Abstract
Purpose :
The goal of this project was to analyze the relationship between cell morphology and proteases/proteases inhibitors (PIs) secretion profile in Fuchs Endothelial Corneal Dystrophy (FECD) corneal endothelial cells (CECs).
Methods :
Cell morphology was determined using a circularity index (4π*area/perimeter2) for each CEC population extracted from surgical FECD specimens (N=2) and healthy Eye bank corneas (N=3). CECs were cultured 28 days post-confluency. Supernatant was collected and analysed using Proteome Profiler Array detecting 35 proteases and 32 PIs (R&D Systems). Proteome signal was analyzed using Image Studio Lite and correlated with the population’s circularity index.
Results :
Calculation of circularity index reported different morphologies among FECD populations (0,59±0,18 and 0,64±0,17) and healthy populations (0,44±0,18, 0,66±0,13 and 0,71±0,11). Proteome arrays revealed the presence of 10 proteases (ADAMTS1, Cathepsin A, B, D, and X/Z/P, DPPIV/CD26, MMP-2, 3 and 12, uPA/Urokinase) and 10 PIs (Protease Nexin II, Cystatin B and C, EMMPRIN/CD147, Latexin, Lipocalin-1, Serpin E1, TFPI, TFPI-2, TIMP-1, 2 and 4). Healthy and FECD specimens showed similar variation patterns according to morphology for secretion of ADAMTS1, MMP-3 and 12. However, opposing patterns between healthy and FECD populations were observed for Cathepsin B and D. Moreover, some proteins did not show variation according to phenotype in healthy CECs, but did in FECD CECs: Cathepsin A, Cystatin C, TFPI-2 and total TIMPs. For the other proteins, secretion did not vary according to morphology or no specific pattern was distinguishable.
Conclusions :
To conclude, our results suggest that cell phenotype is linked to the secretion of certain proteases/PIs in both groups. However, there seems to be differences in secretion of particular proteases and PIs between FECD and healthy specimens as morphology did not have a similar influence. These differences might initiate an imbalance between proteases and PIs explaining the irregular thickening of the Descemet membrane seen in FECD.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.