Molecular Analysis of ES Cells

ES 细胞的分子分析

• 批准号: 6927893
• 负责人: ALI H BRIVANLOU
• 金额: $ 33万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6927893
• 关键词: Xenopus; Xenopus oocyte; animal genetic material tag; animal tissue; biological signal transduction; biomarker; biotechnology; cell differentiation; cell growth regulation; cell population study; cell proliferation; developmental genetics; embryo /fetus; embryogenesis; embryonic stem cell; functional /structural genomics; gene expression; gene expression profiling; genetic regulation; human fetus tissue; human genetic material tag; laboratory mouse; microarray technology; molecular genetics; transforming growth factors

DESCRIPTION (provided by applicant): Embryonic stem cells are pluripotent cells derived from the epiblast of the early embryo. In culture they exhibit two important properties: they can maintain an undifferentiated state indefinitely, and they can be pushed to differentiate into a variety of cell types. This proposal targets the molecular dissection of pluripotency and self-renewal in both mouse and human embryonic stem cells (MESCs and HESCs), comparatively. We have recently obtained, and successfully cultured, five of the NIH-approved HESC lines (NIH codes WA01, BG01, BG02, ES03, and ES04). Our preliminary results provide a signature for the state of stemness in both ESCs. We have also demonstrated that the activation of the Wnt pathway is sufficient to maintain the undifferentiated phenotypes in both species. I propose two specific aims: 1- Refinement of the global and comparative molecular profiling of HESCs, and its comparison with MESCs. This allows the establishment of: (i) quantitative molecular standards, for undifferentiated HESCs, (ii) characterization of causal genes for the state of stemness, and (iii) measurement of molecular changes during differentiation paradigms. 2- Characterization of the role of signaling pathways underlying pluripotency, by (i) testing the activity of a novel small compound GSK3¿ inhibitor, BIO, in vivo in mouse embryos; (ii) addressing the role of TGF¿ signaling in this process; and (iii) performing an unbiased expression cloning to identify molecular factors involved in self-renewal in HESCs. In addition to providing a window into early human embryogenesis, HESCs could constitute a renewable source of a large variety of differentiated cells and be employed to replace diseased or damaged tissue by cellular transplantation. Therefore, the establishment of molecular standards and global molecular knowledge of their pluripotency is an obligatory step toward the design of rational clinical treatments.

描述（申请人提供）：胚胎干细胞是源自早期胚胎外胚层的多能细胞，在培养中它们表现出两个重要特性：它们可以无限期地维持未分化状态，并且它们可以被推动分化成多种细胞。相对而言，我们最近已经成功地获得了小鼠和人类胚胎干细胞（MESC 和 HESC）的多能性和自我更新的分子解剖。我们的初步结果为两种 ESC 的干性状态提供了标志。途径足以维持两个物种的未分化表型，我提出两个具体目标： 1- 完善 HESC 的整体和比较分子谱，及其与它的比较。这允许建立：(i) 未分化 HESC 的定量分子标准，(ii) 干性状态的因果基因的表征，以及 (iii) 分化范例期间分子变化的测量 2- 角色表征。通过 (i) 测试新型小化合物 GSK3 的活性，研究多能性背后的信号通路抑制剂，BIO，在小鼠胚胎体内的作用；（ii）解决 TGF¿ (iii) 进行无偏表达克隆以鉴定参与 HESC 自我更新的分子因子。因此，建立其多能性的分子标准和全球分子知识是设计合理临床治疗的必要步骤。