Chromatin modifying enzymes as modulators of reprogramming
By T. Onder, Nergis Kara, Anne B. C. Cherry, Amit U. Sinha, Nan Zhu, K. Bernt, Patrick Cahan, O. Mancarci, J. Unternaehrer, Piyush B. Gupta and 3 more. Nature, 2012
675 citations recorded by the source corpus.
Generation of induced pluripotent stem cells (iPSCs) by somatic cell reprogramming involves global epigenetic remodelling. Whereas several proteins are known to regulate chromatin marks associated with the distinct epigenetic states of cells before and after reprogramming, the role of specific chromatin-modifying enzymes in reprogramming remains to be determined. To address how chromatin-modifying proteins influence reprogramming, we used short hairpin RNAs (shRNAs) to target genes in DNA and histone methylation pathways, and identified positive and negative modulators of iPSC generation. Whereas inhibition of the core components of the polycomb repressive complex 1 and 2, including the histone 3 lysine 27 methyltransferase EZH2, reduced reprogramming efficiency, suppression of SUV39H1, YY1 and DOT1L enhanced reprogramming. Specifically, inhibition of the H3K79 histone methyltransferase DOT1L by shRNA or a small molecule accelerated reprogramming, significantly increased the yield of iPSC colonies, and substituted for KLF4 and c-Myc (also known as MYC). Inhibition of DOT1L early in the reprogramming process is associated with a marked increase in two alternative factors, NANOG and LIN28, which play essential functional roles in the enhancement of reprogramming. Genome-wide analysis of H3K79me2 distribution revealed that fibroblast-specific genes associated with the epithelial to mesenchymal transition lose H3K79me2 in the initial phases of reprogramming. DOT1L inhibition facilitates the loss of this mark from genes that are fated to be repressed in the pluripotent state. These findings implicate specific chromatin-modifying enzymes as barriers to or facilitators of reprogramming, and demonstrate how modulation of chromatin-modifying enzymes can be exploited to more efficiently generate iPSCs with fewer exogenous transcription factors.
Subfields: Molecular Biology
View Paper: https://www.semanticscholar.org/paper/fa78799805ee209aac5ede2b7655fe5c770a3f7f
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Citations in the corpus
- Facilitators and Impediments of the Pluripotency Reprogramming Factors’ Initial Engagement with the Genome (2012, positive): “A recent screen of 21 histone modifiers showed that knocking down SUV39H1, which elicits H3K9me3 in heterochromatin, enhances reprogramming markedly (Onder et al., 2012 ), but the basis for the effect was not clear.”
- Occupying chromatin: Polycomb mechanisms for getting to genomic targets, stopping transcriptional traffic, and staying put (2013, background): “…also great interest in the roles of PcG proteins in the transcriptional circuitry of embryonic and adult stem cells (Boyer et al., 2006; Ezhkova et al., 2009), which is underscored by recent advances that implicate PcG functions in cellular reprogramming (Onder et al., 2012; Pereira et al., 2010).”