Aberrant glycolysis as a driver of mutant HSPC expansion in clonal hematopoiesis

异常糖酵解是克隆造血中突变型 HSPC 扩增的驱动因素

• 批准号: 10729107
• 负责人: Eric M Pietras
• 金额: $ 45.69万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10729107
• 关键词: Address; Aging; Biological Assay; Bone Marrow; Cardiovascular Diseases; Cells; Cellular Metabolic Process; Chronic; Clinical; Clonal Expansion; Clone Cells; DNMT3a; Data; Disease; Elderly; Evolution; Exhibits; Fire - disasters; Flow Cytometry; Genes; Glucose; Glycolysis; Goals; HIF1A gene; Health; Heart; Hematologic Neoplasms; Hematopoiesis; Hematopoietic; Hematopoietic stem cells; High Prevalence; Homeostasis; Human; Hyperactivity; Hypoxia Inducible Factor; Image; Immune; Individual; Inflammasome; Inflammation; Inflammatory; Investigation; Link; Mass Spectrum Analysis; Mediating; Metabolic; Metabolism; Modeling; Molecular Genetics; Morbidity - disease rate; Mus; Mutation; Myelogenous; Myeloid Cells; Myeloproliferative disease; Oral Administration; Pathogenicity; Pathway interactions; Phenotype; Prevalence; Process; Production; Resolution; Risk; Role; Signal Transduction; Smoking History; Therapeutic; Therapeutically Targetable; Tissues; Work; chronic inflammatory disease; comorbidity; cytokine; experimental study; fitness; genetic approach; genotoxicity; human old age (65+); improved; in vivo; inhibitor; innovation; metabolic phenotype; mortality; mouse model; mutant; novel strategies; overexpression; pharmacologic; prevent; therapeutic target; transcription factor; uptake

PROJECT SUMMARY / ABSTRACT The objective of this proposal is to identify new approaches for targeting clonal hematopoiesis (CH). CH is characterized by selective expansion of hematopoietic stem and progenitor cell (HSPC) clones harboring mutations in genes such as TET2 and DNMT3A. These HSPC in turn produce pathogenic mutant myeloid cells that can contribute to several aging-related co-morbidities including cardiovascular disease (CVD) and all-cause mortality. An expanded mutant HSPC pool may also contribute to increased risk of hematological malignancies. CH prevalence is significantly elevated in the elderly and individuals with prior genotoxic exposures, smoking history, and/or chronic inflammatory disease. These conditions are associated with chronically perturbed physiological homeostasis, characterized by hyper-inflammation. Understanding the mechanism(s) promoting the selective expansion of mutant HSPC is crucial for prioritizing therapeutic targets that can suppress CH. A central premise of our application is that that CH arises from a targetable interplay between inflammatory signals and altered metabolic programming that supports the energetic needs and thereby the preferential expansion of CH HSPC. Using the mouse as a model representing key features of human CH, our preliminary data show that CH HSPC exhibit increased levels of the transcription factor Hif-1 aberrant glycolytic metabolism and increased ATP production relative to wild-type HSPC. We find that inflammatory cytokines, particularly IL-1 strongly potentiates CH HSPC expansion, glycolytic metabolism and Hif-1 activity. Strikingly, treatment of CH mice with OLT-1177, an NLRP3 inflammasome inhibitor that prevents cleavage and activation of IL-1 potently suppresses CH. We hypothesize that CH is the result of an interdependent mechanism in which NLRP3-mediated IL-1 production potentiates Hif-1 and downstream glycolytic activity to support CH HSPC expansion. We propose that NLRP3 inhibition disrupts this circuit, limiting expansion of CH HSPC. To address the mechanism, we propose two Specific Aims: 1) we will characterize the metabolic features of Tet2/, Tet2+/ and Dnmt3aR878H/+ CH HSPC using in vivo mass spectrometry- and flow cytometry-based analyses of metabolism and glucose flux. We will also identify the extent to which CH HSPC rely upon Hif-1 and glycolysis for their energetic needs; 2) we will evaluate the requirement for NLRP3 in promoting aberrant glycolytic activity and/or preferential expansion of CH HSPC. We will use molecular genetics approaches to assess the role and mechanism of Hif-1 and NLRP3 in regulating CH HSPC metabolism and promoting their expansion in vivo. Using our non-conditioned adoptive BM transfer mouse model of CH, we will validate our mechanism using pharmacological inhibition of NLRP3 with OLT-1177 and establish whether NLRP3 blockade suppresses HSPC expansion, aberrant metabolic activity and accumulation of inflammatory immune cells in the heart and other tissues. This work has the potential to re-frame CH as a therapeutically targetable metabolic phenotype, significantly improving health among groups with high prevalence of CH such as the elderly.

项目概要/摘要 该提案的目的是确定针对克隆造血 (CH) 的新方法。 其特点是选择性扩增含有造血干细胞和祖细胞 (HSPC) 克隆 TET2 和 DNMT3A 等基因突变会产生致病性突变骨髓细胞。 可能导致多种与衰老相关的并发症，包括心血管疾病 (CVD) 和全因疾病 突变 HSPC 池的扩大也可能导致血液恶性肿瘤的风险增加。 老年人和曾接触过遗传毒性物质、吸烟的人的 CH 患病率显着升高 病史和/或慢性炎症性疾病与长期紊乱有关。 以过度炎症为特征的生理稳态。了解促进机制。 突变体 HSPC 的选择性扩增对于优先考虑可抑制 CH 的治疗靶点至关重要。 我们应用的一个核心前提是，CH 是由炎症之间的可靶向相互作用引起的。 信号和改变代谢程序，支持能量需求，从而优先 我们初步使用小鼠作为代表人类 CH 关键特征的模型进行了 CH HSPC 的扩展。 数据显示，CH HSPC 的转录因子 Hif-1 异常糖酵解水平升高 相对于野生型 HSPC，我们发现炎症细胞因子的代谢和 ATP 产量增加。 尤其是 IL-1 强烈增强 CH HSPC 扩增、糖酵解代谢和 Hif-1 活性。 使用 OLT-1177 治疗 CH 小鼠，OLT-1177 是一种 NLRP3 炎症小体抑制剂，可防止裂解和激活 IL-1 有效抑制 CH 我们认为 CH 是一种相互依赖机制的结果，其中 NLRP3 介导的 IL-1 产生增强 Hif-1 和下游糖酵解活性以支持 CH HSPC 我们认为 NLRP3 抑制会破坏该回路，从而限制 CH HSPC 的扩展。 为了解决这一机制，我们提出了两个具体目标：1）我们将表征 使用基于体内质谱和流式细胞术的 Tet2/、Tet2+/ 和 Dnmt3aR878H/+ CH HSPC 我们还将确定 CH HSPC 对 Hif-1 的依赖程度。 和糖酵解以满足其能量需求；2) 我们将评估 NLRP3 在促进异常方面的需求； 我们将使用分子遗传学方法来研究 CH HSPC 的糖酵解活性和/或优先扩增。 评估Hif-1α和NLRP3在调节CH HSPC代谢并促进其代谢中的作用和机制 使用我们的非条件过继 BM 移植小鼠模型，我们将验证我们的体内扩增。 使用 OLT-1177 对 NLRP3 进行药理抑制的机制，并确定 NLRP3 是否被阻断 抑制 HSPC 扩增、异常代谢活动和炎症免疫细胞的积累 这项工作有可能将 CH 重新定义为一种可治疗的代谢靶点。 表型，显着改善 CH 患病率高的群体（例如老年人）的健康状况。