The effect of corticosteroids on human choroidal endothelial cells: a model to study central serous chorioretinopathy

Abstract

Purpose : Central serous chorioretinopathy is a relatively common chorioretinal disease that can lead to significant vision loss. Endogenous and exogenous hypercortisolism are the most important risk factors for central serous chorioretinopathy. The disease appears to originate in the choroid, with secondary breakdown of the retinal pigment epithelial outer blood-retinal barrier, resulting in subretinal fluid accumulation. However, the exact pathogenesis is unknown. We developed a model to allow for the investigation of corticosteroid administration to primary human choroidal endothelial cells, to unravel the role of corticosteroids in central serous chorioretinopathy.

Methods : Choroidal endothelial cells were isolated from cadaveric human donors. Magnetic-activated cell sorting with anti-human CD31 was performed for choroidal endothelial cell isolation. Isolated choroidal endothelial cell cultures had a cobblestone appearance in monolayer cultures and stained positive for the cell-cell adhesion glycoprotein cadherin 5 (vascular endothelial cadherin). Moreover, on 3D Matrigel matrix these cells were able to form capillary-like structures, characteristic of in vitro endothelial cell function. Primary cultures of purified choroidal endothelial cells treated with intermittent administration of 10^-7 M cortisol were analysed for the effect on both direct and downstream putative corticosteroid responsive genes (FKBP5, PER1, GILZ1, and SGK1).

Results : We found that intermittent administration of 10^-7 M cortisol (mimicking the in vivo situation with diurnal rhythm in blood cortisol levels) led to significant transcriptional upregulation of validated cortisol target genes. Further pharmacological analysis identified the glucocorticoid receptor rather than the mineralocorticoid receptor as the mediator of the cortisol effect on gene expression.

Conclusions : With this optimized choroidal endothelial cell isolation and culturing protocol, we have established an in vitro model that appears very suitable for research on both central serous chorioretinopathy and other diseases in which corticosteroids and choroidal endothelial cells are involved.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.