Xi Gu1,2,3,4, Daqi Wang1,2,4, Zhijiao Xu1,2,4, Yilai Shu5,6,7, Huawei Li8,9,10,11, Jinghan Wang1,2,4, Luo Guo1,2,4, Renjie Chai12,13,14, Genglin Li1,2,4. 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. Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China. 3. Department of Otolaryngology, the First Affiliated Hospital of Fujian Medical University, Fuzhou, 350005, China. 4. NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai, 200031, China. 5. 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. yilai_shu@fudan.edu.cn. 6. Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China. yilai_shu@fudan.edu.cn. 7. NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai, 200031, China. yilai_shu@fudan.edu.cn. 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. hwli@shmu.edu.cn. 9. Institutes of Biomedical Sciences, Fudan University, Shanghai, 200032, China. hwli@shmu.edu.cn. 10. NHC Key Laboratory of Hearing Medicine (Fudan University), Shanghai, 200031, China. hwli@shmu.edu.cn. 11. The Institutes of Brain Science and the Collaborative Innovation Center for Brain Science, Fudan University, Shanghai, 200032, China. hwli@shmu.edu.cn. 12. Key Laboratory for Developmental Genes and Human Disease, Ministry of Education, Institute of Life Sciences, Jiangsu Province High-Tech Key Laboratory for Bio-Medical Research, Southeast University, Nanjing, 210096, China. 13. Co-Innovation Center of Neuroregeneration, Nantong University, Nantong, 226001, China. 14. Institute for Stem Cell and Regeneration, Chinese Academy of Science, Beijing, China.
Abstract
BACKGROUND: Aging, noise, infection, and ototoxic drugs are the major causes of human acquired sensorineural hearing loss, but treatment options are limited. CRISPR/Cas9 technology has tremendous potential to become a new therapeutic modality for acquired non-inherited sensorineural hearing loss. Here, we develop CRISPR/Cas9 strategies to prevent aminoglycoside-induced deafness, a common type of acquired non-inherited sensorineural hearing loss, via disrupting the Htra2 gene in the inner ear which is involved in apoptosis but has not been investigated in cochlear hair cell protection. RESULTS: The results indicate that adeno-associated virus (AAV)-mediated delivery of CRISPR/SpCas9 system ameliorates neomycin-induced apoptosis, promotes hair cell survival, and significantly improves hearing function in neomycin-treated mice. The protective effect of the AAV-CRISPR/Cas9 system in vivo is sustained up to 8 weeks after neomycin exposure. For more efficient delivery of the whole CRISPR/Cas9 system, we also explore the AAV-CRISPR/SaCas9 system to prevent neomycin-induced deafness. The in vivo editing efficiency of the SaCas9 system is 1.73% on average. We observed significant improvement in auditory brainstem response thresholds in the injected ears compared with the non-injected ears. At 4 weeks after neomycin exposure, the protective effect of the AAV-CRISPR/SaCas9 system is still obvious, with the improvement in auditory brainstem response threshold up to 50 dB at 8 kHz. CONCLUSIONS: These findings demonstrate the safe and effective prevention of aminoglycoside-induced deafness via Htra2 gene editing and support further development of the CRISPR/Cas9 technology in the treatment of non-inherited hearing loss as well as other non-inherited diseases.
BACKGROUND: Aging, noise, infection, and ototoxic drugs are the major causes of human acquired sensorineural hearing loss, but treatment options are limited. CRISPR/Cas9 technology has tremendous potential to become a new therapeutic modality for acquired non-inherited sensorineural hearing loss. Here, we develop CRISPR/Cas9 strategies to prevent aminoglycoside-induced deafness, a common type of acquired non-inherited sensorineural hearing loss, via disrupting the Htra2 gene in the inner ear which is involved in apoptosis but has not been investigated in cochlear hair cell protection. RESULTS: The results indicate that adeno-associated virus (AAV)-mediated delivery of CRISPR/SpCas9 system ameliorates neomycin-induced apoptosis, promotes hair cell survival, and significantly improves hearing function in neomycin-treated mice. The protective effect of the AAV-CRISPR/Cas9 system in vivo is sustained up to 8 weeks after neomycin exposure. For more efficient delivery of the whole CRISPR/Cas9 system, we also explore the AAV-CRISPR/SaCas9 system to prevent neomycin-induced deafness. The in vivo editing efficiency of the SaCas9 system is 1.73% on average. We observed significant improvement in auditory brainstem response thresholds in the injected ears compared with the non-injected ears. At 4 weeks after neomycin exposure, the protective effect of the AAV-CRISPR/SaCas9 system is still obvious, with the improvement in auditory brainstem response threshold up to 50 dB at 8 kHz. CONCLUSIONS: These findings demonstrate the safe and effective prevention of aminoglycoside-induced deafness via Htra2 gene editing and support further development of the CRISPR/Cas9 technology in the treatment of non-inherited hearing loss as well as other non-inherited diseases.
Authors: Abdelghani El Rafei; Daniel C DeSimone; Aalap D Narichania; M Rizwan Sohail; Holenarasipur R Vikram; Zhuo Li; James M Steckelberg; Walter R Wilson; Larry M Baddour Journal: J Infect Date: 2018-06-30 Impact factor: 6.072
Authors: Han C Dan; Mei Sun; Satoshi Kaneko; Richard I Feldman; Santo V Nicosia; Hong-Gang Wang; Benjamin K Tsang; Jin Q Cheng Journal: J Biol Chem Date: 2003-11-25 Impact factor: 5.157
Authors: Lukas D Landegger; Bifeng Pan; Charles Askew; Sarah J Wassmer; Sarah D Gluck; Alice Galvin; Ruth Taylor; Andrew Forge; Konstantina M Stankovic; Jeffrey R Holt; Luk H Vandenberghe Journal: Nat Biotechnol Date: 2017-02-06 Impact factor: 54.908