June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
NFAT-dependency of IL-1β-induced diabetes-relevant behaviors in human retinal microvascular endothelial cells and Müller cells
Author Affiliations & Notes
  • Meredith J Giblin
    Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee, United States
  • Megan E Capozzi
    Molecular Physiology & Biophysics, Vanderbilt University, Nashville, Tennessee, United States
  • Gary W. McCollum
    Department of Ophthalmology and Visual Sciences, Vanderbilt University, Nashville, Tennessee, United States
  • John S Penn
    Department of Ophthalmology and Visual Sciences, Vanderbilt University, Nashville, Tennessee, United States
    Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee, United States
  • Footnotes
    Commercial Relationships   Meredith Giblin, None; Megan Capozzi, None; Gary McCollum, None; John Penn, None
  • Footnotes
    Support  T32EY021453, R01-EY007533, R01-EY023639, and P30-EY008126; Research To Prevent Blindness and The Reeves Foundation.
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 4044. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to Subscribers Only
      Sign In or Create an Account ×
    • Get Citation

      Meredith J Giblin, Megan E Capozzi, Gary W. McCollum, John S Penn; NFAT-dependency of IL-1β-induced diabetes-relevant behaviors in human retinal microvascular endothelial cells and Müller cells. Invest. Ophthalmol. Vis. Sci. 2017;58(8):4044.

      Download citation file:


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

      ×
  • Supplements
Abstract

Purpose : Early diabetic retinopathy (DR) involves chronic low-grade inflammation, characterized by elevated vitreous cytokines, such as IL-1β. This inflammation promotes many pathologic consequences, including the hallmark vascuar changes for which DR is best known. Nuclear factor of activated T cells (NFAT) is involved in the regulation of inflammatory mediators, extracellular matrix proteins, and adhesion molecules and thus may control multiple pathogenic steps early in DR. We have examined NFAT's role in IL-1β auto-amplification and explored the NFAT-dependency of endothelial cell responses to IL-1β, including monolayer permeability and expression of targets involved in leukocyte adhesion and basement membrane thickening.

Methods : For IL-1β production, human Müller cells (HMC) were treated with 50pg/mL IL-1β, Inhibitor of NFAT-Calcineurin Association-6 (INCA, 2.5μM) and proper vehicles for 8 hrs before collection for qRT-PCR. For ICAM expression, human retinal microvascular endothelial cells (HRMEC) were treated with 1ng/mL IL-1β and 2.5μM INCA for 2 hrs. For collagen IV expression, HRMEC were treated with 10ng/mL IL-1β and 1μM INCA for 48 hrs. In permeability experiments, HRMEC were pre-treated with vehicle or 2.5μM INCA for 16 hrs before 24 hrs of exposure to 0.5ng/mL IL-1β with/out INCA. Transendothelial electrical resistance values were measured using the EVom2.

Results : IL-1β increased HMC IL-1β expression by 82-fold (p<0.01); INCA inhibited this induction by 28% (p<0.05). HRMEC treatment with IL-1β caused a 328-fold induction of ICAM expression (p<0.01); INCA inhibited this induction by 20% (p<0.01). HRMEC treatment with IL-1β caused a 2-fold induction of collagen IV expression (p<0.01); INCA inhibited this induction by 50% (p<0.01). Treatment with IL-1β decreased HRMEC monolayer resistance by 25% (measured vs resistance at time=0, p<0.01), which was partially rescued (44%, p<0.01) by INCA treatment.

Conclusions : These data demonstrate the potential of NFAT as a multi-targeted therapy for retinal inflammation secondary to diabetes. NFAT inhibition not only prevented HMC IL-1β auto-amplification, but also inhibited the pathogenic response of HRMEC to IL-1β, including the expression of adhesion proteins, excessive extracellular matrix deposition and increased permeability. Future work will investigate the therapeutic potential of in vivo NFAT inhibition.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

×
×

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.

×