Identifying a novel retinal pigment epithelium specific adeno-associated virus vector

Retinal Pigment Epithelium (RPE) are crucial for maintaining retinal homeostasis and the visual cycle. Currently, there are no effective treatments for blinding retinal diseases caused by RPE cell degeneration. Gene therapy presents a promising approach, but no gene therapy vectors specifically targeting RPE cells are available. In this study, we utilized an AAV2 random mutation library and high-throughput sequencing to screen AAV vectors (AAV206)capable of specifically targeting RPE cells. Intravitreal injection of the AAV206 vector in mice demonstrated that AAV206 specifically infected RPE cells, whereas AAV2 primarily infects the inner retina. Furthermore, AAV206 exhibited lower immunogenicity and toxicity compared to AAV2. Overexpression of sFLT-1 through AAV206 effectively inhibited the size and leakage of choroidal neovascularization. In summary, AAV206 is an ideal vector for gene therapy targeting RPE cells. Overall design: To assess retinal changes, mice aged 6-8 weeks were divided into two groups, with five mice per group. The mice received intravitreal injections of AAV2-GFP, and AAV206-GFP, respectively. Fourteen days after intravitreal injection, retinas were harvested for bulk RNA sequencing.