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<p>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&#xa0;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 (<i>MMP2</i>) expression, which may promote cell invasion. Zinc finger E-box-binding homeobox-1 (<i>ZEB1</i>), 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.</p>

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

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年12月24日

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  • Azine Datlibagi,
  • Clément Duveau,
  • Anna Zein-El-Din,
  • Nargis Bolaky,
  • Vincent Lambert,
  • Jason Perret,
  • François Willermain,
  • Christine Delporte,
  • Elie Motulsky

摘要

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.