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.

项目摘要/摘要 本提案的目的是确定新的方法，针对克隆造血（CH）。CH是 其特征在于选择性扩增具有以下特征的造血干细胞和祖细胞（HSPC）克隆： TET 2和DNMT 3A等基因突变。这些HSPC反过来又产生致病突变的髓样细胞 这可能导致几种与衰老相关的合并症，包括心血管疾病（CVD）和各种原因 mortality.扩增的突变体HSPC合并液也可能导致血液学恶性肿瘤风险增加。 CH患病率在老年人和既往有遗传毒性暴露、吸烟的个体中显著升高 病史和/或慢性炎性疾病。这些情况与慢性扰动有关 生理稳态，其特征在于过度炎症。了解促进 突变体HSPC的选择性扩增对于优先选择能够抑制CH的治疗靶点是至关重要的。 我们的应用的一个中心前提是，CH是由炎症之间的靶向相互作用引起的。 信号和改变的代谢程序，支持能量的需要，从而优先 CH HSPC的扩展。使用小鼠作为代表人类CH关键特征的模型， 数据显示，CH HSPC表现出转录因子Hif-1水平增加，从而导致糖酵解异常， 相对于野生型HSPC，HSPC的代谢和ATP产生增加。我们发现炎症细胞因子， 特别是IL-1 β强烈增强CH HSPC扩增、糖酵解代谢和Hif-1 β活性。引人注目的是， 用OLT-1177治疗CH小鼠，OLT-1177是一种阻止裂解和活化的NLRP 3炎性体抑制剂 我们假设CH是一种相互依赖的机制的结果， NLRP 3介导的IL-1 β产生增强Hif-1 β和下游糖酵解活性以支持CH HSPC 扩张.我们认为NLRP 3抑制破坏了这一回路，限制了CH HSPC的扩增。 为了解决这一机制，我们提出了两个具体的目标：1）我们将表征的代谢特点， 使用基于体内质谱和流式细胞术的Tet 2 α/β、Tet 2 +/β和Dnmt 3aR 878 H/+ CH HSPC 代谢和葡萄糖通量的分析。我们还将确定CH HSPC依赖Hif-1的程度 2）我们将评估NLRP 3在促进异常糖酵解中的需求， 糖酵解活性和/或CH HSPC的优先扩增。我们将使用分子遗传学方法， 评估HIF-1 α和NLRP 3在调节CH HSPC代谢和促进其代谢中的作用和机制。 体内扩增使用我们的CH的非条件过继BM转移小鼠模型，我们将验证我们的 使用奥尔特-1177药理学抑制NLRP 3的机制，并确定NLRP 3阻断是否 抑制HSPC扩增、异常代谢活性和炎性免疫细胞在 心脏和其他组织。这项工作有可能重新框架CH作为一个治疗靶向代谢 表型，显著改善CH高患病率群体（如老年人）的健康。