A novel accessible in vitro model of proliferative vitreoretinal diseases shows facilitated epithelial mesenchymal transition through aquaporin-1

This study aimed to establish a new in vitro model of proliferative vitreoretinal disease (PVD) using ARPE-19 cells and to evaluate the role of Aquaporin-1 (AQP1) in this model. ARPE-19 cells, stably transfected without or with AQP1, were differentiated in a nicotinamide-enriched medium (N + medium) prior to a 5 day treatment with either N + medium, medium devoid of nicotinamide (N-), or N- medium supplemented with TNF-α and TGF-β (TNT) to induce epithelial-mesenchymal transition (EMT). Human proliferative vitreoretinal disease (PVD) membranes and retinal pigmented cells isolated from ocular globes were collected. RNA sequencing was performed on both ARPE-19 treated cells and human samples. Cell migration assays and immunofluorescence analyses were performed to assess the effects of cell treatments. After differentiation in N + medium, control and AQP1-transfected ARPE-19 cells treated with TNT in N- medium (CT TNT N- and AQP1 TNT N-) exhibited features of EMT, including elongated spindle-like morphology and enhanced migratory capacity. AQP1 TNT N- cells shared the highest transcriptional similarities with human PVD membranes. Moreover, in the absence of TNT, AQP1-transfected ARPE-19 cells in N- medium exhibited increased migration and early EMT markers, suggesting a role for AQP1 in these processes. Importantly, AQP1 expression appeared to slightly upregulate matrix metallopeptidase 2 (MMP2) expression, which may promote cell invasion. Zinc finger E-box-binding homeobox-1 (ZEB1), a key EMT regulator, was upregulated in conditions containing TNT and AQP1. Our findings suggest that the AQP1 TNT N- model best mimics human PVD membranes, both in transcriptomic profile and cellular behavior.