Safety and efficacy of AAV8-aflibercept in treating choroidal neovascularization via single-cell RNA sequencing.

This study evaluated the efficacy and safety of adeno-associated virus serotype 8 (AAV8)-aflibercept for treating choroidal neovascularization (CNV) in mice model, utilizing single-cell RNA sequencing (scRNA-seq) to investigate retinal effects. AAV8-aflibercept was administered via subretinal injection to mice, followed by laser-induced CNV modeling. Successful expression was achieved following subretinal injection of AAV8-aflibercept, aflibercept protein expression showed a gradual decline after peaking at week 4, but remained at relatively stable levels through weeks 8-24. The therapeutic effect was assessed using fluorescein angiography and quantified. AAV8-aflibercept significantly decreased CNV areas across dosages from 3 × 10(7) to 1 × 10(9) GC/eye, with minor retinal thinning and retinal pigment epithelium (RPE) alterations observed only at dosages exceeding 1 × 10(9) GC/eye. Twelve distinct cell types were identified by single-cell RNA sequencing (scRNA-seq). RPE cells were the primary target, with 29.7% expressing the transduced gene. Target gene expression in RPE cells mainly involved RNA splicing and mRNA processing, with no strong activation signatures were observed in inflammation, apoptosis, or autophagy pathways. Upregulation of Gadd45b, Lrrc2, Lcn2 was noted in the AAV8-aflibercept group, suggesting a stress response. AAV8-aflibercept demonstrated significant efficacy and safety in treating CNV, with scRNA-seq providing insights into cellular changes.