Hair Cell Transduction Efficiency of Single- and Dual-AAV Serotypes in Adult Murine Cochleae

• Published in: Molecular therapy. Methods & clinical development 2020-06, Vol.17, p.1167-1177
• Main Authors: Omichi, Ryotaro, Yoshimura, Hidekane, Shibata, Seiji B., Vandenberghe, Luk H., Smith, Richard J.H.
• Format: Article
• Language: English
• Subjects: AAV2; adeno-associated virus; deafness; dual vectors; gene therapy; hair cells; hearing loss; injection; tropism; viral vectors
• ISSN: 2329-0501; 2329-0501
• DOI: 10.1016/j.omtm.2020.05.007

Gene delivery is a key component for the treatment of genetic hearing loss. To date, a myriad of adeno-associated virus (AAV) serotypes and surgical approaches have been employed to deliver transgenes to cochlear hair cells, but the efficacy of dual transduction remains unclear. Herein, we investigated cellular tropism of single injections of AAV serotype 1 (AAV1), AAV2, AAV8, AAV9, and Anc80L65, and quantitated dual-vector co-transduction rates following co-injection of AAV2 and AAV9 vectors in adult murine cochlea. We used the combined round window membrane and canal fenestration (RWM+CF) injection technique for vector delivery. Single AAV2 injections were most robust and transduced 96.7% ± 1.1% of inner hair cells (IHCs) and 83.9% ± 2.0% of outer hair cells (OHCs) throughout the cochlea without causing hearing impairment or hair cell loss. Dual AAV2 injection co-transduced 96.9% ± 1.7% of IHCs and 65.6% ± 8.95% of OHCs. Together, RWM+CF-injected single or dual AAV2 provides the highest auditory hair cell transduction efficiency of the AAV serotypes we studied. These findings broaden the application of cochlear gene therapy targeting hair cells. [Display omitted] Transduction efficiency of inner and outer hair cells in the mature murine cochlea is excellent with adeno-associated virus serotype 2 (AAV2) when a round window membrane and canal fenestration technique is used. Dual-vector co-transduction of AAV2:AVV2 is equally robust and efficient, making AAV2 an excellent choice for single and/or dual applications that require hair cell transduction.