308 The new cell cultures from dystrophic epidermolysis bullosa patients in Russia

Abstract

Dystrophic epidermolysis bullosa (DEB) is an inherited genetic disorder caused by mutations in COL7A1 gene. Dysfunction of type-VII collagen leads to extreme skin sensitivity to minor mechanical impact which results in the formation of sub lamina densa blisters. The data concerning the distribution of epidermolysis bullosa mutations in Russia are insufficient. This problem becomes especially important because of the wide geographic distribution of population and diverse national genetic composition in Russia. In this study, we describe cases of DEB in two children (7 and 16 years old) from different areas of the European part of Russia. Target regions of all currently known genes with mutations leading to EB symptom complex were investigated by the massive parallel sequencing. In the first patient, a missense variant in homozygous state was found in COL7A1 gene. The second patient was a compound heterozygous by the splice site variant and missense variant in COL7A1 gene. All three mutations have been previously described for the dystrophic type of EB with an autosomal recessive inheritance mechanism in HGMD professional database. The establishment of patient-specific cell lines with unlimited lifespan meets the needs of regenerative medicine. For this aim, we extract primary dermal fibroblasts from patients skin biopsies for reversible immortalization. cDNA of human telomerase reverse transcriptase (hTERT) was integrated into the genomes by lentivirus transduction. The plasmid provides the possibility for Cre-dependent control of hTERT insertion. Therefore, we obtained unique DEB cell cultures from Russian patients which are suitable for DEB modeling in vitro and testing new treatment approaches for DEB.