Mechanisms of Atherogenesis in Clonal Hematopoiesis

克隆造血中动脉粥样硬化形成的机制

• 批准号: 10852373
• 负责人: Trevor Perawaskin Laramee Fidler
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 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 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10852373
• 关键词: 8-hydroxy-2' -deoxyguanosine; Acceleration; Age; Antibodies; Area; Arterial Fatty Streak; Arteries; Atherosclerosis; Awareness; Bar Codes; Bioinformatics; Blood Cells; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; CASP1 gene; Cancer Patient; Cardiovascular Diseases; Caspase; Cell Death; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Characteristics; Clonal Expansion; Collaborations; Complement; Coronary; Coronary heart disease; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Gene; DNA biosynthesis; DNA lesion; Development; Elderly; Endowment; Environment; Epitopes; Etiology; Genes; Goals; Heart Diseases; Hematopoiesis; Hematopoietic; Histology; Human; Immune; Inflammasome; Inflammation; Inflammatory; Interleukin-1; Interleukin-1 Receptors; Interleukin-1 beta; Knock-out; Knowledge; Lead; Learning; Lesion; Macrophage; Mentors; Metabolism; Minority; Modeling; Mus; Mutate; Mutation; Myelogenous; Myeloid Cells; Myocardial Infarction; Necrosis; PTPRC gene; Pathway interactions; Patients; Phase; Phenotype; Population; Proliferating; Protein phosphatase; Reactive Oxygen Species; Reporting; Research; Research Personnel; Resolution; Risk; Risk Factors; Role; Signal Transduction; Smooth Muscle Myocytes; Somatic Mutation; Source; Stroke; TP53 gene; Technology; Testing; Thinness; Training; Transcript; Transgenic Organisms; Transplantation; Variant; Vascular Smooth Muscle; atherogenesis; blood fractionation; cancer survival; cardiovascular disorder risk; chemotherapy; ds-DNA; extracellular; gain of function; genetic variant; improved; indexing; mouse model; mutant; novel; oxidation; oxidative DNA damage; premature; prevent; programs; replication stress; response; single-cell RNA sequencing; skills; targeted treatment; transcriptome; transcriptome sequencing

1 Clonal hematopoiesis (CH), a highly prevalent condition in the elderly, arises from somatic mutations that endow 2 a proliferative advantage to a subset of hematopoietic cells. CH increases the risk of myocardial infarction and stroke 3 independent of traditional risk factors. Relative to other common genetic variants giving rise to CH, the JAK2V617F (JAK2VF) 4 mutation that increases JAK/STAT signaling occurs at a younger age and imparts a substantial risk of premature coronary 5 heart disease (CHD). In CH mutated cells make up only a small fraction of blood cells, however, patients maintain an 6 elevated risk of CHD, suggesting mutant cells may have an outsized impact on their environment. Using murine models of 7 Jak2VF CH we found that IL-1 specifically promotes cell intrinsic proliferation of Jak2VF macrophages, but not wild type 8 (WT) macrophages even within the same atherosclerotic lesions. Administration of antibodies to IL-1 reduced Jak2VF 9 macrophage proliferation and improved features of plaque stability. scRNA-Seq analysis of lesion from mice with Jak2VF 10 expression in all bone marrow cells identified a prominent inflammatory myeloid population that was enriched for genes 11 associated with IL-1 signaling. We found that inflammatory myeloid cell abundance was decreased in the absence of the 12 pyroptosis executioner Gasdermin D (Gsdmd), potentially due to suppressed IL-1 secretion. Therefore, Specific Aim 1 13 will investigate the ability of Jak2VF cells to promote inflammatory phenotypes in WT cells within the same lesion through 14 IL-1 signaling. Mechanistically we have shown that the AIM2 inflammasome is activated in lesions from Jak2VF mice. 15 Deletion of Gsdmd from Jak2VF cells leads to increased DNA double strand breaks in lesions. Bone marrow derived 16 macrophages expressing Jak2VF, but lacking Caspase 1/11 have increased reactive oxygen species, suggesting DNA damage 17 occur upstream of inflammasome activation. Thus, Specific Aim 2 will determine if increased oxidized DNA in Jak2VF 18 macrophages promotes inflammasome activation and inflammation in lesions. Specific Aim 3 will then investigate if 19 truncation mutations in the CH variant protein phosphatase Mg2+/Mn2+ dependent 1D (PPM1D) that leads to inactivation of 20 DNA repair proteins, promotes atherosclerosis. The research proposed here will be accomplished by expanding my 21 conceptual knowledge of DNA damage in macrophages and atherosclerosis and will be complemented by training in single 22 cell sequencing technology to understand how immune cell populations change in lesions based on CH variant status. This 23 training will be supported by rigorous mentoring and collaboration with my scientific advisors during the K99 phase. 24 Together the proposed research and training will elucidate mechanism by which DNA damage and inflammasome activation 25 drive accelerated atherosclerosis in CH which will provide me the knowledge and skills to successfully transition into an 26 independent academic researcher. 27 28 29 30

