Hematopoietic/erythropoietic differentiation of human embryonic stem cells

人胚胎干细胞的造血/红细胞分化

• 批准号: 7892600
• 负责人: Kai-Hsin Chang
• 金额: $ 12.71万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7892600
• 关键词: Address; Adult; Advisory Committees; BFU-E; Bone Marrow; CD34 gene; Cell fusion; Cells; Coculture Techniques; Committee Members; Complex; Development; Ectopic Expression; Educational process of instructing; Elements; Embryo; Environment; Erythroid; Erythroid Cells; Erythropoiesis; Ethical Issues; Event; Fetal Liver; Fetal Tissues; Future; Gene Expression; Generations; Genes; Globin; Goals; Hematopoiesis; Hematopoietic; Hepatocyte; Homeobox; Human; Laboratories; Learning; Mentors; Methods; Molecular; Molecular Biology; Morphology; Mus; Nature; Pattern; Population; Principal Investigator; Regulation; Regulator Genes; Reporting; Research; Research Personnel; Spumavirus; Stromal Cells; Surface; Techniques; Thalia (angiosperm); Time; Training; Undifferentiated; Work; base; beta Globin; chromatin modification; design; erythroid differentiation; experience; fetal; gamma Globin; human embryonic stem cell; insight; overexpression; progenitor; programs; tool; transcription factor; vector

DESCRIPTION (provided by applicant): This proposal is designed to provide the Principal Investigator, Kai-Hsin Chang, with a training experience in the fields of molecular biology to expand the characterization of the commitment and differentiation of hematopoietic/erythroid cells from human embryonic stem cells (hESCs) and to study the expression of beta- locus globin genes at both cellular and molecular levels. The proposed studies are based on the observation that while hESCs can be induced to undergo hematopoietic and erythroid differentiation to help delineate the mechanisms underlying early events of hematopoiesis, the efficiency of generating hematopoietic/erythroid progenitors is low. Furthermore, unlike their fetal liver counterparts that mainly express fetal gamma globin, hESC-derived fetal-like definitive erythroid cells coexpress high levels of embryonic (epsilon) and fetal (gamma) globins with little adult beta globin. Thus, hESC-derived erythroid cells represent a valuable tool to study the activation and silencing of the beta-locus globin genes. The following specific aims are proposed: 1) To study the effect of ectopic expression of homeobox (Hox) transcription factor HoxB4 on the hematopoietic/erythroid differentiation of hESCs. 2) To characterize the cis- and trans-acting elements associated with the unique beta-locus globin expression pattern of hESC-derived erythroid cells. 3) To study the modulation of beta-locus globin gene expression by manipulation of intracellular transcriptional environment via cell fusion. While the applicant's short-term goal is to learn the molecular techniques to investigate hematopoietic/erythropoietic differentiation of hESCs and to study the regulatory mechanisms governing the globin expression. Her long-term goal is to develop an independent research program devoted to the study of erythroid cells, which will contribute to the development of althernative therapy for treating hemoglobinpathies. Dr. Thalia Papayannopoulou's laboratory has an outstanding record of studying the globin regulation using cellular and molecular approaches and thus will provide the applicant with an appropriate and stimulating teaching environment. The applicant will also benefit from the expertise of several other investigators nearby, including Dr. George Stamatoyannopoulos, the co-mentor, and Drs. David Russell, and Xiangdong Fang, the advisory committee members, necessary to accomplish the goal of present application and future objectives.

描述（由申请人提供）： 该提案旨在为主要研究者Kai-Hsin Chang提供分子生物学领域的培训经验，以扩展人胚胎干细胞（hESC）造血/红系细胞定型和分化的表征，并研究β-位点珠蛋白基因在细胞和分子水平上的表达。提出的研究是基于这样的观察，即虽然hESC可以被诱导经历造血和红细胞分化以帮助描述造血早期事件的潜在机制，但产生造血/红细胞祖细胞的效率很低。此外，与主要表达胎儿γ球蛋白的胎儿肝对应物不同，hESC衍生的胎儿样定形红系细胞共表达高水平的胚胎（γ）和胎儿（γ）球蛋白，几乎没有成人β球蛋白。因此，hESC衍生的红系细胞代表了研究β基因座珠蛋白基因的激活和沉默的有价值的工具。本研究的具体目的如下：1）研究同源异型盒（homeobox，Hox）转录因子HoxB4的异位表达对hESC造血/红系分化的影响。2)表征与hESC衍生的红系细胞的独特β位点珠蛋白表达模式相关的顺式和反式作用元件。3)研究通过细胞融合调控细胞内转录环境对β位点珠蛋白基因表达的调控。而申请人的短期目标是学习分子技术来研究hESCs的造血/红细胞生成分化，并研究控制球蛋白表达的调节机制。她的长期目标是开发一个独立的研究计划，致力于红系细胞的研究，这将有助于开发治疗血红蛋白病的替代疗法。Thalia Papayannopoulou博士的实验室在使用细胞和分子方法研究珠蛋白调节方面有着出色的记录，因此将为申请人提供适当和刺激的教学环境。申请人还将受益于附近其他几位研究者的专业知识，包括共同导师乔治斯塔马托扬诺普洛斯博士和顾问委员会成员大卫罗素博士和方向东博士，这些都是实现当前申请目标和未来目标所必需的。