July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Dysregulation of MMP2/MMP9 activity by both local (RPE, vasculature) and systemic (serum) factors promotes macular-degeneration-relevant pathological changes in iPSC-derived model(s) of the disease.
Author Affiliations & Notes
  • Kannan Vrindavan Manian
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Sonal Dalvi
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Chad A Galloway
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Lauren Winschel
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Arushi Jain
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Celia Soto
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Ruchira Singh
    Opthalmology, University of Rochester Medical Center, Rochester, New York, United States
  • Footnotes
    Commercial Relationships   Kannan Manian, None; Sonal Dalvi, None; Chad Galloway, None; Lauren Winschel, None; Arushi Jain, None; Celia Soto, None; Ruchira Singh, None
  • Footnotes
    Support  BrightFocus Foundation Macular Degeneration Grant, Foundation of Fighting Blindness Individual Investigator Award, National Institute of Health, NIH-1R01EY028167, and Research to Prevent Blindness, RPB’s Career Development Award, RPB's Unrestricted Challenge Grant to Department of Ophthalmology at University of Rochester
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 1227. doi:
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      Kannan Vrindavan Manian, Sonal Dalvi, Chad A Galloway, Lauren Winschel, Arushi Jain, Celia Soto, Ruchira Singh; Dysregulation of MMP2/MMP9 activity by both local (RPE, vasculature) and systemic (serum) factors promotes macular-degeneration-relevant pathological changes in iPSC-derived model(s) of the disease.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):1227.

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

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Abstract

Purpose : Impaired extracellular matrix (ECM) turnover due to matrix metalloproteinase(s) (MMPs) dysfunction is implicated in AMD and related macular dystrophies (MDs). However, the underlying basis of MMP dysfunction in AMD/MDs has not been established. Our goal in this study was to utilize iPSC-derived AMD/MD model(s) to i) investigate the impact of local (RPE and vasculature) and systemic (serum) influences on the expression and activity of specific MMPs and ii) determine its consequence for development of drusen, ECM protein accumulation and neovascularization.

Methods : hiPSC-derived cell model(s) of AMD/MDs were established using iPSC-RPE, iPSC-endothelial cells (ECs) and iPSC-EC-derived vascular network and physiologically relevant stressors, serum from either control subjects or patients with AMD. Drusen formation, ECM protein accumulation and neovascularization phenotypes were assessed by immunocytochemistry, Western blotting, cell proliferation, ECM-invasion, tube formation and branching assays. Furthermore, the expression, localization and/or activity of specific MMPs, MMP2, MMP9 and MMP14, was assessed by Western blotting, immunocytochemistry, gel zymography and in-situ zymography.

Results : MMP2 and MMP9 co-localized with known drusen-resident proteins (e.g. APOE) in drusen deposits beneath AMD/MD iPSC-RPE cells. Furthermore, in the presence of normal serum, MMP2 and MMP9 activity was lower in RPE conditioned media (RPE-CM) from AMD/MD iPSC-RPE cultures compared to RPE-CM from control iPSC-RPE cultures. Consistent with this finding, higher amounts of COL4, an ECM protein whose turnover is regulated by MMP2 and MMP9, was seen in ECM underneath AMD/MD iPSC-RPE cultures compared to ECM underneath control iPSC-RPE cultures. With respect to neovascularization, AMD/MD iPSC-RPE-CM and AMD serum either directly enhanced the activity of MMP2 and MMP9 or led to increased levels of FGF2, a pro-angiogenic molecule that modulates MMP2 and MMP9 activity, and thereby promoted both ECM invasion by hiPSC-ECs and vascular density in hiPSC-EC-derived vascular network cultures.

Conclusions : Using iPSC model(s) of AMD/MDs, we show that both local (RPE and vascular networks) and systemic (serum) factors can alter MMP2 and MMP9 activiity and promote drusen formation, ECM protein accumulation and neovascularization.

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

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