1克隆性造血（CH），一种在老年人中非常普遍的疾病，由体细胞突变引起， 2对造血细胞亚群具有增殖优势。CH增加心肌梗死和中风的风险 3独立于传统的危险因素。相对于引起CH的其他常见遗传变异，JAK 2 V617 F（JAK 2 VF） 增加JAK/STAT信号传导的4突变发生在较年轻的年龄，并赋予过早的冠状动脉粥样硬化的显著风险。 5心脏病（CHD）。在CH中，突变的细胞仅占血细胞的一小部分，然而，患者保持了 6冠心病的风险增加，这表明突变细胞可能对其环境产生巨大影响。使用小鼠模型， 7 Jak 2 VF CH我们发现IL-1 β特异性促进Jak 2 VF巨噬细胞的细胞内源性增殖，而不是野生型 8（WT）巨噬细胞，即使在相同的动脉粥样硬化病变。施用针对IL-1的抗体可减少Jak 2 VF 9巨噬细胞增殖和改善斑块稳定性的特征。来自具有Jak 2 VF的小鼠的病变的scRNA-Seq分析 10在所有骨髓细胞中的表达鉴定了一个突出的炎性骨髓细胞群， 11与IL-1信号相关。我们发现，在缺乏免疫抑制剂的情况下， 12焦亡执行者Gasdermin D（Gsdmd），可能是由于抑制IL-1 β分泌所致。具体目标1 13将研究Jak 2 VF细胞促进相同病变内WT细胞中炎性表型的能力， 14 IL-1信号转导。从机制上讲，我们已经表明，AIM 2炎性小体在来自Jak 2 VF小鼠的病变中被激活。 15 Jak 2 VF细胞中Gsdmd的缺失导致病变中DNA双链断裂增加。骨髓来源 16例表达Jak 2 VF但缺乏Caspase 1/11的巨噬细胞活性氧增加，提示DNA损伤 17个发生在炎性小体激活的上游。因此，特定目标2将确定Jak 2 VF中的氧化DNA是否增加 18巨噬细胞促进损伤中的炎性体活化和炎症。具体目标3将调查，如果 CH变体蛋白磷酸酶Mg 2 +/Mn 2+依赖性1D（PPM 1D）中的19个截短突变导致 20 DNA修复蛋白，促进动脉粥样硬化。这里提出的研究将通过扩展我的 21巨噬细胞和动脉粥样硬化中DNA损伤的概念性知识，并将通过单 22细胞测序技术，以了解免疫细胞群体如何根据CH变异状态在病变中变化。这 在K99阶段，培训将得到严格的指导和我的科学顾问的合作。 24拟议的研究和培训将共同阐明DNA损伤和炎性体激活的机制 25驱动加速动脉粥样硬化在CH这将为我提供的知识和技能，成功地过渡到一个 26篇独立学术研究 27 28 29 30