One-step Reprogramming of Human Fibroblasts into Oligodendrocyte-like Cells by SOX10, OLIG2, and NKX6.2

• DOI: 10.1016/j.stemcr.2021.03.001
• Publication Date: 2021-03-25T14:36:53Z
• Authors: Konstantina Chano...; Benjamín Hernánde...; Farina Windener; Christian Thomas; Martin Stehling; Sabah Mozafari; Stefanie Albrecht; Linda Ottoboni; Jack Antel; Kee-Pyo Kim; Sergiy Velychko; Qiao Ling Cui; Yu Kang T. Xu; Gianvito Martino; Jürgen Winkler; Hans R. Schöler; Anne Baron-Van Ev...; Odile Boespflug-T...; Juan M. Vaquerizas; Marc Ehrlich; Tanja Kuhlmann
• Keywords: Pelizaeus-Merzbacher Disease; Transcription, Genetic; [SDV]Life Sciences [q-bio]; oligodendrocytes; Article; Cell Line; Epigenesis, Genetic; 03 medical and health sciences; Cell Movement; Humans; direct conversion; PMD; Gene Silencing; Transgenes; ATAC-seq; PMD; compound screenin; direct conversion; epigenetic age; human fibroblasts; oligodendrocytes; Myelin Sheath; Homeodomain Proteins; 0303 health sciences; epigenetic age; human fibroblasts; SOXE Transcription Factors; Age Factors; ATAC-seq; Fibroblasts; Oligodendrocyte Transcription Factor 2; Cellular Reprogramming; Chromatin Assembly and Disassembly; Chromatin; 3. Good health; Oligodendroglia; compound screenin

Limited access to human oligodendrocytes impairs better understanding of oligodendrocyte pathology in myelin diseases. Here, we describe a method to robustly convert human fibroblasts directly into oligodendrocyte-like cells (dc-hiOLs), which allows evaluation of remyelination-promoting compounds and disease modeling. Ectopic expression of SOX10, OLIG2, and NKX6.2 in human fibroblasts results in rapid generation of O4+ cells, which further differentiate into MBP+ mature oligodendrocyte-like cells within 16 days. dc-hiOLs undergo chromatin remodeling to express oligodendrocyte markers, ensheath axons, and nanofibers in vitro, respond to promyelination compound treatment, and recapitulate in vitro oligodendroglial pathologies associated with Pelizaeus-Merzbacher leukodystrophy related to PLP1 mutations. Furthermore, DNA methylome analysis provides evidence that the CpG methylation pattern significantly differs between dc-hiOLs derived from fibroblasts of young and old donors, indicating the maintenance of the source cells' "age." In summary, dc-hiOLs represent a reproducible technology that could contribute to personalized medicine in the field of myelin diseases.