Effects of Antiangiogenic Drugs on Expression Patterns of Epigenetic Pathway Genes.

BACKGROUND AND OBJECTIVE: To investigate the effects of antiangiogenic drugs on the transcription profile of acetylation genes in immortalized human retinal pigment epithelium cells (ARPE-19) in vitro. MATERIALS AND METHODS: This in vitro study evaluated the effect of antiangiogenic drugs on the expression of histone acetylation genes on immortalized ARPE-19 cell cultures. ARPE-19 cells were cultured, plated, and treated for 24 hours with aflibercept (Eylea; Regeneron, Tarrytown, NY), ranibizumab (Lucentis; Genentech, South San Francisco, CA), or bevacizumab (Avastin; Genentech, South San Francisco, CA) at one (1×) or two times (2×) the concentrations of the clinical intravitreal dose. Untreated cells were used as controls. RNA was isolated, and real-time quantitative reverse transcription polymerase chain reaction analysis was performed on individual samples to quantify expression levels of genes associated with epigenetic acetylation pathways: histone acetyltransferase 1 (HAT1) and histone deacetylases 1, 6, and 11 (HDAC1, HDAC6, and HDAC11). Differences in cycle thresholds (ΔΔCts) were obtained, and folds were calculated using the formula 2^ΔΔCt. Main outcome measures were expression levels of candidate genes in treated versus untreated samples. RESULTS: Compared with untreated cells, 1× ranibizumab-treated cells expressed higher levels of HDAC6, and 2× ranibizumab-treated cells expressed higher HDAC11 levels. Bevacizumab-treated (1×) cells had significant change in HDAC1, HDAC6, and HDAC11. In cultures treated with 2× bevacizumab, only HDAC11 expression levels were significantly affected compared with controls. Aflibercept-treated (1×) cells had changes in expression of HDAC1, HDAC6, and HDAC11. At 2× concentration, only HDAC11 was significantly changed. CONCLUSION: Our results show that antiangiogenic drugs can affect the transcription profile of genes regulating the histone acetylation status in ARPE-19 cells in vitro. This finding may have an implication in differential patient response to anti-vascular endothelial growth factor therapy by means of possible interactions between treatment and patient's epigenomic profile. [Ophthalmic Surg Lasers Imaging Retina. 2018;49:S29-S33.].

Protective Effects of Memantine on Hydroquinone-Treated Human Retinal Pigment Epithelium Cells and Human Retinal Müller Cells.

PURPOSE: Memantine (MEM) acts on the glutamatergic system by blocking N-methyl-d-aspartate (NMDA) glutamate receptors. The role that MEM plays in protecting retinal cells is unknown. Hydroquinone (HQ) is one of the cytotoxic components in cigarette smoke. In the present study, we tested whether pretreatment with MEM could protect against the cytotoxic effects of HQ on human retinal pigment epithelium cells (ARPE-19) and human retinal Müller cells (MIO-M1) in vitro. METHODS: Cells were plated, pretreated for 6 h with 30 µM of MEM, and then exposed for 24 h to 200, 100, 50, and 25 µM of HQ while MEM was still present. Cell viability (CV), reactive oxygen species (ROS), mitochondrial membrane potential (ΔΨm), and lactate dehydrogenase (LDH) release assays were performed. RESULTS: HQ-treated cells showed a dose-dependent decrease in CV and ΔΨm, but an increase in ROS production and LDH levels in both cell lines. MEM pretreatment reversed the CV in 50, 100, and 200 µM doses in ARPE-19 cells and at all HQ concentrations in MIO-M1 cells compared to HQ-treated cultures. ROS production was reversed in all HQ concentrations in both cell lines. ΔΨm was significantly increased after MEM pretreatment only in 50 µM HQ concentration for both cell lines. LDH levels were decreased at 50 and 25 µM HQ in ARPE-19 and MIO-M1 cells, respectively. CONCLUSION: HQ-induced toxicity is concentration dependent in ARPE-19 and MIO-M1 cultures. MEM exerts protective effects against HQ-induced toxicity on human retinal pigment epithelial and Müller cells in vitro.