Hematopoietic stem cells from human pluripotent stem cells and modeling of blood disorders

来自人类多能干细胞的造血干细胞和血液疾病模型

• 批准号: 9335939
• 负责人: Sergei Doulatov
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9335939
• 关键词: Adult; Alpha Cell; Award; Biochemical; Biological; Biology; Birth; Blood; Blood Cells; Blood Platelets; Bone Marrow; Boston; Candidate Disease Gene; Cell Line; Cell Therapy; Cell model; Cells; Childhood; Chromosomes; Clinical; Communities; Complex; Congenital Anemia; Dana-Farber Cancer Institute; Derivation procedure; Development; Development Plans; Diamond-Blackfan anemia; Disease; Doctor of Philosophy; Drug Design; Dysmyelopoietic Syndromes; Embryonic Development; Engineering; Environment; Erythrocytes; Fostering; Funding; Generations; Genes; Genetic Transcription; Globin; Goals; HOXA9 gene; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic stem cells; Hemoglobin; Hospitals; Human; In Vitro; Individual; Industry; Institutes; Institution; International; Israel; Knock-out; Leadership; Life; Malignant Neoplasms; Medical; Medical center; Mentors; Mentorship; Methods; Modeling; Mus; Paper; Patients; Pediatric Hospitals; Phase; Phenotype; Pluripotent Stem Cells; Principal Investigator; Protocols documentation; RORA gene; Research; Research Institute; Research Personnel; Research Proposals; Role; Science; Scientist; Sickle Cell Anemia; Somatic Cell; Source; Specific qualifier value; Stem cell transplant; Stem cells; System; Testing; Therapeutic; Time; Tissues; Training; Transcriptional Regulation; Translations; Transplantation; Umbilical Cord Blood; Umbilical Cord Blood Transplantation; Woman; Work; Xenograft procedure; career; career development; cell type; cellular engineering; chromosome 5q loss; curative treatments; disease heterogeneity; disease phenotype; drug testing; fetal; human pluripotent stem cell; in vivo; in vivo Model; individual patient; induced pluripotent stem cell; innovation; insight; interest; medical schools; mouse model; novel; novel strategies; post-doctoral training; professor; progenitor; programs; reconstitution; self-renewal; skills; stem cell biology; targeted treatment; transcription factor

DESCRIPTION (provided by applicant): The blood system is a developmental hierarchy maintained by rare hematopoietic stem cells (HSCs) capable of extended self-renewal and multilineage differentiation. Because of their ability to fully reconstitute the blood system upon transplantation, HSCs are a highly valued therapeutic cell type. During my Ph.D. training with Dr. John Dick, I have gained expertise and contributed to understanding of human HSCs from umbilical cord blood (Nat Immunol 2010; Science 2011). In my postdoctoral training with Dr. George Daley, I have directed all my efforts and expertise to the generation of HSCs and other valuable human blood cells, such as transfusable red blood cells, from patient induced-pluripotent stem cells (iPSCs). My work has demonstrated the feasibility of this important goal by showing that human iPSC-derived blood precursors can be converted (or respecified) into transplantable multilineage progenitors using transcription factors: HOXA9, ERG, and RORA (Cell Stem Cell 2013). The idea of respecifying progenitors into stem cells is a promising approach, however we are still short of generating true HSCs. In the mentored part of my research proposal, I will focus on identifying combinations of transcription factors that respecify iPSCs into HSCs with long-term multilineage transplantation potential. In the independent phase, I will study the role of these regulatory networks in normal hematopoiesis. Respecified iPSC progenitors are a particularly potent source of red blood cells in vitro and in vivo. Human erythrocytes undergo globin switching in vivo to express adult hemoglobin mimicking the fetal-to-adult globin switch that occurs after birth. For this reason, I have initiated the study of two congenital anemias using iPSC lines from patients with Diamond Blackfan anemia and sickle cell anemia. By using the transcription factors to engraft iPSC-derived progenitors in mice, I will create the first in vivo models of human blood disorders, with the goal of interrogating the underlying disease mechanisms and as a platform for drug testing (Aim 2a). In the independent phase, I will delve deeper, using the iPSC factor system to study myelodysplastic syndromes with a chromosome 5q deletion to dissect the contribution of individual genes within the deletion interval to disease pathobiology (Aim 2b), which will identify potential avenues for targeted therapies. Dr. Daley is an internationally respected investigator in stem cell biology, the Samuel E. Lux IV Professor of Hematology, and serves as the Director of the Stem Cell Transplantation Program at Boston Children's Hospital (BCH). Dr. Daley has mentored 36 principal investigators and group leaders in industry, and has received the A. Clifford Barger Excellence in Mentoring Award from Harvard Medical School. BCH is a prestigious research institute and a pediatric medical center. BCH is part of a network of medical and academic institutions within the greater Harvard research and medical community, that includes Harvard Medical School, Dana-Farber Cancer Institute, Brigham and Women's Hospital, Beth Israel Deaconess Medical Center, Harvard Stem Cell Institute, and others. This rich environment provides a superb opportunity for a young scientists to train, carry out high impact research, and foster professional interactions. Dr. Daley and I have developed a detailed career development plan that will allow me to acquire the needed technical, mentorship, and leadership skills. I will be further guided by a committee of senior leaders in stem cell biology: Drs. Leonard Zon, Stuart Orkin, and Gordon Keller. The support of this K99/R00 award will allow me to dedicate my full energies to carry out this ambitious project with the goal of producing several high-impact papers by the end of mentored phase. Using my expertise in hematopoiesis, pluripotent stem cells, and primary human systems, I will apply for R00 funds to establish an independent research program at a top institute with a reputation as a leader in stem cell biology. I believe that this combination of sklls and an innovative research plan outlined in this proposal will allow me to establish myself as an independent investigator. My career goal is to become a leader in the field of stem cell biology, to carry out a diverse and collaborative research program that provides fundamental insights into basic biology, while creating real opportunities for translation, drug design, and cell-based therapies.

