Expression Analysis of Function-Related Proteins of Cultured Corneal Endothelial Cells Isolated from Patients with Fuchs Endothelial Corneal Dystrop

Abstract

Purpose: Fuchs endothelial corneal dystrophy (FECD) is a corneal pathology that affects the endothelial cell’s ability to maintain deturgescence, resulting in a progressive loss of corneal transparency. We have previously shown that cultured FECD cells are able to maintain corneal transparency when transplanted in a living feline eye (Haydari et al, IOVS 2012). The goal of this study was to characterize the expression of the function related proteins of a corneal endothelium tissue-engineered (TE) using cultured FECD cells (TE-FECD).

Methods: Corneal endothelial cells were isolated from surgical specimens (FECD; n=3) and age-matched normal Eye bank corneas (normal ; n=3), and cultured for 24 days to obtain a post-confluent monolayer of endothelial cells. Gene profiling experiments were performed on the mRNA of these cells using Agilent SurePrint G3 Human Gene expression microarrays. Cultured FECD (n=5) and normal (n=3) cells were seeded on devitalized corneas and cultured to TE corneas. Protein expression was assessed using immunofluorescence. Normal corneas (n=6) and FECD surgical specimens (n=4) were also used as native controls.

Results: The expression of many genes transcripts was deregulated in FECD compared to normal cultured cells (ATP1B1: +1.7X, AQP1: +3.9X, SLC4A4: +1.4X, SLC16A3: +1.5X). Protein expression was lower in native FECD specimens compared to native normal specimens for ATP1A1, AQP1, SLC4A4, SLC16A1 and SLC16A3. The protein expression of ATP1A1, ATP1B1, AQP1, SLC4A4 and SLC16A3 was also lower in all TE corneas (FECD and normal) compared to the normal native corneas. The expression profile of function-related proteins was similar in TE-FECD and TE-Normal corneas.

Conclusions: TE-FECD and TE- normal endothelia showed similar function-related protein expression in vitro, contrary to the difference observed between the normal and FECD native controls. These results support our experimental findings indicating that FECD cells rehabilitated in culture can recover the capacity of maintaining corneal transparency once transplanted in the living eye.

Keywords: 481 cornea: endothelium • 494 degenerations/dystrophies • 666 pump/barrier function