Expression and activation of MMP10 in retinal pigment epithelium cell lines and plasma levels in age‐related macular degeneration patients

Abstract

AbstractPurpose: The aim of the study is to evaluate the role of MMP10 in AMD at the cellular level and in human samples.Methods: To determine expression in retinal pigment epithelium (RPE) cells, immunofluorescence labeling and western blotting (WB) and relative quantification PCR (RQ‐PCR) were performed in ARPE‐19 and human RPE primary cells (hRPE). To evaluate the effect of oxidative stress conditions on MMP10 expression and activation, cells were subjected cells exposed to H2O2 at 800 μM during 48 h. To evaluate the levels of MMP10 in AMD patients, the plasma concentration measured by ELISA in an age‐ and sex‐matched 1:1 case–control cohort (104 patients).Results: MMP10 is expressed in ARPE‐19 cells and hRPE cells under basal conditions. ARPE‐19 cells exposed to showed a prominent increase in MMP10 expression in the immunofluorescence labeling. WB analysis of MMP10 under basal conditions in the supernatant and cell lysate of ARPE‐19 and hRPE cells have a marked expression of MMP10 contained intracellularly and present a measurable concentration of extracellular MMP10. H2O2 exposition seems to activate the protein generating two visible bands in WB. RQ‐PCR shows a significant increase in MMP10 mRNA transcription over time under oxidative stress conditions. Comparison of mean MMP10 plasma showed no statistically significant differences. A statistically significant negative correlation between age and MMP10 plasma levels was observed in the control while no apparent correlation was found in the AMD group.Conclusions: Our study demonstrates that MMP10 is expressed in human epithelial cells and is overexpressed and activated under oxidative stress conditions. This increase in expression by RPE observed in cells does not correlate with an increase in plasma concentration in patients with AMD, however, in contrast to what is observed in controls, there is no decrease in its systemic concentration with age as opposed to what occurs in control patients. The data suggest that MMP10 may play a role in the development of AMD and future experiments are needed to explore this possibility.