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

基本信息

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
项目类别：
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

项目摘要

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.

研究概要造血现在被认为是多个主要祖细胞克隆的整合活动异质行为。人们认为克隆复杂性会随着年龄的增长而降低，人类研究表明具有疾病相关突变的寡克隆造血很常见。此类克隆的存在会导致血液肿瘤和全因死亡的巨大风险。该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，并测试是否会改变这些驱动因素能够改变正常与异常、风险承担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追踪技术并将这些与 Carmago 小鼠中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追踪技术。进一步，我们将表征使用 Tenen 技术在单细胞分辨率下克隆的遗传特征并定义 ncRNA Tenen 的候选分子驱动器。最后，我们将针对 Orkin、Tenen 定义的表观遗传修饰因子和 Zon 来测试体内正常克隆和突变克隆之间克隆竞争的影响。结合这些互补和协作方法将指出改变竞争关系的方法有利于正常细胞的造血作用。