Abstract
Cell lineage conversion of fibroblasts to specialized cell types through transdifferentiation may provide a fast and alternative cell source for regenerative medicine. Here we show that transient transduction of fibroblasts with the four reprogramming factors (Oct4, Sox2, Klf4, and c-Myc) in addition to the early lung transcription factor Nkx2-1 (also known as Ttf1), followed by directed differentiation of the cells, can convert mouse embryonic and human adult dermal fibroblasts into induced lung-like epithelial cells (iLEC). These iLEC differentiate into multiple lung cell types in air liquid interface cultures, repopulate decellularized rat lung scaffolds, and form lung epithelia composed of Ciliated, Goblet, Basal, and Club cells after transplantation into immune-compromised mice. As proof-of-concept, differentiated human iLEC harboring the Cystic Fibrosis mutation dF508 demonstrated pharmacological rescue of CFTR function using the combination of lumacaftor and ivacaftor. Overall, this is a promising alternative approach for generation of patient-specific lung-like progenitors to study lung function, disease and future regeneration strategies.
Highlights
Cell lineage conversion of fibroblasts to specialized cell types through transdifferentiation may provide a fast and alternative cell source for regenerative medicine
While morphological changes were observed as early as 5 days after initiation of definitive endoderm (DE) differentiation, epithelial-like cells only emerged at the anterior ventral foregut equivalent stage
These groups of cells expanded into colonies (3–10 epithelial colonies per 104 cells representing a range of 0.03–0.1% conversion efficiency)
Summary
Cell lineage conversion of fibroblasts to specialized cell types through transdifferentiation may provide a fast and alternative cell source for regenerative medicine. We show that transient transduction of fibroblasts with the four reprogramming factors (Oct[4], Sox[2], Klf[4], and c-Myc) in addition to the early lung transcription factor Nkx[] ( known as Ttf1), followed by directed differentiation of the cells, can convert mouse embryonic and human adult dermal fibroblasts into induced lung-like epithelial cells (iLEC). Circumvent the iPS stage, studies have shown that overexpression of lineage-specific transcription factors can lead to direct lineage conversion of fibroblasts to neurons[9,10], cardiomyocytes[11], intestinal progenitors[12], renal tubular cells[13] and hepatocytes[14]. Many of these direct conversion studies have largely yielded immature, embryonic-like or cell phenotypes of an uncertain relationship to their in vivo counterparts
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