Characterization of embryonic stem cell-derived hematopoietic stem cells

描述(由申请人提供)： 提出了一项为期5年的计划，在该计划中，候选人将成为干细胞生物学的独立研究员，并进一步加深我们对胚胎干细胞生成的造血干细胞(ESC-HSC)的理解。在研究生院，这位候选人发展了成人干细胞方面的专业知识，专注于造血干细胞。作为一名博士后，她通过研究干细胞的分化潜力，初步探索CDX基因在干细胞造血发育中的作用，进一步加深了她的专业知识。这项工作目前正在审查中，准备出版。她目前的研究重点是ESC-HSC。尽管干细胞是新近从胚胎干细胞分化而来的，但其表型和功能尚不清楚，尤其是与体内干细胞的关系。候选人将通过阐明ESC-HSC的表型并将这些细胞在功能上进行比较，通过体内移植的HSC活性测试，以及通过全球基因表达的微阵列分析，在分子上，通过从体内发育的不同阶段纯化的小鼠的HSC，来检验ESC-HSC类似于体内发育不成熟的HSC隔室的假设。这项工作将在波士顿儿童医院血液/肿瘤科进行，由干细胞生物学的领军人物乔治·戴利博士指导。该部门拥有多个以干细胞研究成就著称的实验室(例如，Stuart Orkin博士和Len Zon博士)，并为推动该领域的发展营造了丰富的合作氛围。为了确保自己成为一名独立的研究人员并成功地完成拟议的工作，应聘者必须1)扩大她在与其研究相关的新领域的专业知识(例如发育生物学)，2)获得新的技术专业知识(例如胚胎解剖)，以及3)发展过渡到独立职位所需的技能(例如授权术)。为此，在广泛咨询了她的主要导师后，她制定了一个个性化的计划，包括说教和实践课程、研讨会和研讨会，以及一个由经验丰富的教员组成的委员会，他们将每半年与她会面一次，指导她的研究和职业发展。这项计划的执行，以及这项计划中概述的研究目标，将使应聘者能够发展专业知识，使自己成为ESC研究领域的独立和有竞争力的研究员。 公共卫生相关性：骨髓移植(BMT)被广泛用于治疗血液系统疾病。然而，许多患者缺乏捐赠者或必须使用非理想的捐赠者，导致危及生命的并发症。可移植细胞的替代来源将极大地扩大这种救命程序的使用范围，并将并发症降至最低。胚胎干细胞具有产生任何细胞类型的能力，代表着为骨髓移植寻找替代细胞来源的希望。这项提案中列出的工作将通过增加我们对从胚胎干细胞中分离出的细胞的了解来显著推进这一目标。

DESCRIPTION (provided by applicant): A 5-year plan is proposed in which the candidate will establish herself as an independent investigator in Stem Cell biology and further our understanding of hematopoietic stem cells generated by embryonic stem cells (ESC-HSC). In graduate school, the candidate developed an expertise in adult stem cells, focusing on HSC. As a postdoc, she furthered her expertise by researching the differentiative potential of ESC, initially exploring the role of the Cdx genes in hematopoietic development from ESC. This work is currently under review for publication. Her current research focuses on ESC-HSC. Although HSC were recently generated from ESC, nothing is known about their phenotype and function, especially in relation to in vivo HSCs. The candidate will test the hypothesis that ESC-HSC are akin to a developmentally immature in vivo HSC compartment by elucidating the phenotype of the ESC-HSC and comparing these cells functionally, by in vivo transplantation assays of HSC activity, and molecularly, via microarray analysis of global gene expression, to HSC purified from mice at different stages of in vivo development. This work will be conducted in the Division of Hematology/Oncology at Children's Hospital Boston under the mentorship of Dr. George Daley, a leader in stem cell biology. The Division is home to multiple laboratories renowned for their achievements in stem cell research (e.g. Drs. Stuart Orkin and Len Zon) and is a rich and collaborative atmosphere for advancing this field. To secure herself as an independent investigator and successfully complete the proposed work, the candidate must 1) expand her expertise in new fields pertinent to her research (e.g. Developmental Biology), 2) acquire new technical expertise (e.g. embryo dissection), and 3) develop the skills necessary to transition to an independent position (e.g. grantsmanship). Towards this end, after extensive consultation with her primary mentor, she has developed a personalized program composed of didactic and practical coursework, seminars and workshops, and a committee of experienced faculty who will meet with her semi-annually to guide her research and career development. The execution of this program, and the research aims outlined in this proposal, will allow the candidate to develop the expertise to establish herself as an independent and competitive investigator in ESC research. PUBLIC HEALTH RELEVANCE: Bone marrow transplantation (BMT) is extensively exploited for the treatment of hematopoietic maladies. Yet, many patients lack donors or must utilize non-ideal donors, resulting in life-threatening complications. Alternative sources of transplantable cells would greatly expand access to this life-saving procedure and minimize complications. Embryonic stem cells, with their capacity to generate any cell type, represent one hope for deriving an alternative cell source for BMT. The work out-lined in this proposal will significantly advance goal by increasing our understanding of cells isolated from embryonic stem cells.