Origin of HSC from Embryonic Stem Cells

胚胎干细胞 HSC 的起源

基本信息

批准号：
7264502
负责人：
ELIAS T. ZAMBIDIS
金额：
$ 13.31万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-31 至 2009-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7264502
关键词：
Address Basic Science Biomedical Research Birds Cell Differentiation process Cellular biology Clinical Complementary DNA Condition Derivation procedure Development Embryo Endothelium Fetal Thymic Organ Culture Future Gene Targeting Genes Genetic Goals Helix-Turn-Helix Motifs Hematopoiesis Hematopoietic Hematopoietic Stem Cell Transplantation Hematopoietic stem cells Human Knock-in Mouse LacZ Genes Ligands Lymph Mentored Clinical Scientist Development Award (K08)Methods Modeling Modification Molecular Mus Nature Notch Signaling Pathway Nucleic Acid Regulatory Sequences Numbers Pathway interactions Play Reporter Research Research Proposals Research Training Role Signal Transduction Signal Transduction Pathway Stem cell transplant Stem cells T-Cell Development T-Lymphocyte T-cell acute lymphocytic leukemia 1 protein Training Programs Zebrafish career embryonic stem cell genetic analysis hematopoietic tissue homologous recombination human embryonic stem cell human stem cells interest notch protein progenitor programs success tool

项目摘要

DESCRIPTION (provided by applicant): The nature of this Mentored Clinical Scientist Development Award (K08) pertains to the understanding of human pluripotent and hematopoietic stem cells. This proposed 5-year research training program will advance the study of human developmental hematopoiesis with one of the most important new tools in biomedical research: the human embryonic stem cell (hES). My long-term interest is in advancement of clinical stem cell transplantation, and I intend to devote a large part of my career to basic research in stem cell biology. This proposal will focus on the derivation, culture, and expansion of primitive human hematopoietic stem cells (HSC) from hES. The strategy for identifying early human HSC will rely on the general hypothesis that a "hemangioblast" (bipotential progenitor of HSC and endothelium) and/or a "hemogenic endothelial" precursor can be physically isolated and characterized from differentiating hES using experimental approaches similar to those described in other species. These hES-derived progenitors will be used for subsequent studies in human lympho-hematopoietic development. The research program will initially focus on expanding, purifying, and characterizing hES-derived hemangioblasts. Subsequent studies will probe the mechanistic role SCL/TAL1, a master regulator for initiating hematopoiesis, plays on the formation of the human hemangioblast. A reporter cDNA will be gene-targeted to the endogenous SCL locus regulatory region ("knock-in") with modified methods of homologous recombination, and primitive SCL-expressing hemato-endothelial progenitors will be purified and expanded for further phenotypic and genetic analysis. Primitive HSC will ultimately be cultured on stromal lines and/or fetal thymic organ cultures for the derivation of a model for human T-lymphocyte development. The role of Notch pathway signaling in hES-drived lymph-hematopoietic tissue will be studied as a long-term goal. The success of these goals will lay the groundwork for characterizing human stem cells not available by any other means.

描述（由申请人提供）：该指导临床科学家发展奖（K08）的性质涉及对人类多能和造血干细胞的理解。这项拟议的为期 5 年的研究培训计划将利用生物医学研究中最重要的新工具之一：人类胚胎干细胞 (hES) 来推进人类发育造血的研究。我的长期兴趣是临床干细胞移植的进步，我打算将我职业生涯的很大一部分投入到干细胞生物学的基础研究上。该提案将重点关注从 hES 衍生、培养和扩增原始人类造血干细胞 (HSC)。鉴定早期人类 HSC 的策略将依赖于这样一个一般假设：“成血管细胞”（HSC 和内皮细胞的双能祖细胞）和/或“造血内皮细胞”前体可以通过使用类似于这些实验方法的实验方法从分化 hES 中物理分离和表征。在其他物种中也有描述。这些 hES 衍生的祖细胞将用于人类淋巴造血发育的后续研究。该研究项目最初将集中于扩展、纯化和表征 hES 衍生的成血管细胞。后续研究将探讨 SCL/TAL1（造血起始的主要调节因子）在人类成血管细胞形成中的机制作用。报告基因 cDNA 将通过同源重组的改良方法基因靶向内源 SCL 基因座调节区（“敲入”），并且将纯化和扩展表达原始 SCL 的血液内皮祖细胞，以进行进一步的表型和遗传分析。原始HSC最终将在基质系和/或胎儿胸腺器官培养物上培养，以衍生人类T淋巴细胞发育模型。 Notch 通路信号在 hES 驱动的淋巴造血组织中的作用将作为长期目标进行研究。这些目标的成功将为表征人类干细胞奠定基础，而这是其他任何方法都无法实现的。