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 该提案的目的是确定靶向克隆造血的新方法（CH）。 CH是 以选择性扩展造血茎和祖细胞（HSPC）克隆的特征 TET2和DNMT3A等基因中的突变。这些HSPC又产生致病性突变体髓样细胞 这可能导致几种与衰老有关的合并症，包括心血管疾病（CVD）和全因 死亡。扩展的突变体HSPC池也可能导致血液系统恶性肿瘤的风险增加。 古老的人中，CH患病率显着升高，患有先前的遗传毒性暴露，吸烟的人 病史和/或慢性炎症性疾病。这些条件与长期扰动有关 生理稳态，其特征是过度炎症。了解促进的机制 突变HSPC的选择性扩展对于确定可以抑制CH的治疗靶标至关重要。 我们应用的主要前提是CH是由炎症之间的可靶向相互作用引起的 信号和更改的代谢编程，以支持充满活力的需求，从而获得首选 CH HSPC的扩展。使用鼠标作为代表人类关键特征的模型，我们的初步 数据表明，CH HSPC暴露于转录因子HIF-1异常糖酵解的水平增加 相对于野生型HSPC，代谢和ATP产生增加。我们发现炎性细胞因子， 特别是IL-1强烈增强CH HSPC的扩张，糖酵解代谢和HIF-1活性。令人惊讶的是， 用OLT-1177（一种NLRP3炎性体抑制剂）处理CH小鼠，可防止裂解和激活 IL-1的可能抑制CH。我们假设CH是相互依赖机制的结果 NLRP3介导的IL-1生产电位HIF-1和下游糖酵解活性，以支持CH HSPC 扩张。我们建议NLRP3抑制作用破坏了该电路，从而限制了CH HSPC的扩展。 为了解决该机制，我们提出了两个具体目的：1）我们将表征 TET2/，TET2+/和DNMT3AR878H/+ CH HSPC使用基于体内的质谱和基于流式的细胞衡 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 和糖酵解的能量需求； 2）我们将评估NLRP3的要求 CH HSPC的糖酵解活性和/或优选膨胀。我们将使用分子遗传学方法 评估HIF-1和NLRP3在调节CH HSPC代谢中的作用和机制并促进其 体内扩展。使用CH的非调节自适应BM传输鼠标模型，我们将验证我们的 使用OLT-1177对NLRP3的药物抑制的机制，并确定NLRP3 blocade是否 抑制HSPC的扩张，异常代谢活性和炎性免疫细胞的积累 心脏和其他组织。这项工作有可能重新构架CH作为一种治疗靶向代谢 表型，可以显着改善CH较高患病率的组之间的健康状况。