺相关病毒(AAV)载体越来越多地用作基因递送的载体以治疗听力损失。然而,转基因表达缺乏特异性可能导致转基因在非靶组织中过度表达。在这项研究中,我们评估了AAV-PHP传递的转基因的表达效率和特异性。内耳感觉细胞特异性Myo15启动子下的eB。与普遍存在的CAG启动子相比，Myo15启动子启动GFP荧光报告基因在毛细胞中的有效表达，同时将内耳和CNS的其他细胞类型中的非特异性表达降至最低。此外，使用Myo15启动子，我们构建了一个具有OTOF基因编码序列的AAV介导的治疗系统。内耳注射后,我们观察到Otof-/-小鼠明显的听力恢复，高效表达外源耳铁蛋白，内毛细胞的胞吐功能明显改善。总的来说,我们的结果表明,由毛细胞特异性Myo15启动子介导的基因治疗对于治疗常染色体隐性耳聋和其他与毛细胞功能障碍相关的遗传性耳聋具有潜在的临床应用价值.
