High throughput gene expression analysis identifies reliable expression markers of human corneal endothelial cells

PLoS One. 2013 Jul 2;8(7):e67546. doi: 10.1371/journal.pone.0067546. Print 2013.

Abstract

Considerable interest has been generated for the development of suitable corneal endothelial graft alternatives through cell-tissue engineering, which can potentially alleviate the shortage of corneal transplant material. The advent of less invasive suture-less key-hole surgery options such as Descemet's Stripping Endothelial Keratoplasty (DSEK) and Descemet's Membrane Endothelial Keratoplasty (DMEK), which involve transplantation of solely the endothelial layer instead of full thickness cornea, provide further impetus for the development of alternative endothelial grafts for clinical applications. A major challenge for this endeavor is the lack of specific markers for this cell type. To identify genes that reliably mark corneal endothelial cells (CECs) in vivo and in vitro, we performed RNA-sequencing on freshly isolated human CECs (from both young and old donors), CEC cultures, and corneal stroma. Gene expression of these corneal cell types was also compared to that of other human tissue types. Based on high throughput comparative gene expression analysis, we identified a panel of markers that are: i) highly expressed in CECs from both young donors and old donors; ii) expressed in CECs in vivo and in vitro; and iii) not expressed in corneal stroma keratocytes and the activated corneal stroma fibroblasts. These were SLC4A11, COL8A2 and CYYR1. The use of this panel of genes in combination reliably ascertains the identity of the CEC cell type.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Anion Transport Proteins / genetics*
  • Anion Transport Proteins / metabolism
  • Antiporters / genetics*
  • Antiporters / metabolism
  • Autopsy
  • Biomarkers / metabolism
  • Collagen Type VIII / genetics*
  • Collagen Type VIII / metabolism
  • Corneal Keratocytes / cytology
  • Corneal Keratocytes / metabolism
  • Corneal Stroma / cytology
  • Corneal Stroma / metabolism
  • Endothelial Cells / cytology
  • Endothelial Cells / metabolism*
  • Endothelium, Corneal / cytology
  • Endothelium, Corneal / metabolism*
  • Female
  • Gene Expression Profiling
  • Gene Expression*
  • Humans
  • Male
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Middle Aged
  • Organ Specificity
  • Primary Cell Culture

Substances

  • Anion Transport Proteins
  • Antiporters
  • Biomarkers
  • COL8A2 protein, human
  • CYYR1 protein, human
  • Collagen Type VIII
  • Membrane Proteins
  • SLC4A11 protein, human

Grant support

This study is supported by Agency for Science, Technology and Research TCRP Grant (TCR0101673). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.