Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 761001, Israel; New York Genome Center, New York, NY, USA.
Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 761001, Israel.
Cell Stem Cell. 2019 Feb 7;24(2):328-341.e9. doi: 10.1016/j.stem.2018.11.014. Epub 2018 Dec 13.
The epigenetic dynamics of induced pluripotent stem cell (iPSC) reprogramming in correctly reprogrammed cells at high resolution and throughout the entire process remain largely undefined. Here, we characterize conversion of mouse fibroblasts into iPSCs using Gatad2a-Mbd3/NuRD-depleted and highly efficient reprogramming systems. Unbiased high-resolution profiling of dynamic changes in levels of gene expression, chromatin engagement, DNA accessibility, and DNA methylation were obtained. We identified two distinct and synergistic transcriptional modules that dominate successful reprogramming, which are associated with cell identity and biosynthetic genes. The pluripotency module is governed by dynamic alterations in epigenetic modifications to promoters and binding by Oct4, Sox2, and Klf4, but not Myc. Early DNA demethylation at certain enhancers prospectively marks cells fated to reprogram. Myc activity drives expression of the essential biosynthetic module and is associated with optimized changes in tRNA codon usage. Our functional validations highlight interweaved epigenetic- and Myc-governed essential reconfigurations that rapidly commission and propel deterministic reprogramming toward naive pluripotency.
诱导多能干细胞(iPSC)重编程的表观遗传动力学在正确重编程的细胞中,在高分辨率和整个过程中仍然很大程度上未被定义。在这里,我们使用 Gatad2a-Mbd3/NuRD 耗尽和高效的重编程系统来表征将小鼠成纤维细胞转化为 iPSC。我们获得了基因表达、染色质结合、DNA 可及性和 DNA 甲基化水平的动态变化的无偏高分辨率分析。我们鉴定了两个不同且协同的转录模块,这些模块主导着成功的重编程,与细胞身份和生物合成基因有关。多能性模块由启动子的表观遗传修饰的动态变化以及 Oct4、Sox2 和 Klf4 的结合所控制,但不包括 Myc。某些增强子的早期 DNA 去甲基化前瞻性地标记了注定要重新编程的细胞。Myc 活性驱动必需的生物合成模块的表达,并与 tRNA 密码子使用的优化变化相关。我们的功能验证强调了相互交织的表观遗传和 Myc 控制的基本重构,这些重构迅速委托并推动确定性重编程向原始多能性发展。
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