Assistant Professor No.1, University Road Tainan, Taiwan (Republic of China)
Abstract: Recessive Dystrophic Epidermolysis Bullosa (RDEB) is a severe genetic skin disorder caused by mutations in the COL7A1 gene, leading to truncated type VII collagen (C7) at the epidermal–dermal junction and resulting in extensive blistering, large non-healing wounds, and increased squamous cell carcinoma risk. In this study, patient-derived induced pluripotent stem cells (iPSCs) harboring the codon 1573 C→T (R525Ter) mutation, which introduces a premature stop codon at exon 12, were used alongside wild-type iPSCs (SC81103). Both were differentiated into fibroblasts (FB) and keratinocytes (KC), then combined with extracellular matrix and seeded onto polydimethylsiloxane (PDMS) devices to form a 3D microtissue-based skin organoid. Differentiation was confirmed by immunofluorescence staining of COL1A1/Vimentin (FB) and KRT5 (KC) and by real-time PCR of lineage markers (FB: COL1A1, COL3A1; KC: KRT5, KRT14, TP63). Further characterization using embryoid bodies (EBs) and single-cell assays supported the transition toward KC identity. These organoids recapitulated key molecular and structural features of healthy and RDEB skin, serving as a versatile in vitro model for disease mechanism studies and therapeutic testing. Future work will employ CRISPR/Cas9 and adenine base editing to correct the COL7A1 mutation and rescue functional C7 expression, thereby highlighting a potential path toward definitive treatments for RDEB.
