Optimization of AAV tools to target Muller glial cells for retinal gene therapy

Urrutia-Cabrera, D.; Huppert, G.; Chu, S.; Wang, L.; Nguy, C. B.; Liu, C. F.; Lisowki, L.; Luu, C.; Wang, J.-H.; Hung, S.; Hewitt, A. W.; Huang, C.-L.; Edwards, T.; Martin, K.; Wong, R.

doi:10.64898/2025.12.15.694492

bioRxiv · 10.64898/2025.12.15.694492

Optimization of AAV tools to target Muller glial cells for retinal gene therapy

Urrutia-Cabrera, D.·Huppert, G.·Chu, S.·Wang, L.·Nguy, C. B.·Liu, C. F.·Lisowki, L.·Luu, C.·Wang, J.-H.·Hung, S.·Hewitt, A. W.·Huang, C.-L.·Edwards, T.·Martin, K.·Wong, R.

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Abstract

Reprogramming of Muller glial (MG) cells into retinal neurons has the potential to treat vision loss by regenerating the retina. Development of efficient gene delivery systems to target the MG cells is critical. Adeno-associated virus (AAV) serotypes and promoter specificity are important factors that influence AAV transduction profile in the retina. However, studies that optimize these parameters to specifically target MG cells are limited, in particular in rats which are commonly used for eye research. Here we tested 4 AAV serotypes and 14 promoters to optimize gene delivery to human MG cells in vitro and/or rat MG cells in vivo. We showed that the combinatorial use of MG-specific serotypes and promoters achieved high specificity for MG cell targeting, with ShH10Y serotype and the GFAP (gfaABC1D) promoter as the best performing tool to target rat MG cells in vivo. We developed new AAV vectors using known and novel MG-specific promoters and engineered short promoter variants to improve the cargo capacity of AAV delivery. Our results highlighted a number of promoters that can target MG cells in vitro or in vivo. This study further expands the AAV toolbox to target MG cells, which has important implications for retinal gene therapy development.

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Urrutia-Cabrera, D., Huppert, G., Chu, S., Wang, L., Nguy, C. B., Liu, C. F., Lisowki, L., Luu, C., Wang, J.-H., Hung, S., Hewitt, A. W., Huang, C.-L., Edwards, T., Martin, K., Wong, R.. 2025-12-16. Optimization of AAV tools to target Muller glial cells for retinal gene therapy. https://doi.org/10.64898/2025.12.15.694492

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