Hematopoietic stem cells from human pluripotent stem cells and modeling of hemato

来自人类多能干细胞的造血干细胞和血液模型

• 批准号: 8748928
• 负责人: Sergei Doulatov
• 金额: $ 13.17万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 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 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8748928
• 关键词: Adult; Ascaridil; 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; Commit; Communities; Complex; Congenital Anemia; Dana-Farber Cancer Institute; Derivation procedure; Development; Development Plans; Diamond-Blackfan anemia; Disease; Doctor of Philosophy; Drug Design; Dysmyelopoietic Syndromes; Educational process of instructing; Embryonic Development; Engineering; Environment; Erythrocytes; Fostering; Funding; Generations; Genes; Globin; Goals; HOXA9 gene; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic Neoplasms; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; Hemoglobin; Heterogeneity; Hospitals; Human; In Vitro; Individual; Industry; Institutes; Institution; Israel; Knock-out; Lead; Leadership; Life; Malignant Neoplasms; Medical; Medical Research; 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 Factor; 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; disease phenotype; drug testing; fetal; in vivo; in vivo Model; 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; transcription factor

PROJECT SUMMARY 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 skills 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.

项目总结 血液系统是由罕见的造血干细胞(HSCs)维持的发育等级 扩展的自我更新和多谱系分化。因为它们有能力完全重建血液 在移植后，造血干细胞是一种非常有价值的治疗细胞类型。在我博士的博士培训期间。 约翰·迪克，我获得了专业知识，并为了解脐带血中的人类造血干细胞做出了贡献 (NAT免疫系统2010年；科学2011年)。在我与乔治·戴利博士的博士后培训中，我指导了我所有的 为生成造血干细胞和其他有价值的人类血细胞所做的努力和专业知识，如可输血 红细胞，来自患者诱导的多能干细胞(IPSCs)。我的工作证明了这一点的可行性 通过证明人类IPSC来源的血液前体可以转化(或重新指定)来实现这一重要目标 使用转录因子HOXA9、ERG和RORA(细胞干细胞)转化为可移植的多系祖细胞 单元格2013)。将祖细胞重新定向为干细胞的想法是一种很有前途的方法，但我们仍然 不能产生真正的造血干细胞。在我的研究提案的指导部分，我将专注于确定 长期多系移植中IPSCs转化为HSCs的转录因子组合 潜力。在独立阶段，我将研究这些调控网络在正常造血中的作用。 在体外和体内，重新指定的IPSC祖细胞是一种特别有效的红细胞来源。人类 红细胞在体内经历珠蛋白转换以表达成人血红蛋白，模拟胎儿到成人的珠蛋白 出生后发生的变化。出于这个原因，我已经开始使用IPSC对两种先天性贫血进行研究 来自钻石黑扇贫血和镰状细胞性贫血患者的品系。通过使用转录因子 将iPSC来源的祖细胞移植到小鼠身上，我将创建第一个人类血液疾病的体内模型， 询问潜在疾病机制并作为药物测试平台的目标(目标2a)。在 独立阶段，我将更深入地挖掘，使用IPSC因子系统来研究骨髓增生异常综合征 染色体5q缺失以分析缺失区间内的单个基因对疾病的贡献 病理生物学(目标2b)，这将确定靶向治疗的潜在途径。 戴利博士是国际上受人尊敬的干细胞生物学研究员，塞缪尔·E·勒克斯四世教授 血液学博士，波士顿儿童医院干细胞移植项目主任 医院(BCH)。戴利博士曾指导过36名工业界的主要研究人员和小组领导人，并 获得哈佛医学院颁发的A.Clifford Barger卓越导师奖。BCH是一种 著名的研究机构和儿科医学中心。BCH是医学和学术网络的一部分 包括哈佛医学院在内的大哈佛研究和医学界的机构， 达纳-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人提供了一个绝佳的机会 对科学家进行培训，开展高影响力的研究，并促进专业互动。戴利医生和我 制定了一份详细的职业发展计划，使我能够获得所需的技术、指导、 和领导才能。我还将得到一个干细胞生物学高级领导委员会的进一步指导： 伦纳德·宗、斯图尔特·奥金和戈登·凯勒。这次K99/R00奖项的支持将使我能够奉献我的 全力以赴执行这一雄心勃勃的项目，目标是在 指导阶段结束。 利用我在造血、多能干细胞和原生人体系统方面的专业知识，我将申请R00 在干细胞领域享有盛誉的顶尖研究所建立独立研究项目的资金 生物学。我相信，这项提议中概述的这种技能和创新研究计划的结合将 请允许我确立自己作为独立调查员的地位。我的职业目标是成为该领域的领军人物 开展多样化和协作性的研究计划，为 对基础生物学的洞察，同时为翻译、药物设计和基于细胞的研究创造真正的机会 治疗。