Whole-transcriptome gene expression profiling in an epidermolysis bullosa simplex Dowling-Meara model keratinocyte cell line uncovered novel, potential therapeutic targets and affected pathways

Abstract

BackgroundTo be able to develop effective therapeutics for epidermolysis bullosa simplex (EBS), it is necessary to elucidate the molecular pathomechanisms that give rise to the disease’s characteristic severe skin-blistering phenotype.ResultsStarting with a whole-transcriptome microarray analysis of an EBS Dowling-Meara model cell line (KEB7), we identified 207 genes showing differential expression relative to control keratinocytes. A complementary qRT-PCR study of 156 candidates confirmed 76.58 % of the selected genes to be significantly up-regulated or down-regulated (p-value <0.05) within biological replicates. Our hit list contains previously identified genes involved in epithelial cell proliferation, cell-substrate adhesion, and responses to diverse biological stimuli. In addition, we identified novel candidate genes and potential affected pathways not previously considered as relevant to EBS pathology.ConclusionsOur results broaden our understanding of the molecular processes dysregulated in EBS.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-015-1783-7) contains supplementary material, which is available to authorized users.

Prevalence, pathophysiology and management of itch in epidermolysis bullosa*

“…In six patients with EBS, gene expression analysis of the epidermis in normal‐appearing skin compared with an equal number of controls identified 28 differentially expressed genes; 68 the majority of these were implicated in fatty acid metabolism or the structural organization of the epidermis, which are both pertinent to the synthesis and integrity of the cornified cell envelope. Analogous differences in gene expression were identified in vitro between EBS cell lines and wildtype control keratinocytes 69,70 . Barrier dysfunction resulting from such modifications in genetic expression could make skin vulnerable to environmental pruritogens and lower the threshold for itch.…”

“…Analogous differences in gene expression were identified in vitro between EBS cell lines and wildtype control keratinocytes. 69,70 Barrier dysfunction resulting from such modifications in genetic expression could make skin vulnerable to environmental pruritogens and lower the threshold for itch. However, direct evidence for this hypothesis is still lacking for EB-associated itch.…”

Keratins as an Inflammation Trigger Point in Epidermolysis Bullosa Simplex

“…Other work with whole-transcriptome gene expression analysis, which was performed in the same model cell line (KEB7) with KRT14 R125P, uncovered some new specific targets affecting the EBS profile of gene expression, such as the significant downregulation of K19 [14]. Among other aspects, the positive regulation of BMP signaling and negative regulation of the canonical WNT receptor signaling pathways were shown.…”

“…The unfolded protein response (UPR) in a cell is activated by stress in the endoplasmic reticulum (ER) and initiates stress-induced homeostasis in the epidermis with chronic sterile inflammation, as in cases of other skin inflammatory diseases [12]. Several anomalies have been identified in gene cascades involved in cell proliferation, which are elements of the bone morphogenetic protein (BMP) signaling pathway, fatty acid metabolism, and retinoic acid signaling, as well as genes involved in the regulation of keratinization [13,14]. Additionally, microbial colonization aggravates the course of EBS.…”

Microbiopsy‐based minimally invasive skin sampling for molecular analysis is acceptable to Epidermolysis Bullosa Simplex patients where conventional diagnostic biopsy was refused

“…In this study, KRT5, KRT14 and GAPDH serve as a widely studied, proof of concept gene set to evaluate whether microbiopsy sampling in fragile skin can be used for molecular analysis. 11 Five clinically diagnosed EBS patients (n = 5) and 4 healthy volunteers (n = 4) were recruited. Table 1 shows that 4 out of 5 patients have refused to take a conventional skin biopsy procedure previously.…”