June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Extracellular matrix nitration alters growth factor release and activates bioactive complement in human retinal pigment epithelium
Author Affiliations & Notes
  • Mark Fields
    Ophthalmology and Visual Science, Yale School of Medicine, New Haven, Connecticut, United States
  • Hannah Bowrey
    Rutgers Brain Health Institute, The State University of New Jersey, Piscataway, New Jersey, United States
  • Jie Gong
    Ophthalmology and Visual Science, Yale School of Medicine, New Haven, Connecticut, United States
  • Ernesto F Moreira
    Ophthalmology, Medical University of South Carolina, Charleston, South Carolina, United States
  • Hui Cai
    Ophthalmology and Visual Science, Yale School of Medicine, New Haven, Connecticut, United States
  • Lucian Del Priore
    Ophthalmology and Visual Science, Yale School of Medicine, New Haven, Connecticut, United States
  • Footnotes
    Commercial Relationships   Mark Fields, None; Hannah Bowrey, None; Jie Gong, None; Ernesto Moreira, None; Hui Cai, None; Lucian Del Priore, None
  • Footnotes
    Support  Research to Prevent Blindness; Foundation Fighting Blindness
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 2265. doi:
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    • Get Citation

      Mark Fields, Hannah Bowrey, Jie Gong, Ernesto F Moreira, Hui Cai, Lucian Del Priore; Extracellular matrix nitration alters growth factor release and activates bioactive complement in human retinal pigment epithelium
      . Invest. Ophthalmol. Vis. Sci. 2017;58(8):2265.

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

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Abstract

Purpose : We have shown previously that non-enzymatic nitration (NEN) of extracellular matrix, which serves as a model Bruch’s membrane (BM) aging, has a profound effect on the behavior of retinal pigment epithelium (RPE), including altered phagocytic ability, reduced cell adhesion, and inhibition of proliferation. We know that transplanted RPE will encountered a hostile subretinal environment, including BM alterations that may compromise cell function and survival. Here we use our previous model of BM aging (NEN) to determine the effects of NEN on growth factor release and complement activation in RPE.

Methods : Human induced pluripotent stem cells (iPSC) were differentiated into RPE, and analyzed by immunohistochemistry, confocal microscopy, and polymerase chain reaction (PCR). IPSC-derived RPE were plated onto RPE-derived extracellular matrix (ECM) conditions (untreated or nitrite-modified). Cells were cultured for 7 days and barrier function measured by transepithelial resistance (TER) and vascular endothelial growth factor (VEGF), pigment epithelium-derived factor (PEDF), and C3a were measured using enzyme linked immunosorbent assay (ELISA).

Results : Nitrite-modified ECM increased VEGF release both apically and basally by 0.14 ng/mL (p <0.001) and 0.27 ng/mL (p <0.001), respectively, in iPSC-derived RPE. Nitrite-modified ECM increased PEDF release in iPSC-derived RPE apically by 0.26 ng/mL (p <0.001). Nitrite-modified ECM increased production of C3a both apically and basally in iPSC-derived RPE by 0.19 ng/mL (p <0.001) and 0.15 ng/mL (p <0.001), respectively.

Conclusions :
Nitrite-modified ECM increase VEGF, PEDF release, and C3a production in human iPSC-derived RPE. This model demonstrates changes seen in the pathophysiology of age-related macular degeneration.

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

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