Kidney Week

Abstract: SA-PO932

Novel Role of Nuclear Factor of Activated T-Cells 5 (NFAT5) as a Mediator of EMT of Peritoneal Mesothelial Cells (MCs) via Modulation of Nod-Like Receptor-3 (NLRP3) Inflammasome

Session Information

• Dialysis: Peritoneal Dialysis - III
  October 27, 2018 | Location: Exhibit Hall, San Diego Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Dialysis

• 703 Dialysis: Peritoneal Dialysis

Authors

• Kang, Duk-Hee, Ewha University College of Medicine, Seoul, Korea (the Republic of)

Duk-Hee Kang,

• Ryu, Eun sun, Ewha Womans University School of Medicine, Seoul, Korea (the Republic of)

Eun sun Ryu,

• Kim, Dal-ah, Ewha Womans University Medical Center, Seoul, Seoul, Korea (the Republic of)

Dal-ah Kim,

• Kang, Hyun-Jung, Ewha Womans University, Seoul, Korea (the Republic of)

Hyun-Jung Kang,

• Kim, You Jin, Kyungpook National University, Daegu, Korea (the Republic of)

You Jin Kim,

• Park, Sun-Hee, Kyungpook National University Hospital, Daegu, Korea (the Republic of)

Sun-Hee Park,

• Kim, Yong-Lim, Kyungpook National University Hospital, Daegu, Korea (the Republic of)

Yong-Lim Kim,

• Kwon, Hyug Moo, UNIST, Ulsan, Korea (the Republic of)

Hyug Moo Kwon,

Background

Epithelial-to-mesenchymal Transition (EMT) of MCs is known as an early mechanism of peritoneal fibrosis in PD. NLRP3 inflammasome is comprised of the NLRP3, ASC and procaspase-1, which leads to the maturation of IL-1β and IL-18. NFAT5 is an essential transcription factor regulating cellular homeostasis to hypertonicity-induced osmotic stress, and recently reported as a pro-inflammatory mediator. We investigated whether NFAT5 plays a role in peritoneal EMT via a modulation of NLRP3 inflammasome in MCs.

Methods

The expressions of NFAT5, components of NLRP3 inflammasome, nuclear translocation of NFAT5, β-catenin and snail were evaluated by western blotting. EMT was evaluated by the changes in morphology and markers of epithelial and mesenchymal cells. E-cadherin promoter activity was confirmed by luciferase assay. Effect of gene silencing of NFAT5, NLRP3 and ASC on EMT was examined using siRNA. Animal model of peritoneal fibrosis was established by intraperitoneal injection of adenoviral vector of TGFβ in BL6 mouse.

Results

TGFβ increased NFAT5 expression and its nuclear translocation in MC. TGFβ-induced EMT was associated with an up-regulation of NLRP3, ASC, procaspase-1 and an increased production of IL-1β/IL-18. In adenoviral TGFβ-injected mice, EMT of MC with submesothelial fibrosis was observed with an increased expression of NFAT5 and NLRP3. siNFAT5 ameliorated TGFβ-induced EMT with an increase in E-cadherin promotor activity as well as a decrease in snail expression and nuclear translocation of β-catenin. siNLRP3, siASC and neutralizing antibodies of IL-1β and IL-18 alleviated TGFβ1-induced EMT. siNFAT5 also alleviated TGFβ1-induced activation of NLRP3 inflammasome. Neither MAPK inhibitors (PD98059, SB203580) nor antioxidants (NAC, Apocynin, Mitotempo) altered TGFβ1-induced up-regulation of NFAT5 of MC.

Conclusion

This data suggest NFAT5 plays a key role in peritoneal EMT via tonicity-independent mechanism by either an inhibition of E-cadherin transcription and activation of NLRP3 inflammasome. Modulation of NFAT5 and NLRP3 inflammasome in MCs could be a novel approach to protect the peritoneum from the development of EMT and peritoneal fibrosis in PD patients.