Availability of mRNA Obtained from Peripheral Blood Mononuclear Cells for Testing Mutation Consequences in Dystrophic Epidermolysis Bullosa.

Dystrophic epidermolysis bullosa (DEB) is an inheritable blistering disease caused by mutations in , which encodes type VII collagen. To address the issue of genotype-phenotype correlations in DEB, analyzing the consequences of mutations using mRNA is indispensable. Herein we established a novel method for testing the effect of mutations in DEB using mRNA extracted from peripheral blood mononuclear cells (PBMCs). We investigated the consequences of four mutations (c.6573 + 1G > C, c.6216 + 5G > T, c.7270C > T and c.2527C > T) in three Japanese individuals with recessive DEB. The novel method detected the consequences of two recurrent mutations (c.6573 + 1G > C, c.6216 + 5G > T) and a novel mutation (c.7270C > T) accurately. In addition, it detected aberrant splicing resulting from a mutation (c.2527C > T) which was previously reported as a nonsense mutation. Furthermore, we revealed that type VII collagen-expressing cells in PBMCs have similar cell surface markers as mesenchymal stem cells; they were CD105, CD29, CD45, and CD34, suggesting that a small number of mesenchymal stem cells or mesenchymal stromal cells are circulating in the peripheral blood, which enables us to detect mRNA in PBMCs. Taken together, our novel method for analyzing mutation consequences using mRNA obtained from PBMCs in DEB will significantly contribute to genetic diagnoses and novel therapies for DEB.