Project 2: Large ncRNAs and Epigenetic Regulation in Pluripotency

项目 2：大 ncRNA 和多能性表观遗传调控

• 批准号: 8379981
• 负责人: John Louis Rinn
• 金额: $ 47.05万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8379981
• 关键词: Anatomic Sites; Biochemical; Biological Assay; Cell physiology; Cells; Chromatin; Code; Complex; Computer Analysis; DNA; DNA Microarray Chip; Data Set; Disease model; Ensure; Epigenetic Process; Exhibits; Fibroblasts; Functional RNA; Gene Expression; Gene Expression Profile; Genetic; Genomics; Global Change; Histones; Human; Instruction; Maps; Measures; Mediating; Medical; Methods; Molecular Profiling; Monitor; Preclinical Drug Evaluation; Principal Investigator; Process; Proteins; RNA; RNA Sequences; Regenerative Medicine; Regulation; Reporting; Role; Site; Skin; Specificity; Stem cells; base; cell type; chromatin immunoprecipitation; cost effective; design; drug discovery; gain of function; genetic regulatory protein; histone modification; human disease; induced pluripotent stem cell; insight; pluripotency; research study; response; stem cell biology; tool; transcription factor; transplantation medicine

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

我们对干细胞生物学理解的最新进展为医学提供了前所未有的希望 进步。例如，现在可以将人皮肤的成纤维细胞转化为诱导性的 四种主要调节蛋白诱导的多能干细胞(IPSCs)。反过来，这些因素又调节着一个 将差异化的表观遗传景观重塑为多潜能状态的复杂编排。这 这一过程为无限供应用于移植药物、药物的基因定制细胞类型铺平了道路 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取得了一些进展 重新编程对指导这一过程的微调遗传开关知之甚少 保持全能状态。我们最近证明了大的基因间非编码RNA (LincRNAs)可以作为这样的开关来维持关键的细胞状态，如多能性。事实上，我们 最近发现了lincRNA调节人类IPSC重编程过程的一个新方面。 具体地说，我们确定了重新编程所需的lincRNA-ROR(重新编程的调节因子) 人成纤维细胞转化为ipscs。与这一想法进一步一致的是我们最近的三个额外发现 报道：(I)超过100个lincRNAs由核心干细胞转录因子(Oct4，Sox2)直接调控 和Nanog)；(Ii)lincRNAs与维持分化的关键染色质修饰复合体相互作用 细胞的状态，许多传递其特异性；细胞开关，(Iii)lincRNA显示独特的基因- 表达谱类似于少数已知的主多能开关的表达谱。总而言之，这些 研究表明，由重新编程因子启动的功能重要的调节级联反应， 激活有可能与下游表观遗传机制对接并调节其下游表观遗传机制的lincRNA 成功地完成了重新编程过程。在这里，我们的目标是(1)全面识别lincRNAs 参与重新编程(2)它们在重新编程和表观遗传调节中的功能作用以及(3)它们 生化机制。 相关性(请参阅说明)： 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。这项提议旨在充分表征大的基因间非编码RNA的作用 (LincRNAs)及其与建立多能性和表观遗传状态的功能相关性，这是一种 确保它们在生物医学中的使用是有效和安全的关键一步