Figure 3.

AAV5-hGRK1-GUCY2D produced via PCL or TTx significantly improves retinal function in subretinally injected GCDKO mice

(A) Rod-mediated (scotopic) and cone-mediated (photopic) function were evaluated in GCDKO mice for 3 months after injection with producer cell line (PCL)- or triple transfection (TTx)-made vectors at a matched concentration of 1.5 × 10¹² vg/mL. This corresponds to a total dose of 1.5 × 10⁹ vg/eye. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, PCL versus uninjected Control; ˆp < 0.05, ˆˆp < 0.01, ˆˆˆp < 0.001, ˆˆˆˆp < 0.0001, PCL versus TTx; #p < 0.05, ##p < 0.01, ###p < 0.001, ####p < 0.0001, TTx versus uninjected control, as determined by two-way ANOVA with Sidak’s post-test analysis. Note than animal numbers measured over the course of the study are presented as a range in the legend. (B) Boundary sedimentation velocity profiles of AAV5 vectors containing the GUCY2D vector genome. Analytical ultracentrifugation (AUC) revealed that the PCL-made vector contained 81% full and 5% empty capsids, whereas the TTx-made vector contained 33% full and 45% empty capsids. (C) Mean ONL thickness in GCDKO mouse retinas treated with low- or mid-dose vectors (PCL or TTx made) were not significantly reduced relative to untreated controls. Injection with high-concentration (1 × 10¹³ vg/mL) PCL-made vector (corresponding to 1 × 10¹⁰ vg/eye) resulted in significant retinal thinning in treated animals. ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001, as determined by two-way ANOVA with Tukey’s post-test.