Establishment and Maintenance of the Pluripotent State

摘要 多能干细胞可以在体内产生所有类型的细胞，因此具有巨大的潜力 再生医学，并为发育生物学和药理学的研究提供了强有力的工具。 体细胞直接转化为多能(诱导多能干细胞：iPS)细胞的研究进展 这是产生大量定制干细胞的诱人途径。在以下方面取得了显著进展 高效地生成iPS单元，包括无集成线路，但是，详细的机械洞察 多能性的建立、随后的维持和有效的退出仍然缺乏。 在过去的四年里，我们的项目团队为实现更好的 对这些的理解(Smith等人自然，2012，14；Ziller等人。《自然》2013，14；Gifford等人。单元格2013； Tsankov等人。《自然》2015；廖添丁等人。自然遗传学2015年；Cacchiarelli等人在2015年及更多)。AS 不出所料，这些结果帮助确定了需要进一步试验的领域，而且 为我们提出了一系列新的挥之不去的问题，这些问题是我们理解人类多能性的核心。 我们在更大计划项目(过去和未来)框架内的目标将为以下方面提供重要的见解 这一结果将对人类重新编程和多能性领域产生广泛影响。 在接下来的几年里，我们将应用一种复杂的，最近工程的DNMT1基因敲除/可诱导的救援人类 ES细胞系将解决人类干细胞生物学中的基本问题，重点是天真与 启动状态。我们将应用单细胞技术和分析的最新进展，以获得前所未有的 对人类多能性生物学的洞察。最后，我们将使用我们最近开发和表征的 人类二次重新编程系统的定义和研究最关键的点之一(最终 在重新编程过程中)，这也是最不被理解的。

Abstract Pluripotent stem cells can give rise to all cell types in the body and have therefore enormous potential for regenerative medicine, and provide a powerful tool for studies in developmental biology and pharmacology. Advances in transforming somatic cells directly into pluripotent (induced pluripotent stem: iPS) cells provide an attractive avenue for generating large numbers of customized stem cells. Notable progress has been made to efficiently generate iPS cells, including integration-free lines, however, detailed mechanistic insights regarding the establishment, subsequent maintenance and efficient exit from pluripotency are still lacking. Over the past four years our program project team has made significant contributions towards a better understanding of these (Smith et al. Nature 2012,14; Ziller et al. Nature 2013,14; Gifford et al. Cell 2013; Tsankov et al. Nature 2015; Liao et al. Nature Genetics 2015; Cacchiarelli et al Cell in 2015 and more). As would be expected the results have helped pinpoint areas that need further experimentation and also presented us with a new set of lingering questions that are central to our understanding of human pluripotency. Our aims in the framework of the larger program project (past and future) will provide crucial insights towards this end and as a result will have a broad impact on the fields of human reprogramming and pluripotency. Over the next years, we will apply a complex, recently engineered DNMT1 knockout/inducible rescue human ES cell line to address fundamental questions in human stem cell biology with a focus on the naïve versus primed states. We will apply the latest advances in single cell technology and analysis to gain unprecedented insights into the biology of human pluripotency. Lastly, we will use our recently developed and characterized human secondary reprogramming system to define and study one of the most critical points (the final transition) in the reprogramming process that is also the least understood.