Mutation Reduction of Alopecia-Related Genes in hiPSC Using miR-520d-5p

Abstract

Background: iPSCs, with many mutations or epigenetic modifications, are in a unique aging state and have pluripotent properties. Previously, we reported that hsa-miR-520d-5p reverted fibroblasts to mesenchymal stem cells (MSCs) and converted approximately half of the mutations in cancer cells back to wild-type alleles. Objective: We focused on the genes involved in the hair life cycle comprising 4 phases and examined whether 520d-5p can convert iPSCs with mutation(s) to less-mutated status during the differentiation to MSCs. Materials and methods: Using next-generation sequencing analysis, we compared the effect of 520d-5p on the mutation level in 177 hair life cycle-related genes in iPSCs to that in iPSC-differentiated MSCs. Results: Transfection by hsa-miR-520d-5p induced genetic improvement in 79.7% (141/177) of the genes and reverted 17.7% (25/141) to wild type; the average conversion rate was 54.4%. The hsa-miR-520d-5p-induced genes, including the function of dermal papilla cells or bulge stem cells closer to intact status. Interestingly, the dramatic changes of nucleotide extended to non-target genes as well as target genes. Conclusions: Thus, hsa-miR-520d-5p is useful for studying hair or scalp conditioning and developing therapy against aging or hair diseases such as alopecia.

Keywords

• mutation
• iPSC
• MSC
• 520d-5p
• hair life cycle
• gene expression