June 2020
Volume 61, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2020
Cell-autonomous RPE dysfunction is sufficient to instigate CC atrophy and CNV in an iPSC model of Macular Degeneration
Author Affiliations & Notes
  • Kannan Vrindavan Manian
    1. Department of Ophthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
  • Sonal Dalvi
    1. Department of Ophthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
  • Chad A Galloway
    1. Department of Ophthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
  • Celia Soto
    1. Department of Ophthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
  • Lauren Winschel
    1. Department of Ophthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
  • Yiming Li
    Department of Biomedical Engineering, University of Rochester, Rochester, New York, United States
  • Danielle S W Benoit
    Department of Biomedical Engineering, University of Rochester, Rochester, New York, United States
  • Ruchira Singh
    1. Department of Ophthalmology and Biomedical Genetics, University of Rochester, Rochester, New York, United States
    UR Stem Cell and Regenerative Medicine Center, University of Rochester, Rochester, New York, United States
  • Footnotes
    Commercial Relationships   Kannan Manian, None; Sonal Dalvi, None; Chad Galloway, None; Celia Soto, None; Lauren Winschel, None; Yiming Li, None; Danielle Benoit, None; Ruchira Singh, None
  • Footnotes
    Support  BrightFocus Foundation Macular Degeneration Grant, David Bryant Trust,Foundation of Righting Blindness Individual Investigator Award, Knights Templar eye foundation, NIH­1R01EY028167, NIH­1R01 EY030183, RPB's Career Development Award
Investigative Ophthalmology & Visual Science June 2020, Vol.61, 3809. doi:
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    • Get Citation

      Kannan Vrindavan Manian, Sonal Dalvi, Chad A Galloway, Celia Soto, Lauren Winschel, Yiming Li, Danielle S W Benoit, Ruchira Singh; Cell-autonomous RPE dysfunction is sufficient to instigate CC atrophy and CNV in an iPSC model of Macular Degeneration. Invest. Ophthalmol. Vis. Sci. 2020;61(7):3809.

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

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Abstract

Purpose : The purpose of this study was to use a representative hydrogel-based model of the RPE-CC complex to investigate RPE-choriocapillaris (CC) cross-talk in the development of CC atrophy and choroidal neovascularization (CNV), disease hallmarks of age-related macular degeneration (AMD) and related macular dystrophies, like Sorsby’s fundus dystrophy (SFD).

Methods : SFD (TIMP3, S204C) and control induced pluripotent stem cells (iPSCs) were differentiated into RPE, endothelial cells (ECs) and mesenchymal stem cells (MSCs). To recapitulate in-vivo RPE-CC structure, hiPSC-ECs were encapsulated in PEG hydrogels plated on a layer of hiPSC-MSCs and hiPSC-RPE was grown on the vascularized gel surface. Immunostaining and Western blotting analyses were performed to characterize RPE and CC characteristics in the hydrogel-based control vs. SFD RPE-CC tissue mimetic. Furthermore, Alamar blue cell proliferation assay, extracellular matrix (ECM)-cell invasion and migration assay and immunocytochemical analyses of vascular network characteristics (number, size, length) were used to evaluate the effect of control vs. SFD RPE-secreted factors on CC atrophy and CNV-relevant changes. All experiments were performed at least in triplicates and unpaired t-test was utilized to determine statistical significance (p<0.05).

Results : Characterization of hiPSC-RPE-CC model showed an RPE monolayer that expressed tight junction marker ZO-1 overlying a CC-like fenestrated endothelium that showed co-expression of EC-specific protein, CD31 and fenestration marker, PLVAP. Furthermore, consistent with CC atrophy and CNV in SFD, SFD-hiPSC-RPE-CC model showed vascular EC atrophy and invasion of RPE by ECs. Furthermore, SFD-hiPSC-RPE conditioned media alone was sufficient to induce CC atrophy and CNV relevant phenotypes (ECM invasion and migration (p< 0.05), EC proliferation(p< 0.05)) in ECs/CC-like vascular networks. Notably, pharmacological inhibition of FGF2 was sufficient to suppress CNV relevant phenotypes in this SFD hiPSC model (p< 0.01).

Conclusions : Using an iPSC-RPE-CC model, we have shown that alterations in RPE-secreted factors are sufficient to cause vascular changes, CC atrophy and CNV, observed in SFD/AMD. Furthermore, iPSC-RPE-CC model can be used to pharmacologically-target SFD/AMD relevant pathological changes, like CNV

This is a 2020 ARVO Annual Meeting abstract.

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