Approaches and Vectors for Efficient Cochlear Gene Transfer in Adult Mouse Models

Yu Zhao; Longlong Zhang; Daqi Wang; Bing Chen; Yilai Shu

doi:10.3390/biom13010038

Approaches and Vectors for Efficient Cochlear Gene Transfer in Adult Mouse Models

Biomolecules. 2022 Dec 26;13(1):38. doi: 10.3390/biom13010038.

Authors

Yu Zhao^{1

2

3}, Longlong Zhang^{1

2

3}, Daqi Wang^{1

2

3}, Bing Chen^{1

2

3}, Yilai Shu^{1

2

3}

Affiliations

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 NHC Key Laboratory of Hearing Medicine, Fudan University, Shanghai 200031, China.

Abstract

Inner ear gene therapy using adeno-associated viral vectors (AAVs) in neonatal mice can alleviate hearing loss in mouse models of deafness. However, efficient and safe transgene delivery to the adult mouse cochlea is critical for the effectiveness of AAV-mediated therapy. Here, we examined three gene delivery approaches including posterior semicircular canal (PSCC) canalostomy, round window membrane (RWM) injection, and tubing-RWM+PSCC (t-RP) in adult mice. Transduction rates and survival rates of cochlear hair cells were analyzed, hearing function was recorded, AAV distribution in the sagittal brain sections was evaluated, and cochlear histopathologic images were appraised. We found that an injection volume of 1 μL AAV through the PSCC is safe and highly efficient and does not impair hearing function in adult mice, but local injection allows AAV vectors to spread slightly into the brain. We then tested five AAV serotypes (PHP.eB, IE, Anc80L65, AAV2, and PHP.s) in parallel and observed the most robust eGFP expression in inner hair cells, outer hair cells, and spiral ganglion neurons throughout the cochlea after AAV-Anc80L65 injection. Thus, PSCC-injected Anc80L65 provides a foundation for gene therapy in the adult cochlea and will facilitate the development of inner ear gene therapy.

Keywords: AAV; PSCC; RWM; adult mouse; gene therapy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cochlea / metabolism
Gene Transfer Techniques
Genetic Therapy / methods
Genetic Vectors* / genetics
Hearing Loss* / therapy
Mice

Grants and funding

This study was funded by grants from the National Key R&D Program of China, grant number 2020YFA0908201, the National Natural Science Foundation of China, grant number 82171148, 82225014, 82071048, 81870726, the Special Project for Clinical Research in Health Industry of Shanghai Municipal Health Commission, grant number 20224Z0003, the Science and Technology Commission of Shanghai Municipality, grant number 21S11905100, the Excellent physician—Excellent clinical researcher Plan of Eye & ENT Hospital of Fudan University, grant number SYB202005, the “Shuguang Program” supported by the Shanghai Education Development Foundation and the Shanghai Municipal Education Commission, grant number 20SG08.