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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)
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).
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).
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).
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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