Fig 2: One edited ARMS2 SNP rs10490924 accounts for the preferential efficacy of phenylbutyrate (NaPB) in iRPE survival. (A–C) ROS expression as measured by DHE fluorescence in different aging models. One-way ANOVA was performed to calculate significance. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. The risk SNP rs10490924 (T) in ARMS2 raises ROS levels in (A) aged 16-wk, (B) H2O2 (100 µM, 3 h), and (C) A2E and blue light–exposed iRPE-L to levels seen in iRPE-H. (D–F) The antioxidant NaPB (50 µM) attenuates ROS in ARMS2 rs10490924 iRPE-L1 than iRPE-L2, L3 in (D) aged 16-wk, (E) H2O2 (100 µM, 3 h), and (F) A2E and blue light–exposed AMD patient-derived iRPE cells. For D–F, multiple paired t test was performed to calculate significance. Each point represents the mean value from 3 samples of the same cell line, where black is cell line 1, blue is cell line 2, and red is cell line 3; error bars represent the SD.

Fig 3: One edited ARMS2 SNP rs10490924 accounts for the preferential efficacy of phenylbutyrate (NaPB) in iRPE survival. Analysis of cell viability in (A) aged 16-wk; (B) H2O2 (100 µM, 3 h) and (C) A2E plus blue light–exposed cells. One-way ANOVA was performed to calculate significance. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. (D–F) NaPB (50 µM) enhances cell viability in high risk (iRPE-H AMD) and SNV edited (iRPE-L, e1) under all treatments: aged 16 wk (D), H2O2-induced oxidative stress (100 µM for 3 h) (E), and oxidatively stressed A2E and blue light–associated oxidative stress (F). Cell viability was normalized to that of the iRPE low-risk cell lines without NaPB treatment. Multiple paired t test was performed to calculate significance. Each point represents the mean value from 3 samples of the same cell line where black is cell line 1, blue is cell line 2, and red is cell line 3; error bars represent the SD.

Fig 4: One edited ARMS2 SNP rs10490924 (T) raised ROS levels. (A) Progressive ROS accumulation due to the rs10490924 (T) risk allele in ARMS2. The risk SNP rs10490924 (T) in ARMS2 raises ROS levels in the iRPE. ROS in ARMS2 rs10490924-edited (iRPE-L, e1) and high-risk T/T iRPE-H were increased at weeks 6, 12, and 16. Mean values with SD are shown. Multiple unpaired t test was performed to determine significance of ROS as determined by the DHE fluorescence in each cell line. One edited ARMS2 SNP rs10490924 (T) reduced superoxide dismutase (SOD) activity in (B) aged 16-wk-old, (C) H2O2-treated (100 µM, 3 h), (D) A2E + blue light–treated iRPE-L to levels seen in iRPE-H. Editing of e1 disproportionately increases IL-6 (E) and IL-8 (F) mRNA levels relative to the editing of e2 and e3, bringing IL-6 and IL-8 from levels seen in low-risk cells to that seen in high-risk cells. Inflammatory IL-8 is an autocrine agent for microglia which induces chemotaxis of cells to sites of injury. For B–F, one-way ANOVA was performed to determine significance (*P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001). Each point represents the mean value from 3 samples of the same cell line where black is cell line 1, blue is cell line 2, and red is cell line 3; error bars represent the SD.