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.
Highlights
To 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
Liovic et al [12] investigated the response of the EBS keratinocyte cell lines KEB4 and KEB7 (severe EBS Dowling-Meara (EBS-DM) phenotype, K14 mutation R125P) to hypo-osmotic stress in comparison to the wild-type cell line NEB1, and found dual-specificity phosphatases and their downstream targets ERK and p38 to be differentially regulated in EBS cells
We recently described the gene expression profile of KEB7 cells after applying suppression subtractive hybridization, and found dysregulated genes involved in keratinocyte differentiation, migration and wound healing [15]
Summary
To 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. KRT5 and KRT14 are the main stress-absorbing keratins in basal keratinocytes of the epidermis and related stratified epithelia These rod-shaped proteins form heterodimeric units that interact to build up the cytoskeletal intermediate filament (IF) network, a resilient yet adaptable scaffold that maintains cellular structural stability and, in turn, normal skin integrity and function. We follow-up that analysis with a more expansive expression profiling study, combining whole-transcriptome microarray examination with bioinformatics-assisted functional clustering, complementary qRT-PCR validation, and western blotting analysis of selected hits. Using this approach we were able to verify candidate genes previously described as being differentially expressed in EBS-DM, and to discover other differentially regulated genes not previously implicated in EBS pathology.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
