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.

项目概要和摘要 造血干细胞 (HSC) 通过命运选择的微妙平衡来维持血液稳态 自我更新和致力于差异化。 HSC命运选择的不平衡是造成这种情况的关键因素 影响数百万人的血液疾病。 HSC 命运选择是通过协调对称来维持的 自我更新、对称承诺和不对称分裂。因此，了解 HSC 的控制 分支选择具有治疗造血系统疾病的潜力。然而，我们对控制的了解 HSC 部门仍然缺乏。在这里，我的目标是通过实施新颖的系统和高通量来填补空白 询问 HSC 命运选择的测定。我们最近揭示了 m6A RNA 甲基化的独特作用，即 对于 HSC 对称承诺至关重要，部门选择对于快速补充至关重要 应激时成熟的血细胞。因此，我建议利用 m6A 缺陷的 HSC 作为一种新颖的系统来 阐明控制 HSC 对称承诺的 m6A 相关分子程序。揭开 HSC 命运选择中的新型调节因子，我的目的是定义正在研究的 RBP NYNRIN 的作用，它是一个 根据我的初步数据和我们最近的 AML-iPSC 研究确定的干性因子，用于控制 HSC 部门。我还打算实现 FATE-seq，这是一种新颖的方法，涉及 HSC 条形码，然后进行体外 分裂和单细胞 RNAseq，可对 HSC 分裂进行全局评估。 FATE-seq 作为 正交系统将识别 HSC 命运选择的新调节因子。具体目标是： (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 甲基化驱动控制 HSC 对称承诺。 (3)：发现新的监管机构 HSC 对称和不对称命运决定。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增提供新的治疗机会。 罗瀚志博士目前是纪念斯隆管理学院分子药理学项目的博士后研究员 凯特林癌症中心。她最初的成就包括揭示 m6A 在 HSC 对称性中的作用 承诺，以及通过 AML-iPSC 模型揭示白血病干细胞中的 RUNX1 依赖性。 这两项研究最近发表在《细胞报告》上。她的长期目标是建立一个研究团队 强调 HSC 命运选择的分子机制，并坚定致力于转化基本原理 s发现进入临床。拟议的研究将为罗博士提供造血和免疫学方面的新训练。 RNA 调节剂。这项工作将在 MSKCC 进行，这是一个令人兴奋的研究环境，具有尖端技术 边缘设施。罗博士将接受RNA调节因子研究领域领先专家Michael Kharas博士的指导 正常和恶性干细胞，以及年轻学员的忠实支持者。一起，这个职业 发展计划将帮助罗博士在一个以研究为导向的学术机构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。