Incomplete MyoD-induced transdifferentiation is associated with chromatin remodeling deficiencies

• Published in: Nucleic acids research Vol. 45; no. 20; pp. 11684 - 11699
• Main Authors: Manandhar, Dinesh, Song, Lingyun, Kabadi, Ami, Kwon, Jennifer B, Edsall, Lee E, Ehrlich, Melanie, Tsumagari, Koji, Gersbach, Charles A, Crawford, Gregory E, Gordân, Raluca
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 16.11.2017
• Subjects: Blotting, Western; Cell Transdifferentiation - genetics; Cells, Cultured; Cellular Reprogramming - genetics; Chromatin - genetics; Chromatin - metabolism; Chromatin Assembly and Disassembly - genetics; Fibroblasts - cytology; Fibroblasts - metabolism; Gene Expression Profiling - methods; Gene Ontology; Gene regulation, Chromatin and Epigenetics; HEK293 Cells; Humans; Microscopy, Confocal; MyoD Protein - genetics; MyoD Protein - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Skin - cytology
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/gkx773

Our current understanding of cellular transdifferentiation systems is limited. It is oftentimes unknown, at a genome-wide scale, how much transdifferentiated cells differ quantitatively from both the starting cells and the target cells. Focusing on transdifferentiation of primary human skin fibroblasts by forced expression of myogenic transcription factor MyoD, we performed quantitative analyses of gene expression and chromatin accessibility profiles of transdifferentiated cells compared to fibroblasts and myoblasts. In this system, we find that while many of the early muscle marker genes are reprogrammed, global gene expression and accessibility changes are still incomplete when compared to myoblasts. In addition, we find evidence of epigenetic memory in the transdifferentiated cells, with reminiscent features of fibroblasts being visible both in chromatin accessibility and gene expression. Quantitative analyses revealed a continuum of changes in chromatin accessibility induced by MyoD, and a strong correlation between chromatin-remodeling deficiencies and incomplete gene expression reprogramming. Classification analyses identified genetic and epigenetic features that distinguish reprogrammed from non-reprogrammed sites, and suggested ways to potentially improve transdifferentiation efficiency. Our approach for combining gene expression, DNA accessibility, and protein-DNA binding data to quantify and characterize the efficiency of cellular transdifferentiation on a genome-wide scale can be applied to any transdifferentiation system.