July 2019
Volume 60, Issue 9
ARVO Annual Meeting Abstract  |   July 2019
The Influence of Corticosteroids on Choroidal Endothelial Cells in Central Serous Chorioretinopathy
Author Affiliations & Notes
  • Rebecca Kaye
    Ophthalmology, University of Southampton, Winchester, United Kingdom
  • David R. G. Christensen
    Ophthalmology, University of Southampton, Winchester, United Kingdom
  • Andrew Lotery
    Ophthalmology, University of Southampton, Winchester, United Kingdom
  • Footnotes
    Commercial Relationships   Rebecca Kaye, None; David Christensen, None; Andrew Lotery, None
  • Footnotes
    Support  Gift of Sight Eye Research Charity
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 4326. doi:
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      Rebecca Kaye, David R. G. Christensen, Andrew Lotery; The Influence of Corticosteroids on Choroidal Endothelial Cells in Central Serous Chorioretinopathy. Invest. Ophthalmol. Vis. Sci. 2019;60(9):4326.

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

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Purpose : The choroid is acknowledged to be the initial site of pathology in central serous chorioretinopathy (CSCR) and corticosteroids a significant risk factor for disease. Choroidal tissue from CSCR donors is rare. We describe the use of induced pluripotent stem cells (hiPSCs) differentiated into choroidal endothelial cells (CECs) from chronic CSCR donors to study the response to steroids in vitro.

Methods : Skin fibroblasts cultured from a CSCR donor were induced to pluripotency using Sendai virus. hiPSCs were differentiated into CECs using a co-culture technique with the primate CEC line RF6A and sorted for the endothelial cell marker CD144. hiPSC-derived CECs were characterised using immunocytochemistry. Functional assays were performed including trans-endothelial electrical resistance (TEER) and tube-formation assays. The response of hiPSC-derived CECs to corticosteroids was compared to RF6A cells using TEER, tube-formation assays and western blotting. Tube-formation & western blotting results normalised to control cultures. Statistical analysis using student’s t-test.

Results : Karyotypically normal hiPSC-derived CECs from a chronic CSCR donor showed significantly reduced TEER (1.87±3.33Ω*cm2, p<0.05) after addition of aldosteronex10-5M at 24hrs in comparison to control cultures (24.76±2.83Ω*cm2). Tube-formation was significantly reduced (0.38±0.22 tubes relative to control, p<0.05) after addition of aldosteronex10-5M at 16hrs. RF6A were not similarly affected. Addition of cortisol at x10-5–x10-7M and aldosterone at x10-5–x10-9M to hiPSC-derived CECs showed significantly reduced ratio of phosphorylated extracellular regulated kinase (pERK) to ERK relative to control cultures on western-blotting (p<0.001). Addition of aldosterone x10-5M and cortisol x10-6M reduced pERK:ERK in RF6A cells relative to control (p<0.005).

Conclusions : Characteristically normal hiPSC-derived CECs can be produced from chronic CSCR donors. Addition of corticosteroids, specifically aldosterone significantly reduced TEER and tube-formation. Mitogen-activated protein kinase enzyme inhibitor related retinopathy, in which pERK is chemically inhibited gives a similar clinical picture to CSCR. Interestingly pERK:ERK was significantly reduced on addition of corticosteroids in hiPSC-derived CECs; possibly hinting at a common molecular pathway for CSCR. These results suggest the function of CECs are affected by corticosteroids.

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


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