Mechanisms and Modulation of Accelerated Atherosclerosis in Clonal Hematopoiesis

克隆造血加速动脉粥样硬化的机制和调节

• 批准号: 10590675
• 负责人: Peter Libby
• 金额: $ 65.04万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590675
• 关键词: Ablation; Acceleration; Age; Aging; Antibodies; Arterial Fatty Streak; Atherosclerosis; Biology; Blood; Bone Marrow; Bone Marrow Cells; Bone Marrow Stem Cell; Cardiovascular system; Cells; Chimera organism; Chimeric Proteins; Clinical; Clinical Data; DNMT3a; Data; Endothelium; Etiology; Event; Evolution; Female; Genotype; Gonadal Steroid Hormones; Hematopoiesis; Host Defense; Human; IL-6 inhibitor; IL6ST gene; IgG1; In Situ; Individual; Infection; Inflammasome; Inflammation; Inflammatory; Interleukin 6 Receptor; Interleukin-1; Interleukin-1 beta; Interleukin-6; Interleukins; Interruption; Lesion; Leukocytes; LoxP-flanked allele; Macrophage; Mediator; Messenger RNA; Monitor; Mononuclear; Mus; Mutation; Myelogenous; Myeloid Cells; Oncology; Organ; Outcome; PTPRC gene; Pathway interactions; Phagocytes; Population; Procedures; Proteins; Recurrence; Research; Risk; Risk Factors; Role; Science; Sex Differences; Signal Transduction; Smooth Muscle Myocytes; Somatic Mutation; Spleen; Testing; Work; atherogenesis; atherothrombosis; cardiovascular risk factor; clinical investigation; cytokine; experience; experimental study; genetic variant; improved; infection risk; interleukin-6 receptor alpha; male; mutant; novel; peripheral blood; preservation; response; single-cell RNA sequencing; targeted treatment; translational potential

Project Summary/Abstract With age, humans can accumulate leukocyte clones in blood that arise from somatic mutations in bone marrow stem cells that enhance expansion: clonal hematopoiesis of indeterminate potential (CHIP). Mutations in DNMT3A and TET2 account for the plurality of these clones. CHIP confers highly elevated cardiovascular (CV) risk, independent of traditional risk factors. We have found accelerated atherogenesis and the involvement of IL-1 and IL-6 in mice with myeloid loss of Tet2 or Dnmt3a function and that genetically reduced IL-6 signaling abrogates the elevated CV risk in humans with DNMT3A or TET2 CHIP. Analyses of our CANTOS trial showed greater efficacy of IL-1β inhibition in humans with DNMT3A or TET2 CHIP. These results point the way to a genotype-directed allocation of therapy, an approach that has transformed oncology but remains aspirational in atherosclerosis. The roles of classical vs. trans IL-6 signaling in atherothrombosis requires further study due to conflicting evidence. Specific aim 1 will test the hypothesis that atherosclerotic mice that mimic CHIP due to myeloid deficiency in Dnmt3a treated with an antibody that interrupts global IL-6 signaling by neutralizing IL-6 receptor α (IL-6r, CD126) have decreased atherosclerosis and inflammation within the lesions, blood and other organs as gauged in part by single-cell RNA sequencing (scRNA-seq). We further hypothesize that IL-6r inhibition will limit expansion of the mutant clone. Specific aim 2 will probe the role of classical vs. trans IL-6 signaling in myeloid cells in accelerated atherogenesis in CHIP using Il6rflox/flox/Lyz2-Cre mice bone marrow chimeric LDLR-/- mice (to block leukocyte classical signaling), and administration of a gp130-IgG1-Fc chimeric protein (to block trans signaling) using similar procedures and endpoints. The results will illuminate the controversy and unsettled science regarding the contributions of classical and trans IL-6 signaling to atherogenesis. Specific aim 3. Our preliminary experiments show that female myeloidTet2-/-Ldlr-/- mice have greater acceleration of atherogenesis than Tet2+/+ Ldlr-/- male mice and, unlike their male counterparts, show reduced atherogenesis with IL-1β neutralization. We will localize where this sex difference operates in the inflammasome–IL-1β–IL-6 pathway, and test the hypothesis that female Dnmt3a-/-Ldlr-/- mice have greater response to IL-6r inhibition than males. We will probe mechanisms by gonadal ablation experiments and by analysis of scRNA-seq data, which in preliminary data shows IL-1β expression in resident macrophages cells from atheroma from Tet2-/- females but not males. This work will deepen understanding of the mechanisms of accelerated atherosclerosis in CHIP. Our new pilot clinical data show that an anti-IL-6 antibody can mute inflammation in humans. Thus, the work proposed here will provide an indispensable step toward validating and furnishing the fundamental basis of an immediately translatable targeted therapeutic strategy based on CHIP genotype.

项目摘要/摘要 随着年龄的增长，人类可以在血液中积累血液中的白细胞克隆 增强膨胀的干细胞：不确定电势的克隆造血（CHIP）。突变 DNMT3A和TET2解释了这些克隆的多数。筹码供应高度升高的心血管（CV） 风险，独立于传统风险因素。我们发现加速了动脉粥样硬化和参与 具有TET2或DNMT3A功能损失的小鼠中的IL-1和IL-6，该基因降低了IL-6信号传导 用DNMT3A或TET2芯片消除了人类的CV风险升高。我们的Cantos试验的分析显示 DNMT3A或TET2芯片的人类IL-1β抑制的效率更高。这些结果指向了 基因型定向治疗的分配，这种方法已经改变了肿瘤学但在 动脉粥样硬化。经典与反式IL-6信号在动脉粥样硬化中的作用需要进一步研究 矛盾的证据。具体目标1将检验以下假设：动脉粥样硬化小鼠模仿芯片 用抗体处理的DNMT3A中的髓样缺乏，该抗体通过中和IL-6来中和全局IL-6信号传导 受体α（IL-6R，CD126）在病变，血液和其他病变内降低了动脉粥样硬化和炎症 器官作为单细胞RNA测序（SCRNA-SEQ）的一部分。我们进一步假设IL-6R 抑制作用将限制突变克隆的扩展。特定的目标2将探测经典与反式IL-6的作用 使用IL6rflox/flox/lyz2-cre小鼠骨髓在芯片中加速动脉粥样硬化的髓样细胞中的信号传导 嵌合LDLR - / - 小鼠（阻止白细胞经典信号传导），并给予GP130-IGG1-FC嵌合 蛋白质（以阻止反式信号传导）使用相似的过程和终点。结果将照亮 关于经典和反式IL-6信号对对 动脉粥样硬化。特定目标3。我们的初步实验表明雌性MyeloidTet2 - / - ldlr - / - 鼠标具有 与TET2+/+ LDLR - / - 雄性小鼠相比，动脉粥样硬化的加速度更大，并且与男性同行不同 通过IL-1β神经元化减少了动脉粥样硬化。我们将本地化这种性别差异在哪里运作 炎性体– IL-1β– IL-6途径，并测试雌性DNMT3A - / - LDLR - / - 小鼠的假设更大 对IL-6R抑制的反应比男性的抑制作用。我们将通过性腺消融实验探测机制，并通过 SCRNA-seq数据的分析，在初步数据中显示了居民巨噬细胞中IL-1β的表达 来自TET2 - / - 女性的动脉瘤，但不是男性。这项工作将加深对机制的理解 芯片中加速了动脉粥样硬化。我们的新试点临床数据表明，抗IL-6抗体可以静音 人类炎症。这是在这里提出的工作将为验证提供必不可少的步骤 并提供基于立即翻译的有针对性理论策略的基本基础 芯片基因型。