描述（由申请人提供）：血液系统是由能够延长自我更新和多谱系分化的罕见造血干细胞（HSC）维持的发育层次。由于它们在移植后完全重建血液系统的能力，HSC是一种非常有价值的治疗性细胞类型。在我读博士的时候。通过与John Dick博士的培训，我获得了专业知识，并为了解来自脐带血的人类HSC做出了贡献（Nat Immunol 2010; Science 2011）。在我与乔治戴利博士的博士后培训中，我将我所有的努力和专业知识用于从患者诱导的多能干细胞（iPSC）中产生HSC和其他有价值的人类血细胞，如可再生红细胞。我的工作已经证明了这一重要目标的可行性，通过显示人类iPSC衍生的血液前体可以使用转录因子转换（或重新指定）为可移植的多系祖细胞：HOXA 9，ERG和RORA（Cell Stem Cell 2013）。将祖细胞重新指定为干细胞的想法是一种有前途的方法，但我们仍然缺乏真正的HSC。在我的研究计划的指导部分，我将重点放在识别转录因子的组合， 将iPSC转化为具有长期多谱系移植潜力的HSC。在独立阶段，我将研究这些调节网络在正常造血中的作用。重新指定的iPSC祖细胞是体外和体内红细胞的特别有效的来源。人类红细胞在体内经历珠蛋白转换以表达成人血红蛋白，模仿出生后发生的胎儿至成人珠蛋白转换。出于这个原因，我已经开始使用来自Diamond Blackfan贫血和镰状细胞贫血患者的iPSC系对两种先天性贫血进行研究。通过使用转录因子在小鼠中移植iPSC衍生的祖细胞，我将 创建人类血液疾病的第一个体内模型，目的是探究潜在的疾病机制，并作为药物测试的平台（目标2a）。在独立阶段，我将深入研究，使用iPSC因子系统研究染色体5 q缺失的骨髓增生异常综合征，以剖析缺失区间内单个基因对疾病病理学的贡献（目标2b），这将确定靶向治疗的潜在途径。戴利博士是一位国际上受人尊敬的干细胞生物学研究者。勒克斯IV血液学教授，并担任波士顿儿童医院（BCH）干细胞移植项目主任。戴利博士指导了36名主要研究人员和行业领导者，并获得了A。获得哈佛医学院的Clifford Barger优秀指导奖。BCH是一个著名的研究机构和儿科医疗中心。BCH是更大的哈佛研究和医学界的医疗和学术机构网络的一部分，包括哈佛医学院，丹娜法伯癌症研究所，布里格姆妇女医院，贝丝以色列女执事医疗中心，哈佛干细胞研究所等。这种丰富的环境为年轻科学家提供了培训，开展高影响力研究和促进专业互动的绝佳机会。戴利博士和我已经制定了一个详细的职业发展计划，这将使我获得所需的技术，指导和领导技能。我将进一步接受干细胞生物学高级领导人委员会的指导：伦纳德宗博士、斯图尔特奥金博士和戈登凯勒博士。这个K99/R 00奖的支持将使我能够全身心投入到这个雄心勃勃的项目中，目标是在指导阶段结束时产生几篇高影响力的论文。利用我在造血，多能干细胞和初级人类系统方面的专业知识，我将申请R 00资金，在一个拥有干细胞生物学领导者声誉的顶级研究所建立一个独立的研究项目。我相信，这份建议书中概述的技能和创新研究计划的结合将使我成为一名独立的研究者。我的职业目标是成为干细胞生物学领域的领导者，开展多样化的合作研究计划，为基础生物学提供基本见解，同时为翻译，药物设计和基于细胞的疗法创造真实的机会。