Project 2: Large ncRNAs and Epigenetic Regulation in Pluripotency

我们对干细胞生物学的理解的最新进展为医学提供了前所未有的希望。 进步。例如，现在可以将来自人类皮肤的成纤维细胞转化为诱导的成纤维细胞。 多能干细胞（iPSC）通过诱导四个主要的调节蛋白。反过来，这些因素调节着 复杂的舞蹈，重塑分化的表观遗传景观的多能状态。这 这一过程为无限量供应用于移植医学、药物和生物医学的基因定制细胞类型铺平了道路。 人类疾病的发现和研究。 虽然在理解IPSC所需的关键蛋白质编码因子方面取得了一些进展， 对指导这一过程的微调基因开关的了解要少得多， 保持多能状态。我们最近证明，大的基因间非编码RNA lincRNA（lincRNA）可以在维持关键细胞状态如多能性中充当这样的开关。我确 最近发现了人类iPSC重编程过程的lincRNA调控的新方面。 具体来说，我们鉴定了重编程所需的lincRNA-RoR（Regulator of Reprogramming）， 人成纤维细胞转化为iPSC。与这一观点进一步一致的是我们最近的三项额外发现， 报道：（i）超过100种lincRNA直接受“核心干细胞”转录因子（Oct 4，Sox 2 和Nanog）;（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，许多表达其特异性;细胞开关，（iii）lincRNA表现出独特的基因- 表达谱类似于少数已知的主多能性开关。总的来说，这些 研究表明，由重编程因子启动的功能重要的调节级联反应， 激活lincRNA，其具有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潜力， 成功地完成了重新编程过程。在这里，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节中的功能作用，以及（3）它们在重编程和表观遗传调节中的作用。 生化机制 相关性（参见说明）： 诱导多能干细胞是疾病建模、药物筛选和治疗的潜在革命性工具。 再生医学该建议旨在充分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，这是一个新的研究领域。 确保其在生物医学中的使用是有效和安全的重要步骤

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