Effects of Vabysmo (Faricimab) on TBH-induced oxidative stressed ARPE-19 cells.

Abstract

Purpose : Age-related Macular Degeneration (AMD), a chronic, debilitating, retinal disease, is a leading cause of irreversible blindness worldwide. In recent decades, anti-vascular endothelial growth factor therapy (anti-VEGF) has been the standard option for neovascular AMD. Faricimab under the brand name of Vabysmo is a specific monoclonal antibody with dual effect of anti-VEGF-A and anti-Angiopoietin 2. The aim of this study is to evaluate the effects of Vabysmo on oxidatively stressed ARPE19 cells by measuring levels of cellular metabolism and reactive oxygen species (ROS).

Methods : The ARPE-19 cells were purchased from ATCC and cultured in DMEM-F12 media containing 50 µg/mL gentamycin, 17.5 mM glucose, 100 µg/mL streptomycin, 10% dialyzed fetal bovine serum, 100 unit/mL penicillin, 2.5 µg/mL fungizone, and 50 µg/mL gentamycin. Oxidative stress was induced by Tert-butyl hydroperoxide (tBH 125 μmol). Cells were plated in 96-well plates (10^4/well), cultured 24h and then treated 24h with DMSO (vehicle control), tBH alone or Vabysmo (Genentech, CA, USA) alone. The fourth group was pretreated with Vabysmo (24hr), then treated with tBH plus Vabysmo [tBH+Vaby] for another 48h. Cells were analyzed for cytotoxicity (Cytotox, IncuCyte,); cellular metabolism (MTT assay) and ROS (H2DCFDA assay). Data were analyzed Statistically via One Way-ANOVA and P values < 0.05 were considered significant.

Results :
The TBH-treated cells showed decreased cellular metabolism 57% (p-value <0.0001) compared with vehicle control. The [tBH+Vaby] cell had increased metabolism compared to tBH-treated cells (72%, p-value <0.05). The ROS levels were significantly decreased in [tBH+Vaby] cells (67%) compared with tBH alone (94%, p<0.0001). The InCucyte derived data confirmed the MTT and ROS results. Viable cell numbers were increased significantly after 48h treatment [tBH+Vaby]-treated cells (61%) when compared with the cells treated with tBH alone (47%, p-value <0.05).

Conclusions :
Vabysmo as an anti-VEGF-Angiopoietin 2 drug, increased cell viability and suppressed ROS level in tBH-treated ARPE-19 cells. Consequently, Vabysmo may protect RPE cells from oxidative stress caused by retinal diseases such as AMD.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.