Engineering of the AAV-Compatible Hair Cell-Specific Small-Size Myo15 Promoter for Gene Therapy in the Inner Ear  

作　　者：Shao Wei Hu Jun Lv Zijing Wang Honghai Tang Hui Wang Fang Wang Daqi Wang Juan Zhang Longlong Zhang Qi Cao Yuxin Chen Ziwen Gao Yu Han Wuqing Wang Geng-lin Li Yilai Shu Huawei Li 

机构地区：[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 [2]Institute of Biomedical Science,Fudan University,Shanghai,200032,China [3]NHC Key Laboratory of Hearing Medicine(Fudan University),Shanghai,200032,China

出　　处：《Research》2024年第4期710-724,共15页研究(英文)

基　　金：supported by National Science Fund for Distinguished Young Scholars(82225014);the Science and Technology Commission of Shanghai Municipality(21JC1401000);the Ministry of Science and Technology of China(2021YFA1101302,2020YFA0908201);the National Natural Science Foundation of China(82192860,82171148,82201306);the China Postdoctoral Science Foundation(2022M720780);the Shanghai Municipal Health Commission(20224Z0003);the Science and Technology Commission of Shanghai Municipality(21S11905100 and 23J31900100);the Shanghai Municipal Education Commission(2023ZKZD12);the Shanghai Clinical Medical Research Center for Otolaryngology Diseases(20MC1920200);the science and technology innovation program of Hunan Province(2023RC4005);the Shuguang Program(20SG08)supported by Shanghai Municipal Education Commission and Shanghai Education Development Foundation,and Fudan University(yg2022-23).

摘　　要：Adeno-associated virus(AAV)-mediated gene therapy is widely applied to treat numerous hereditary diseases in animal models and humans.The specific expression of AAV-delivered transgenes driven by cell type-specific promoters should further increase the safety of gene therapy.However,current methods for screening cell type-specific promoters are labor-intensive and time-consuming.Herein,we designed a“multiple vectors in one AAV”strategy for promoter construction in vivo.Through this strategy,we truncated a native promoter for Myo15 expression in hair cells(HCs)in the inner ear,from 1,611 bp down to 1,157 bp,and further down to 956 bp.Under the control of these 2 promoters,green fluorescent protein packaged in AAV-PHP.eB was exclusively expressed in the HCs.The transcription initiation ability of the 2 promoters was further verified by intein-mediated otoferlin recombination in a dual-AAV therapeutic system.Driven by these 2 promoters,human otoferlin was selectively expressed in HCs,resulting in the restoration of hearing in treated Otof−/−mice for at least 52 weeks.In summary,we developed an efficient screening strategy for cell type-specific promoter engineering and created 2 truncated Myo15 promoters that not only restored hereditary deafness in animal models but also show great potential for treating human patients in future.

关 键 词：AAV MYO HEREDITARY 

分 类 号：R73[医药卫生—肿瘤]