Purchase this article with an account.
An-Katrien De Roo, Beatrijs Foets, Joost J van den Oord; Dendritic cells in Fuchs' endothelial corneal dystrophy. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5273.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Fuchs' endothelial corneal dystrophy (FECD) is the major indication for corneal endothelial transplantation. However, there is a worldwide shortage of donor corneas, and the prevalence of FECD is expected to rise with aging of the population. To develop alternative therapies, we need a better understanding of the molecular pathogenesis of FECD.
We compared gene expression profiles in human corneal endothelium (HCEn) of patients with FECD (n = 4) and normal donors (n = 4), using microarray expression analysis (MEA) and Ingenuity Pathway Analysis (IPA). The expression of 179 genes of interest was validated with reverse transcriptase quantitative PCR (RT-qPCR) on HCEn of patients with FECD (n = 9) and normal donors (n = 8). RT-qPCR data were analyzed with the ΔΔCT method, using RPL13A, RPL19 and RPS5 as reference genes. Further validation consisted of immunohistochemistry (IHC) and immunofluorescence (IF) on HCEn whole mounts on the one hand, and of IHC on paraffin sections of full-thickness corneas from patients with FECD (n = 6) and controls (n = 6, enucleations for uveal melanoma) on the other hand.
Unsupervised analysis of the MEA data could easily separate patients with FECD from normal donors. IPA identified pathways, functions and networks related to dendritic cell maturation, antigen presentation and HLA-DR. There was a 65% concordance between RNA-expression on micro-array and on RT-qPCR, using |log2(FECD/normal)| ≥ 1 and p < 0.05 as cut-off values for significant differential expression. IHC and IF whole mount stainings revealed the presence of dendritic cells in all examined cases of FECD but not in normal donor HCEn (n = 13). These dendritic cells clustered around guttae, embracing them with their long processes. Furthermore, they expressed CD45, indicating bone-marrow origin, as well as HLA-DRA and alpha smooth muscle actin (αSMA). This phenotype is suggestive of circulating fibrocytes, which are involved in various interstitial chronic fibrosing disorders.
We have identified the presence of dendritic cells in corneal endothelium from patients with FECD. These dendritic cells co-express HLA-DRA, CD45 and αSMA. To our knowledge these cells have not previously been described in FECD. We are now further investigating their origin and their role in the pathogenesis of FECD.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
This PDF is available to Subscribers Only