Comparison of transfection effects of different serotypes of adeno-associated virus into inner hair cells of mouse cochlea by canalostomy

doi:10.3969/j.issn.1006-7795.2022.04.003

Journal of Capital Medical University ›› 2022, Vol. 43 ›› Issue (4): 527-534.doi: 10.3969/j.issn.1006-7795.2022.04.003

• Deafness Disease: Basic Research to Clinical Diagnosis and Treatment • Previous Articles Next Articles

Comparison of transfection effects of different serotypes of adeno-associated virus into inner hair cells of mouse cochlea by canalostomy

Liu Shan^1,2, Guo Rui^1,2, Song Xinyu^1,2, Yu Qianru^1,2, Chen Zhongrui^1,2, Liang Wenqi^1,2, Teng Qi^1,2, Gong Shusheng^1,2, Liu Ke^1,2*

1. Department of Otorhinolaryngology-Head and Neck Surgery, Beijing Friendship Hospital, Capital Medical University, Beijing 100050, China;
2. Clinical Center for Hearing Loss, Capital Medical University, Beijing 100050, China

Received:2022-04-29 Online:2022-08-21 Published:2022-10-28
Contact: *E-mail:liuke@ccmu.edu.cn
Supported by:
This study was supported by National Key Research and Development Program of China(2020YFC2005201),National Natural Science Foundation of China (82071037,81830030,81770997),Beijing Science and Technology Commission-Education Commission Joint Fund Project (KZ201810025040).

Abstract

Abstract: Objective To investigate the efficiency and safety of adeno-associated virus (AAV) of different serotypes transfected into inner ear cells of adult mice through posterior semicircular canal pathway. Methods Twenty C57BL/6J mice aged 6-8 weeks were randomly divided into five groups: AAV8 virus injection group, AAV9 virus injection group, AAVie virus injection group and Anc80L65 virus injection group. They were injected through the latter half regulatory path, and each mouse was injected with 2 μL of virus solution.In the normal control group, each mouse was injected with 2 μL normal saline, and the left ear was used as the operating ear. Auditory brainstem response (ABR) was examined one day before operation and two weeks after operation in each group, and then samples were taken for immunofluorescence staining to observe the distribution of virus in corti. Results AAV8 group: at 2 weeks after operation, 34.6%±1.8% inner hair cells were transfected at the apical and middle turn of cochlea, and 39%±5.9% inner hair cells at the middle and basal turn of cochlea expressed green fluorescent protein (GFP). AAV9 group: 92.5%±1.1% inner hair cells were transfected at the apical and middle turn of cochlea and 94.6%±1.0% inner hair cells expressed GFP at the middle and basal turn of cochlea 2 weeks after operation.AAVie group: 96.7%±1.5% inner hair cells were transfected at the apical and middle turn of the cochlea and 97.7%±0.8% inner hair cells at the middle and basal turn of the cochlea expressed GFP at 2 weeks after operation.Anc80L65 group: at 2 weeks after operation,62.5%±3.3% inner hair cells were transfected at the apical and middle turn of the cochlea, and 63.1%±6.1% inner hair cells at the middle and basal turn of the cochlea expressed GFP.No GFP expression was found in the normal control group. Conclusion Injecting AAVie and AAV9 virus solution through semicircular canal can efficiently transfect into cochlear inner hair cells. This result has certain reference significance for inner ear gene therapy of deafness.

Key words: adeno associated virus, virus transfection, cochlea, inner hair cells, posterior semicircular canal

CLC Number:

R764

Liu Shan, Guo Rui, Song Xinyu, Yu Qianru, Chen Zhongrui, Liang Wenqi, Teng Qi, Gong Shusheng, Liu Ke. Comparison of transfection effects of different serotypes of adeno-associated virus into inner hair cells of mouse cochlea by canalostomy[J]. Journal of Capital Medical University, 2022, 43(4): 527-534.

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Comparison of transfection effects of different serotypes of adeno-associated virus into inner hair cells of mouse cochlea by canalostomy

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