July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Role of nuclear factor activated in T cells (NFAT) signaling pathway in the Retinal Pigment Epithelial cells
Author Affiliations & Notes
  • Hsuan-Yeh Pan
    optometry, Indiana University, Bloomington, Indiana, United States
  • Mallika Valapala
    optometry, Indiana University, Bloomington, Indiana, United States
  • Footnotes
    Commercial Relationships   Hsuan-Yeh Pan, None; Mallika Valapala, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 1941. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Hsuan-Yeh Pan, Mallika Valapala; Role of nuclear factor activated in T cells (NFAT) signaling pathway in the Retinal Pigment Epithelial cells. Invest. Ophthalmol. Vis. Sci. 2019;60(9):1941.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : The nuclear factor activated in T cells (NFAT) is a family of transpiration factors that play an important role in integrating Ca2+ signaling with a variety of inflammatory processes. Of the five identified NFAT family members (NFAT1-5), four isoforms NFAT1-4 are subject to dephosphorylation and activation by calcineurin. Dephosphorylation by calcineurin results in NFAT nuclear translocation and transcriptional induction of NFAT-regulated genes, including, interleukin (IL)-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-10, IL-13, IFN-γ, TNF-α, and granulocyte-macrophage colony-stimulating factor (GM-CSF). We investigated the role of NFAT proteins in activating transcription of inflammatory cytokines in the RPE.

Methods : Transcriptional induction of NFAT was measured using the adenoviral NFAT reporter vector (AD-NFAT-luc). NFAT transcriptional activity was inhibited by a peptide, MAGPHPVIVITGPHEE (VIVIT), which known to selectively prevent the interaction of NFAT with calcineurin and inhibit nuclear translocation of NFAT. Quantitative Real time-PCR (qRT-PCR) analysis was used to determine the expression of cytokines: Interleukin 6 (IL-6), Interleukin 8 (IL-8)

Results : ARPE-19 cells transfected with the AD-NFAT-luc vector for 24 hrs followed by treatment with 10 µg/ml LPS for 24 hrs showed a significant induction in luciferase activity. LPS-induced NFAT luciferase activity was reduced by treatment of cells with the NFAT inhibitor, VIVIT for 6 hrs. qRT-PCR analysis revealed increased expression of IL-6 and IL-8 in cells treated with LPS compared to untreated cells. LPS-induced expression of IL-6 and IL-8 was inhibited by treatment with the NFAT inhibitor,VIVIT.

Conclusions : These results suggest an important role of NFAT in regulating the expression of IL-6 and IL-8 in the RPE. Furthermore, inhibition of NFAT decreases LPS- induced expression of inflammatory cytokines in the RPE.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×