Abstract
Purpose:
To elucidate gene expression patterns at different developmental stages of human corneal endothelial cells (HCECs).
Methods:
RNA-Seq was performed on three adult and two fetal HCEC tissues. To determine differential expression patterns between adult and fetal CECs from other cell types, and identify genes that were specifically overexpressed in either adult or fetal CECs, statistical analysis comparing the HCEC RNA-Seq reads to RNA-Seq reads from 12 different types of human cells was performed. Quantitative RT-PCR (Q-PCR) using RNA from freshly isolated HCECs and immunohistochemistry (IHC) analysis on frozen CE tissue sections was used to validate the RNA-Seq findings.
Results:
Eight TGF-beta signaling pathway members, and the non-canonical Wnt pathway genes Wnt5a and Wnt4 were upregulated in fetal corneal endothelium. The calcium ion binding proteins S100A4 and S100A6 were highly expressed in adult, but not fetal CECs. Additional genes that are differentially expressed in fetal and adult HCECs were also identified and confirmed at the mRNA and protein level.
Conclusions:
There is a shift in gene expression at different stages of HCEC development. Wnt signaling might play a rule during maturation of CEC.
Keywords: 481 cornea: endothelium •
533 gene/expression