Advances in the discovery of the causes of monogenic retinal disorders, combined with technologies for the delivery of DNA to the retina, offer enormous opportunities for the treatment of previously untreatable blinding diseases. However, for gene augmentation to be most effective, vectors that have the correct cell-type specificity are needed. While animal models are very useful, they often exhibit differences in retinal cell surface receptors compared to human retina. In this study, we evaluated the use of an ex vivo organotypic explant system to test the transduction efficiency and tropism of 7 different adeno-associated viral type 2 (AAV2) serotypes in human retina and retinal pigment epithelium-choroid: AAV2/1, AAV2/2, AAV2/4, AAV2/5, AAV2/6, AAV2/8, and AAV2/9, all driving expression of GFP under control of the cytomegalovirus promoter. After 7 days in culture, we found that AAV2/4 and AAV2/5 are particularly efficient at transducing photoreceptor cells and that AAV2/5 is highly specific to the outer nuclear layer. To validate the authenticity of the organotypic culture system, we also compared the transduction of the same set of AAVs in a pig model, in which subretinal injections in vivo were compared to cultured and transduced organotypic cultures ex vivo. This study shows how different AAV serotypes behave in the human retina and will provide insight for further investigation of each of these serotypes for gene augmentation-based treatment of inherited retinal degeneration.
Assessment of AAV Serotype Tropism in Human Retinal Explants.
Human Gene Therapy