Hormonal Control of Histone Modifications in ES Cells

ES 细胞中组蛋白修饰的激素控制

• 批准号: 7176168
• 负责人: MICHAEL K FRITSCH
• 金额: $ 27.07万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7176168
• 关键词: Acetylation; Acetylesterase; Aplastic Anemia; Biochemical; Biological Assay; Cell Differentiation process; Cell Extracts; Cells; Chimeric Proteins; Chromatin; Commit; CpG Islands; DNA; DNA Microarray Chip; DNA Microarray format; Data; Deacetylase; Development; Disease; Ectoderm; Genes; Global Change; Goals; Growth Factor; Hematopoietic; Histone Acetylation; Histone Deacetylase; Histone Deacetylase Inhibitor; Histone H3; Histones; Hormonal; Immunofluorescence Immunologic; In Vitro; LIF gene; Maps; Methods; Methylation; Modeling; Modification; Molecular; Parkinson Disease; Patients; Pattern; Population; Progesterone Receptors; Promoter Regions; Protein Overexpression; Proteins; Rate; Recording of previous events; Research Design; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Screening procedure; Signal Transduction; Standards of Weights and Measures; Teratogens; Testing; Time; Transplantation; Trichostatin A; Undifferentiated; Valproic Acid; Western Blotting; Withdrawal; day; design; embryonic stem cell; histone acetyltransferase; human RARB protein; improved; inhibitor/antagonist; leukemia inhibitory factor; novel; programs; promoter; relating to nervous system; stem cell technology

The molecular aspects of the earliest steps in embryonic stem (ES) cell differentiation remain poorly understood. Our preliminary data suggest that global histone acetylation may be a critical first step in differentiation. The goals of this proposal are to establish whether global histone acetylation and/or methytation occurs during multipie hormonally induced methods of ES cell differentiation and establish the time frame in which these histone modifications occur using standard assays for histone modifications. Our model predicts that the bulk of these histone modifications probably occur in promoter regions and we will make use of novel CpG island arrays to confirm this. Studies are designed to determine whether the global histone modifications that occur during exit from the undifferentiated ES cell state are uniquely different from the gene-specific histone modifications induced by hormonal signallingto highly differentiated cells. The second specific aim is to explore the possible mechanisms leading to these unique global histone modifications early in ES cell differentiation by screening ES cell extracts for various enzymatic activities responsible for covalent modification of histories. The third specific aim is designed to test the functional significance of histone modifications in directly regulating ES cell differentiation. The histone deacetylase inhibitor, trichostatin A (TSA), will be used in conjunction with specific growth factors to increase the rate and proportion of cells directed to a specific committed cell fate. In addition, overexpression of specific gene products designed to inhibit histone acetyltransferase activity or increase histone deacetylase activity in ES cells will be assessed for effects on the rate of ES cell differentiation and overall cell fate commitment. The proposed studies are designed to understand early hormonally regulated ES cell differentiation with potential application for significantly improving the yield of lineage-specific differentiation in vitro. This would greatly facilitate the development of ES cell technology for potential transplantation of "pure" cell populations into patients with diseases such as Parkinson disease, aplastic anemia, etc. In addition, the model proposed within this application predicts that histone deacetylase inhibitors such as valproic acid (a known teratogen) and TSA may greatly potentiate the teratogenic effects of environmental compounds by altering the very early histone acetylation pattern required for normal lineage-specific differentiation.

胚胎干细胞（ES）分化的最早阶段的分子方面仍然很差 明白我们的初步数据表明，整体组蛋白乙酰化可能是一个关键的第一步， 分化该提案的目标是确定是否全局组蛋白乙酰化和/或 甲基化发生在多次胚胎干细胞诱导分化的方法中，并建立了胚胎干细胞的甲基化模型。 使用组蛋白修饰的标准测定，这些组蛋白修饰发生的时间范围。我们 模型预测，这些组蛋白修饰的大部分可能发生在启动子区域， 利用新的CpG岛阵列来证实这一点。研究旨在确定全球是否 在从未分化的ES细胞状态退出期间发生的组蛋白修饰独特地不同于 激素信号对高度分化细胞诱导的基因特异性组蛋白修饰。的 第二个具体目标是探索导致这些独特的全球组蛋白的可能机制。 通过筛选ES细胞提取物的各种酶活性， 负责共价修饰历史。第三个具体目标是测试功能 组蛋白修饰在直接调节ES细胞分化中的意义。组蛋白脱乙酰酶 抑制剂，阿司他丁A（TSA），将与特定的生长因子结合使用，以增加速度， 定向于特定定向细胞命运的细胞的比例。此外，特定基因的过度表达 设计用于抑制ES中组蛋白乙酰转移酶活性或增加组蛋白脱乙酰酶活性的产品 评估细胞对ES细胞分化速率和总细胞命运定型的影响。的 拟议的研究旨在了解早期胚胎调控的ES细胞分化， 用于显著提高体外谱系特异性分化的产量。这将大大 促进ES细胞技术的发展，用于将“纯”细胞群移植到 帕金森病、再生障碍性贫血等疾病患者。此外， 在本申请中预测组蛋白脱乙酰酶抑制剂如丙戊酸（一种已知的致畸剂） 和TSA可能会大大加强环境化合物的致畸作用，通过改变非常 正常谱系特异性分化所需的早期组蛋白乙酰化模式。

项目成果

Chromatin context dominates estrogen regulation of pS2 gene expression.

• DOI: 10.1016/j.yexcr.2008.07.006
• 发表时间: 2008-09-10
• 期刊: Experimental cell research
• 影响因子: 3.7
• 作者: Oduro AK;Fritsch MK;Murdoch FE
• 通讯作者: Murdoch FE

Differentiation of murine embryonic stem cells induces progesterone receptor gene expression.

小鼠胚胎干细胞的分化诱导孕激素受体基因表达。

• DOI: 10.1016/j.yexcr.2005.09.005
• 发表时间: 2005
• 期刊: Experimental cell research.
• 作者: Sauter,CarleyN;McDermid,RebeccaL;Weinberg,AmyL;Greco,TamaraL;Xu,Xiaojie;Murdoch,FernE;Fritsch,MichaelK
• 通讯作者: Fritsch,MichaelK