The Transcriptome Profile of Retinal Pigment Epithelium and Müller Cell Lines Protected by Risuteganib Against Hydrogen Peroxide Stress

Abstract

Purpose: Oxidative stress contributes to the pathogenesis of vision-impairing diseases. In the retina, retinal pigment epithelium (RPE) and Müller cells support neuronal homeostasis, but also contribute to pathological development under stressed conditions. Recent studies found that the investigational drug risuteganib (RSG) has a good safety profile, provided protection in experimental models, and improved visual acuity in patients. The present in vitro study evaluated the effects of RSG in RPE and Müller cell lines stressed with the oxidant hydrogen peroxide (H 2 O 2 ). Methods: Human RPE (ARPE-19) and Müller (MIO-M1) cell lines were treated with various combinations of RSG and H 2 O 2 . Trypan blue assay was used to investigate the effect of compounds on cell viability. Gene expression was measured using RNA sequencing to identify regulated genes and the biological processes and pathways involved. Results: Trypan blue assay found RSG pre-treatment significantly protected against H 2 O 2 -induced cell death in ARPE-19 and MIO-M1 cells. Transcriptome analysis found H 2 O 2 regulated genes in several disease-relevant biological processes, including cell adhesion, migration, death, and proliferation; ECM organization; angiogenesis; metabolism; and immune system processes. RSG pre-treatment modulated these gene expression profiles in the opposite direction of H 2 O 2 . Pathway analysis found genes in integrin, AP-1, and syndecan signaling pathways were regulated. Expression of selected RSG-regulated genes was validated using qRT-PCR. Conclusions: RSG protected cultured human RPE and Müller cell lines against H 2 O 2 -induced cell death and mitigated the associated transcriptome changes in biological processes and pathways relevant to the pathogenesis of retinal diseases. These results demonstrate RSG reduced oxidative stress-induced toxicity in two retinal cell lines with potential relevance to the treatment of human diseases.

Endogenous and Exogenous Regulation of Redox Homeostasis in Retinal Pigment Epithelium Cells: An Updated Antioxidant Perspective

“…Hyperglycemia is accompanied by an intracellular decrease in the concentration of H + ions, which results in acidification of the retina. Growth factor imbalance, increased production of pro-inflammatory cytokines and ROS [ 42 , 83 ], ATP secretion, and activation of ATP-dependent Ca 2+ channels (P2X7R) are signals of damage to RPE and retinal cells [ 31 , 84 ].…”

“…Genetic disorders that affect the functioning of transcription factors involved in the control of differentiation in embryogenesis lead to RPE dysfunction, the death of neurons in the outer nuclear layer and in photoreceptors (rods and cones), and the death of neurons in the inner nuclear layers of the retina [ 25 , 26 , 27 ]. Transcriptome analysis showed a high level of expression of regulatory redox-dependent genes that provide resistance to oxidative reactions in the RPE [ 28 , 29 , 30 , 31 ]. RPE homeostasis largely depends on the stability state of the Bruch’s membrane (BM) [ 1 ].…”