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Single-Cell RNA Sequencing of TGF-Treated Human iPSC-Derived Skin Organoids

ABSTRACT: Human iPSC-derived skin organoids provide a powerful model for studying skin development and disease. Here, we investigated the effects of TGF-β treatment on human iPSC-derived skin organoids using single-cell RNA sequencing (scRNA-seq). Transcriptomic analysis identified diverse cellular populations, including keratinocytes and fibroblast-like cells, and revealed significant TGF-β-induced changes in cell states and gene expression. TGF-β treatment upregulated profibrotic and extracellular matrix-associated genes and activated pathways related to tissue remodeling and epithelial–mesenchymal transition. These findings demonstrate that iPSC-derived skin organoids recapitulate key features of TGF-β-mediated skin remodeling and provide a useful platform for studying fibrosis and cellular heterogeneity at single-cell resolution.

ORGANISM(S): Homo sapiens

PROVIDER: GSE330913 | GEO | 2026/06/10

REPOSITORIES: GEO

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Project description:Recessive dystrophic epidermolysis bullosa (RDEB) is a genodermatosis characterized by fragile skin forming blisters that heal invariably with scars. It is due to mutations in the COL7A1 gene encoding type VII collagen, the major component of anchoring fibrils connecting the cutaneous basement membrane to the dermis. Identical COL7A1 mutations often result in inter- and intra-familial disease variability, suggesting that additional modifiers contribute to RDEB course. Here, we studied a monozygotic twin pair with RDEB presenting markedly different phenotypic manifestations, while expressing similar amounts of collagen VII. Genome-wide expression analysis in twins' fibroblasts showed differential expression of genes associated with TGF-β pathway inhibition. In particular, decorin, a skin matrix component with anti-fibrotic properties, was found to be more expressed in the less affected twin. Accordingly, fibroblasts from the more affected sibling manifested a profibrotic and contractile phenotype characterized by enhanced α-smooth muscle actin and plasminogen activator inhibitor 1 expression, collagen I release and collagen lattice contraction. These cells also produced increased amounts of proinflammatory cytokines interleukin 6 and monocyte chemoattractant protein-1. Both TGF-β canonical (Smads) and non-canonical (MAPKs) pathways were basally more activated in the fibroblasts of the more affected twin. The profibrotic behaviour of these fibroblasts was suppressed by decorin delivery to cells. Our data show that the amount of type VII collagen is not the only determinant of RDEB clinical severity, and indicate an involvement of TGF-β pathways in modulating disease variability. Moreover, our findings identify decorin as a possible anti-fibrotic/inflammatory agent for RDEB therapeutic intervention. Primary fibroblast cultures from biopsies from two twins affected by recessive dystrophic epidermolysis bullosa were analyzed. Each hybridization was performed in biological triplicate and in technical duplicate.

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Single-Cell RNA Sequencing of TGF-Treated Human iPSC-Derived Skin Organoids

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