Figure 2.

rAAV-KH902 reduces laser damage-induced CNV in a prevention paradigm. (A) Time line of prevention paradigm as outlined in text. Laser damage (D) was performed on 4–8 different locations per retina (D0). Fundus and FFA were performed at D0 and over the next 15 days. (B) Representative FFA images of the same eye over time injected with vectors indicated. The number of leakage sites seen at D0 was used as denominator to calculate the percentage of remaining leakage sites over time. (C) Representative brightfield fundus images of the same eye over time injected with vectors indicated. Last column shows OCT analysis of CNV lesion at D15 (red arrowheads point to the same initial laser lesion). (D) Quantification of the percentage of the leakage sites remaining for each vector and time point indicated. The eGFP group shows combined data for all four eGFP vector serotypes. Number of initial leakage sites at D0: AAVs-eGFP (sum of all four eGFP serotypes) = 370, rAAV2-KH902 = 102, rAAV2.7m8-KH902 = 60, rAAV3b-KH902 = 119, and rAAV8-KH902 = 132. Bars show mean ± SEM (*p < 0.05, **p < 0.01, and ****p < 0.0001). (E) Representative RPE flat mounts stained for ZO-1 (ZO-1: RPE cell boundaries) and PECAM1 expression of eyes injected with indicated vectors, to identify neovascular lesions (red arrowheads). To the right, a higher magnification of PECAM1 staining (red) seen in the middle column, showing the remaining CNV sites. Scale bar = 500 μm (RPE flat mount) and 300 μm (CNV site) (ONH). (F) Example of output (green signal) generated by IMARIS software to quantify areas of CNV lesion. Scale bar = 300 μm. (G) Bar graph showing quantification of the average surface area of CNV lesion for serotypes indicated. AAVs-eGFP (average of all four eGFP serotypes). Values shown are mean ± SEM (**p < 0.01 and ****p < 0.0001). CNV, choroidal neovascularization; FFA, fundus fluorescein angiography; OCT, optical coherence tomography; ONH, optic nerve head; RPE, retinal-pigmented epithelial; ZO-1, zonula occludens 1.