Signatures of Dermal Fibroblasts from RDEB Pediatric Patients.

Arkadii K Beilin,Daniil K Lukyanov,Kirill V Savostyanov,Eduard T Ambarchian,Nikolay N Murashkin,Nadezhda A Evtushenko,Alexander A Pushkov,Nadya G Gurskaya,Olga S Rogovaya,Andrey V Vasiliev,Ekaterina A Vorotelyak,Andrey P Fisenko

doi:10.3390/ijms22041792

Abstract

The recessive form of dystrophic epidermolysis bullosa (RDEB) is a debilitating disease caused by impairments in the junctions of the dermis and the basement membrane of the epidermis. Mutations in the COL7A1 gene induce multiple abnormalities, including chronic inflammation and profibrotic changes in the skin. However, the correlations between the specific mutations in COL7A1 and their phenotypic output remain largely unexplored. The mutations in the COL7A1 gene, described here, were found in the DEB register. Among them, two homozygous mutations and two cases of compound heterozygous mutations were identified. We created the panel of primary patient-specific RDEB fibroblast lines (FEB) and compared it with control fibroblasts from healthy donors (FHC). The set of morphological features and the contraction capacity of the cells distinguished FEB from FHC. We also report the relationships between the mutations and several phenotypic traits of the FEB. Based on the analysis of the available RNA-seq data of RDEB fibroblasts, we performed an RT-qPCR gene expression analysis of our cell lines, confirming the differential status of multiple genes while uncovering the new ones. We anticipate that our panels of cell lines will be useful not only for studying RDEB signatures but also for investigating the overall mechanisms involved in disease progression.

Highlights

We report a set of differentially expressed genes (DEG) between the recessive form of dystrophic epidermolysis bullosa (RDEB) and healthy fibroblasts and compare them with DEG obtained from the publicly available RNA-seq analysis of RDEB fibroblasts
To find the common and specific traits of DEG patterns in RDEB, we focused on the RNA-seq data of other fibroblasts induced by the c.6527insC mutation that was found to be recurrent in the Spanish RDEB population [22]
In order to compare the patterns of RDEB mutation effects on cellular homeostasis, we analyzed the available RDEB fibroblasts’ RNA-seq data with the c.6527insC mutation

Summary

Introduction

Up to twenty-four genetic subtypes of epidermolysis bullosa (EB) have currently been described for the four main types of the disease. Dystrophic epidermolysis bullosa (DEB) is one of the most severe forms of EB. The skin tissue of DEB patients displays anomalies of anchoring fibrils (AF), blistering and separation of sublamina densa as a result of mutations in the COL7A1 gene [1]. We focus on RDEB, which includes forms that vary in the severity of disease manifestations. There are less severe subtypes of RDEB with localized skin lesions or mucosal involvement. More than 800 different mutations have been identified in the COL7A1 gene, localizing in different parts of the sequence and leading to cutaneous manifestations of varying severity

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