Biomolecules. 2022 Dec 26
Yu Zhao, Longlong Zhang, Daqi Wang, Bing Chen, and Yilai Shu
Products used in the paper Details Operation
AAV vector packaging AAV-PHP.eB, AAV-ie, AAV-Anc80L65, AAV2, and AAV-PHP.s were produced by the PackGene Biotech Company (Shanghai, China). All vectors were engineered to express the enhanced green fluorescent protein (eGFP) gene under the control of the chimeric cytomegalovirus (CMV) promoter and carried the woodchuck hepatitis post-transcriptional virus regulatory element (WPRE) cassette. To determine the transduction rate among the different serotypes, we adjusted all the vectors to be at equal doses of 1.0×10^13 VG/mL. Virus aliquots were stored at -80 C and thawed before use. Request Quote

Research Field: ear

Keywords: gene therapy, AAV, adult mouse, PSCC, RWM

AAV Serotype: AAV-PHP.eB, AAV-ie, AAV-Anc80L65, AAV2, and AAV-PHP.s

Dose: 1×10^13 vg/ml

Routes of Administration: Canalostomy was first used to deliver AAVs into the mature cochlea through the PSCC approach. For the two experimental groups, a total of 1ul or 2ul AAV vectors were injected into the cochlea through the polymide tube at a rate of 5 nL/s.

Targeted organ: inner ear

Animal or cell line strain: Adult C57BL/6J wild-type mice (4–6 weeks old)


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.

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