Chromatin-modifying enzymes as modulators of reprogramming.
Chromatin-modifying enzymes as modulators of reprogramming.
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DOI：
10.1038/nature10953
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发表时间：
2012-03-04
期刊：
影响因子：
64.8
通讯作者：
Daley, George Q.
中科院分区：
文献类型：
--
作者：
Onder, Tamer T.;Kara, Nergis;Cherry, Anne;Sinha, Amit U.;Zhu, Nan;Bernt, Kathrin M.;Cahan, Patrick;Mancarci, B. Ogan;Unternaehrer, Juli;Gupta, Piyush B.;Lander, Eric S.;Armstrong, Scott A.;Daley, George Q.
通过体细胞重编程产生诱导多能干细胞（iPSC）涉及整体表观遗传重塑。虽然已知几种蛋白质调节与重编程前后细胞的不同表观遗传状态相关的染色质标记，但特定染色质修饰酶在重编程中的作用仍有待确定。为了解决染色质修饰蛋白如何影响重编程，我们使用shRNA靶向DNA和组蛋白甲基化途径中的基因，并鉴定了iPSC生成的正和负调节剂。虽然抑制多梳抑制复合物1和2的核心组分，包括组蛋白3赖氨酸27甲基转移酶Ezh 2，降低了重编程效率，但抑制SUV 39 H1，YY 1和Dot 1 L增强了重编程。具体而言，通过shRNA或小分子抑制H3 K79组蛋白甲基转移酶Dot 1 L加速了重编程，显著增加了iPSC集落的产量，并取代了Klf 4和c-Myc。在重编程过程的早期抑制Dot 1 L与两个替代因子Nanog和Lin 28的显著增加有关，这两个因子在增强重编程中起着重要的功能作用。H3 K79 me 2分布的全基因组分析显示，与上皮向间质转化相关的成纤维细胞特异性基因在重编程的初始阶段丢失H3 K79 me 2。Dot 1 L抑制促进了该标记从注定在多能状态下被抑制的基因中的丢失。这些发现暗示特定的染色质修饰酶作为重编程的障碍或促进剂，并展示了如何利用染色质修饰酶的调节来更有效地产生具有更少外源转录因子的iPSC。
Generation of induced pluripotent stem cells (iPSCs) by somatic cell reprogramming involves global epigenetic remodeling. While 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 shRNAs to target genes in DNA and histone methylation pathways, and have identified positive and negative modulators of iPSC generation. While 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. 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.
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影响因子：
3.9
作者：
Stulemeijer IJ;Pike BL;Faber AW;Verzijlbergen KF;van Welsem T;Frederiks F;Lenstra TL;Holstege FC;Gasser SM;van Leeuwen F
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