DETERMINANTS OF SELF-RENEWAL, DIFFERENTIATION, AND REPROGRAMMING OF HESCS

HECS 自我更新、分化和重新编程的决定因素

• 批准号: 8173148
• 负责人: James Alexander Thomson
• 金额: $ 4.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8173148
• 关键词: Cell Nucleus; Cells; Commit; Computer Retrieval of Information on Scientific Projects Database; Development; Epigenetic Process; Event; Funding; Genes; Grant; Histone Code; Histone H3; Individual; Institution; Left; Link; Maps; Mass Spectrum Analysis; Mediating; Memory; Myelogenous; Myeloid Cells; Outcome; Post-Translational Protein Processing; Process; Proliferating; Regenerative Medicine; Reporting; Research; Research Personnel; Resources; Role; Source; Specific qualifier value; Techniques; Time; Transact; Transplantation; United States National Institutes of Health; Variant; cell type; embryonic stem cell; histone modification; human embryonic stem cell; novel; nuclear reprogramming; pluripotency; promoter; self-renewal

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: Human ES cells are special because they can grow without limit and can give rise to all other cell types. Here we will try to understand why human ES cells have this remarkable developmental potential, and develop conditions to convert a cell with a more limited potential to an ES cell. Such reprogramming has implications for transplantation and regenerative medicine. Understanding how human embryonic stem (ES) cells can proliferate without limit and yet retain the ability to differentiate to any cell type is the theme that links the three individual projects of this proposal. Project 1 will identify novel histone modifications in human ES cells using a new mass spectrometry technique that allows an unprecedented ability to identify and map posttranslational protein modifications. Histone modifications in human ES cells will be identified globally, at promoters, and at select genes directly regulated by critical pluripotency factors. We will also examine the role of histone H3 variants in establishing long-term epigenetic memory. These studies will determine whether there is a novel histone code in pluripotent cells and determine how histone modifications change dynamically during differentiation. Project 2 examines the critical events that occur in the window of time during which human ES cells commit to exit the pluripotent state upon BMP-induced differentiation. This project will provide an increased understanding of the processes that commit human ES cells to exit the pluripotent state and specify different lineage outcomes. Project 3 will identify combinations of ES cell-specific genes that can reprogram differentiated cells to a pluripotent state. We have previously reported that when myeloid cells are fused with human ES cells, the myeloid nucleus is reprogrammed to an ES cell state, indicating that transacting factors in ES cells are sufficient to mediate nuclear reprogramming. Our preliminary results suggest that over expressing combinations of human ES cell-enriched genes can reprogram myeloid cells, and this project will optimize this reprogramming. The combination of these projects will provide an increased understanding of the pluripotent state and the basic processes by which a cell can leave or return to that state. Such an understanding will be important to transplantation and regenerative medicine.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 目的：人类胚胎干细胞很特殊，因为它们可以无限生长，并且可以产生所有其他细胞类型。在这里，我们将尝试了解为什么人类 ES 细胞具有如此显着的发育潜力，并开发条件将潜力更有限的细胞转化为 ES 细胞。这种重编程对移植和再生医学具有重要意义。 了解人类胚胎干 (ES) 细胞如何无限制地增殖，同时保留分化为任何细胞类型的能力，这是连接该提案的三个单独项目的主题。项目 1 将使用一种新的质谱技术来识别人类 ES 细胞中的新型组蛋白修饰，该技术具有前所未有的识别和绘制翻译后蛋白质修饰图谱的能力。人类 ES 细胞中的组蛋白修饰将在启动子处以及由关键多能性因子直接调节的选定基因处进行全面鉴定。我们还将研究组蛋白 H3 变体在建立长期表观遗传记忆中的作用。这些研究将确定多能细胞中是否存在新的组蛋白密码，并确定组蛋白修饰在分化过程中如何动态变化。项目 2 检查人类 ES 细胞在 BMP 诱导分化后退出多能状态的时间窗口内发生的关键事件。该项目将加深对人类 ES 细胞退出多能状态并指定不同谱系结果的过程的理解。项目 3 将鉴定 ES 细胞特异性基因的组合，这些基因可以将分化细胞重编程为多能状态。我们之前报道过，当骨髓细胞与人ES细胞融合时，骨髓核被重编程为ES细胞状态，表明ES细胞中的反式作用因子足以介导核重编程。我们的初步结果表明，人类 ES 细胞富集基因的过度表达组合可以重编程骨髓细胞，而该项目将优化这种重编程。这些项目的结合将加深对多能状态以及细胞离开或返回该状态的基本过程的理解。这种理解对于移植和再生医学非常重要。