Abstract: FR-PO457
Plasminogen Activator Inhibitor 1 (PAI-1) and Nuclear Factor of Activated T-Cells 5 (NFAT5) Mediates TGFβ-Induced Phenotype Transition of Human Peritoneal Mesothelial Cells (MCs)
Session Information
- Peritoneal Dialysis: Current Topics
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Dialysis
- 702 Dialysis: Home Dialysis and Peritoneal Dialysis
Authors
- Kim, Dal-Ah, Ewha Womans University, Seoul, Korea (the Republic of)
- Kang, Hyun-Jung, Ewha Womans University, Seoul, Korea (the Republic of)
- Her, Ye rim, Ewha Womans University, Seoul, Korea (the Republic of)
- Kang, Duk-Hee, Ewha Womans University, Seoul, Korea (the Republic of)
Background
The epithelial-to-mesenchymal transition (EMT) of MCs is an early mechanism of peritoneal dysfunction in peritoneal dialysis (PD). Plasminogen activator inhibitor-1 (PAI-1) is an inhibitor of fibrinolysis by hindering the proteolytic activity of tissue type plasminogen activator, and recently reported to regulate EMT of cancer cells. NFAT5 is known to regulate PAI-1 transcription, and also to promote EMT of cancer cells. The aim of this study is to investigate the role of PAI-1 and NFAT5 in TGFβ-induced EMT of MCs and its mechanism.
Methods
EMT was evaluated by the changes in morphology and markers of epithelial and mesenchymal cells. Effect of gene silencing of PAI-1 or PAI-1 inhibitor (Tiplaxtinin, 20 uM) on TGFβ-induced EMT was examined. Activation of Smad2/3, Erk1/2 MAPK phosphorylation, snail expression, nuclear translocation of snail was assessed. MMP2 expression was evaluated by real time PCR and WB. The interaction between NFAT5 and β-catenin was analyzed by immunoprecipitation. RNA-seq was performed to analyze gene profiling in MCs isolated from omentum (HPMC, N=4) or MCs isolated from overnight dwell dialysates from PD patients (PDMC, N=9), respectively.
Results
Either PAI-1 gene silencing or Tiplaxtinin ameliorated TGFβ-induced changes in cell morphology and the expression of E-cadherin, α-SMA, and fibronectin. TGFβ-induced decrease in E-cadherin promoter activity and nuclear translocation of snail were also alleviated by siPAI-1. PAI-1 gene silencing ameliorated TGFβ-induced activation of smad2/3 and Erk1/2 MAPK in HPMC. In addition, TGFβ-induced increase MMP2 expression was alleviated by siPAI-1. TGFβ-induced increase in PAI-1 was blocked by siNFAT5. siPAI-1 also ameliorated TGFβ-induced NFAT5/β-catenin interaction. RNA-seq analysis revealed an increased in PAI-1 expression in PDMC compared to HPMC, which was associated with altered expression of EMT-related genes.
Conclusion
PAI-1 plays a role in TGFβ-induced EMT of peritoneal MCs via an interaction with NFAT5, and modulation of PAI-1 expression/activity in MCs could be a novel strategy to prevent peritoneal fibrosis in PD patients.