Abstract
Purpose :
Endoplasmic reticulum (ER) stress-mediated cell death contributes to many ocular diseases, including Fuchs corneal endothelial dystrophy (FECD). Natural compounds possess significant pharmacological activities necessary for vital cell signaling pathways and are considered protective in many ocular diseases such as diabetic retinopathy and glaucoma. However, there are no studies understanding the role of natural compounds in reducing sustained ER stress for improving corneal endothelial cell (CEnCs) survival. The purpose of this study is to identify readily available natural compounds to improve the viability of corneal endothelial cells (CEnC) under ER stress.
Methods :
Human corneal endothelial cells (21T-HCEnC) were cultured and seeded in 384-well plates, pre-treated with 404 compounds from a natural product library (Enzo Screen-Well Natural Product library), and then subjected to ER stressor tunicamycin (1µg/mL) for 24 hours. Cell viability was measured using Cell-Titer Glo assay, and compounds improving viability past the set threshold were selected for further dose-response testing.
Results :
Assay standardization was performed prior to compound screening, where plate density, compound, and stressor concentrations were standardized. Forty-one compounds of the 404 natural products screened, each administered at 0.01 µg/mL, exhibited a marked increase in cell viability compared to tunicamycin-treated cells. Of the forty-one compounds showing an increase, 33,7 and 1 compounds showed 30%-40%, 40%-50%, and > 50% increase in viability, respectively, compared to tunicamycin-treated cells. Most of these compounds fell under the classification of alkaloids or antibiotics, with forskolin and naringenin being two products seen with the highest percentage of CEnCs protection.
Conclusions :
The proposed study will provide insight into the role of natural compounds in regulating ER stress-mediated CEnC death associated with many corneal endothelial-associated diseases. Future studies are needed to identify more selective ‘hits’ and to clarify the mechanisms of action in regulating ER stress.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.