Defining the molecular basis controlling hematopoietic stem cell symmetric and asymmetric divisions -supplement

定义控制造血干细胞对称和不对称分裂的分子基础 - 补充

• 批准号: 10765290
• 负责人: Hanzhi Luo
• 金额: $ 5.4万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10765290
• 关键词: Achievement; Affect; Bar Codes; Blood; Blood Cells; Cell Fate Control; Cells; Clinic; Data; Dedications; Dependence; Development Plans; Disease; Environment; Equilibrium; Goals; Hematological Disease; Hematopoiesis; Hematopoietic; Hematopoietic System; Hematopoietic stem cells; Homeostasis; In Vitro; Knowledge; Memorial Sloan-Kettering Cancer Center; Mentors; Methods; Modeling; Molecular; Persons; Pharmacology; Postdoctoral Fellow; Publishing; RNA; RNA methylation; RUNX1 gene; Reporting; Research; Role; Stress; System; Therapeutic; Training; Translating; Work; cancer stem cell; career development; hematopoietic stem cell fate; high throughput screening; induced pluripotent stem cell; leukemic stem cell; novel; novel therapeutics; programs; self-renewal; single-cell RNA sequencing; stem cell division; stem cell expansion; stem-like cell; stemness; success

PROJECT SUMMARY AND ABSTRACT Hematopoietic stem cells (HSCs) maintain blood homeostasis through a delicate balance in fate choices of self-renewal and commitment to differentiation. An imbalance in HSC fate choices is the key contributor to hematologic diseases that affect millions of people. HSC fate choices is maintained by coordinated symmetric self-renewal, symmetric commitment and asymmetric divisions. Thus, understanding the control of HSC division choices holds therapeutic potential for hematopoietic diseases. However, our knowledge on the control of HSC divisions is still lacking. Here, I aim to fill the gap by implementing novel systems and high-throughput assays to interrogate HSC fate choices. We recently revealed a unique role of m6A RNA methylation that is essential for HSC symmetric commitment, the division choice that is critical for the rapid replenishment of mature blood cells upon stress. Hence, I propose to leverage the m6A deficient HSCs as a novel system to elucidate the m6A-associated molecular programs in controlling HSC symmetric commitment. To uncover novel regulators in HSC fate choices, I aim to define the role of the understudied RBP NYNRIN, which is a stemness factor identified from my preliminary data and our recent AML-iPSC study, in controlling HSC divisions. I also aim to implement FATE-seq, a novel method involves HSC barcoding followed by in vitro division and single cell RNAseq that allows for a global assessment of HSC divisions. FATE-seq as an orthogonal system will identify novel regulators of HSC fate choices. The Specific Aims are: (1): Characterize the HSC-like intermediate state controlled by m6A RNA methylation. (2): Determine the molecular programs driven by m6A RNA methylation in controlling HSC symmetric commitment. (3): Uncover novel regulators in HSC symmetric and asymmetric fate decisions. Success of this work will define the novel regulators of HSC fate control and provide novel therapeutic opportunities for hematological diseases and stem cell expansion. Dr. Hanzhi Luo is currently a postdoc fellow in the Molecular Pharmacology program at the Memorial Sloan Kettering Cancer Center. Her initial achievements include uncovering m6A's role in HSC symmetric commitment, as well as revealing the RUNX1 dependency in leukemia stem cells with the AML-iPSC model. Both studies were recently published in Cell Reports. Her long-term goal is to establish a research team with an emphasis on molecular mechanisms of HSC fate choices with a strong commitment to translate basic sdiscoveries into the clinic. The proposed research will provide new training for Dr. Luo in hematopoiesis and RNA regulators. This work will be performed at the MSKCC, an exciting research environment with cutting- edge facilities. Dr. Luo will be mentored by Dr. Michael Kharas, a leading expert in studying RNA regulators in normal and malignant stem cells, and a dedicated supporter for young trainees. Together, this career development plan will help Dr. Luo establish her independent research lab at a research-oriented academic institute and become a leader in the field of hematopoiesis.

项目摘要和摘要 造血干细胞（HSC）通过命运选择的微妙平衡维持血液稳态 自我更新和对差异化的承诺。 HSC命运选择的不平衡是造成的关键因素 影响数百万人的血液学疾病。 HSC命运选择由协调的对称来维持 自我更新，对称的承诺和不对称分裂。因此，了解HSC的控制 分裂选择具有造血疾病的治疗潜力。但是，我们对控制的知识 HSC分区仍然缺乏。在这里，我的目标是通过实施新型系统和高通量来填补空白 询问HSC命运选择的测定法。我们最近揭示了M6A RNA甲基化的独特作用， HSC对称承诺至关重要，这对于快速补充至关重要的部门选择 压力后成熟的血细胞。因此，我建议利用M6A缺乏HSC作为一种新型系统 在控制HSC对称承诺时阐明了与M6A相关的分子程序。揭开 我的旨在定义研究不足的RBP Nynrin的作用，这是一个新的调节剂，这是 从我的初步数据和我们最近的AML-IPSC研究中确定的干性因子在控制HSC方面 部门。我还旨在实施命运赛，一种新颖的方法涉及HSC条形码，然后是体外 分裂和单细胞RNASEQ允许对HSC分区进行全球评估。命运seq作为一个 正交系统将确定HSC命运选择的新型调节因子。具体目的是：（1）：表征 由M6A RNA甲基化控制的HSC样中间状态。 （2）：确定分子程序 由M6A RNA甲基化驱动，以控制HSC对称承诺。 （3）：发现新颖的调节剂 HSC对称和不对称命运决策。这项工作的成功将定义HSC的新型监管机构 命运控制并为血液疾病和干细胞扩张提供新的治疗机会。 Hanzhi Luo博士目前是纪念Sloan的分子药理学计划的博士后研究员 Kettering癌症中心。她的最初成就包括揭示M6A在HSC对称中的作用 承诺，并通过AML-IPSC模型揭示白血病干细胞中的Runx1依赖性。 两项研究最近都发表在细胞报告中。她的长期目标是与 强调HSC命运选择的分子机制，他们坚定地承诺翻译基本 进入诊所的SD。拟议的研究将为Luo博士在造血和 RNA调节剂。这项工作将在MSKCC上进行，这是一个令人兴奋的研究环境，具有挑战 - 边缘设施。 Luo博士将由研究RNA调节器的主要专家Michael Kharas博士指导 在正常和恶性干细胞中，以及年轻学员的专用支持者。在一起，这个职业 发展计划将帮助Luo博士在以研究为导向的学术上建立她的独立研究实验室 研究所并成为造血领域的领导者。