Clonal tracking and molecular characterization of hematopoiesis under stress

应激条件下造血的克隆追踪和分子特征

• 批准号: 10413504
• 负责人: David T Scadden
• 金额: $ 5.04万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10413504
• 关键词: Address; Age; Animal Model; Behavior; Blood Cells; Blood Flow Cytometry; Bone Marrow; CRISPR screen; Cells; Characteristics; Clonal Expansion; Clonal Hematopoietic Stem Cell; Clone Cells; Collection; Data; Development; Disadvantaged; Disease; Dysmyelopoietic Syndromes; Engineering; Epigenetic Process; Equilibrium; Fluorescence; Frequencies; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genetic; Genotoxic Stress; Goals; Habits; Hematological Disease; Hematology; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; Heterogeneity; Human; Individual; Inflammatory; Intervention; Kinetics; Label; Laboratories; Lesion; Lymphoid; Methods; Molecular; Molecular Analysis; Molecular Profiling; Mus; Mutation; Myelogenous; Neoplasms; Normal Cell; Phenotype; Population; Production; Radiation; Regulator Genes; Research; Resolution; Retrotransposon; Risk; Stimulus; Stress; System; Techniques; Testing; Time; Transplantation; Zebrafish; base; collaborative approach; comparative; genotoxicity; improved; in vivo; insight; mortality; mutant; novel; offspring; premalignant; primitive cell; progenitor; response; small hairpin RNA; stem cells

RESEARCH SUMMARY Hematopoiesis is now recognized as the integrated activity of multiple, largely progenitor clones with heterogeneous behavior. Clonal complexity is thought to diminish with age and human studies suggest that oligoclonal hematopoiesis with disease associated mutations is common. The presence of such clones incurs a substantial risk of hematologic neoplasia and all cause mortality. The overall goal of this proposal is to understand the molecular drivers of clonal behavior under stress conditions and to test whether modifying those drivers is capable of changing the relative competitive balance of normal and abnormal, risk bearing clones in animal models. We will use techniques of fluouresence clonal tracking in the mouse developed by us and compare these with retrotransposon clonal marking techniques in the mouse of Carmago and fluorescence clonal tracking techniques in the zebrafish developed by Zon. Further, we will characterize genetic characteristics of clones at single cell resolution using techniques of Tenen and define ncRNA candidate molecular drivers with Tenen. Finally, we will target epigenetic modifiers defined by Orkin, Tenen and Zon to test the impact on clonal competition between normal and mutant clones in vivo. Combined these complementary and collaborative approaches will point to methods for altering competitive relationships in hematopoiesis in favor of normal cells.

研究综述 造血现在被认为是多个，主要是祖细胞克隆的综合活动，具有 异质行为。克隆的复杂性被认为随着年龄的增长而减少，人类研究表明 带有疾病相关突变的寡克隆性造血是常见的。这种克隆的存在会招致 有很大的血液肿瘤风险，所有这些都会导致死亡。这项提案的总体目标是 了解应激条件下克隆行为的分子驱动因素，并测试是否修改 这些驱动力能够改变正常和非正常、承担风险的相对竞争平衡 动物模型中的克隆。我们将在我们开发的小鼠中使用荧光克隆跟踪技术 并将其与Carmago和Carmago小鼠的逆转录转座子克隆标记技术进行比较。 Zon开发的斑马鱼荧光克隆示踪技术。此外，我们还将描述 用Tenen和Defined ncRNA技术研究单细胞分辨率克隆的遗传特征 Tenen的候选分子驱动器。最后，我们将针对由田纳西州Orkin定义的表观遗传修饰 和Zon在体内测试对正常克隆和突变克隆之间克隆竞争的影响。将这些组合在一起 互补和协作的方法将指向改变竞争关系的方法 有利于正常细胞的造血。