Project 2: Large ncRNAs and Epigenetic Regulation in Pluripotency

我们对干细胞生物学理解的最新进展为医学带来了前所未有的希望 进步。例如，现在可以将人类皮肤的成纤维细胞转化为诱导的 通过诱导四种主调节蛋白来产生多能干细胞（iPSC）。反过来，这些因素调节 复杂的编排将差异化的表观遗传景观重塑为多能状态。这个 该过程为移植医学、药物的基因定制细胞类型的无限供应铺平了道路 人类疾病的发现和研究。 尽管在理解 IPSC 所需的关键蛋白质编码因子方面已经取得了一些进展 对于引导这一过程的精细调节的基因开关，人们对重新编程知之甚少。 维持多能状态。我们最近证明，大的基因间非编码RNA （lincRNA）可以充当维持关键细胞状态（例如多能性）的开关。确实，我们 最近发现了 lincRNA 调节人类 iPSC 重编程过程的一个新方面。 具体来说，我们确定了重编程所需的 lincRNA-RoR（重编程调节器） 人成纤维细胞转化为 iPSC。与这个想法进一步一致的是我们最近的另外三个发现 据报道：(i) 超过 100 种 lincRNA 直接受“核心干细胞”转录因子（Oct4、Sox2）调节 和纳诺格）； (ii) lincRNA 与维持分化的关键染色质修饰复合物相互作用 细胞状态，许多都表达了它们的特异性；细胞开关，(iii) lincRNA 表现出独特的基因- 表达谱与少数已知的主多能性开关相似。总的来说，这些 研究表明，由重编程因子引发的功能上重要的调控级联， 激活有可能与下游表观遗传机制相互作用并调节下游表观遗传机制的 lincRNA 成功完成重新编程过程。这里我们的目标是（1）全面鉴定lincRNA 参与重编程（2）它们在重编程和表观遗传调控中的功能作用以及（3）它们 生化机制。 相关性（参见说明）： 诱导多能干细胞是一种潜在的革命性工具，可用于疾病建模、药物筛选和 再生医学。该提案旨在全面表征大基因间非编码RNA的作用 （lincRNA）及其与建立多能性和表观遗传状态的功能相关性，这是一个 确保其在生物医学中的使用有效和安全的重要一步

Recent advances in our understanding of stem cell biology offer unprecedented hope for medical advancement. For example, it is now possible to transform fibroblasts from human skin to induced pluripotent stem cells (iPSCs) by induction of four master regulatory proteins. In turn these factors regulate a complex choreography that remodels the differentiated epigenetic landscapes to the pluripotent state. This process paves the way for limitless supply of genetically tailored cell types for transplantation medicine, drug discovery and the study of human disease. Although some progress has been made in understanding the key protein coding factors needed for IPSC reprogramming much less is known about the finely tuned genetic switches that guide this process and maintain the pluripotent state. We have recently demonstrated that large intergenic non-coding RNAs (lincRNAs) may serve as such switches in maintaining key cellular states such as pluripotency. Indeed, we recently discovered a new facet of lincRNA regulation of the human iPSC reprogramming process. Specifically, we identified lincRNA-RoR (Regulator of Reprogramming) that is required for reprogramming human fibroblast to iPSCs. Further consistent with this idea are three additional findings we recently reported: (i) Over 100 lincRNAs are directly regulated by the 'core stem-cell' transcription factors (Oct4, Sox2 and Nanog); (ii) lincRNAs interact with key chromatin modifying complexes that maintain the differentiation states of cells, and many convey their specificity; cell switches, (iii) lincRNAs exhibit distinctive gene- expression profiles similar to those of the few known master pluripotency switches. Collectively, these studies demonstrate a functionally important regulatory cascade initiated by reprogramming factors, which activate lincRNAs that have the potential to interface with and modulate downstream epigenetic machinery to successfully complete the reprogramming process. Here we aim to (1) Comprehensively identify lincRNAs involved in reprogramming (2) their functional roles in reprogramming and epigenetic regulation and (3) Their biochemical mechanisms. RELEVANCE (See instructions): Induced pluripotent stem cells is a potential revolutionary tool for disease modeling, drug screening and regenerative medicine. This proposal aims to fully characterize the roles of large intergenic non-coding RNAs (lincRNAs) and their functional relevance to establishing pluripotency and epigenetic states, which is an essential step towards ensuring that their use in biomedicine is effective and safe