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: 10535483
• 负责人: Trevor Perawaskin Laramee Fidler
• 金额: $ 16.55万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-15 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10535483
• 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) 是一种在老年人中非常普遍的疾病，由体细胞突变引起，该突变赋予 2 造血细胞亚群的增殖优势。 CH 增加心肌梗死和中风的风险 3.独立于传统风险因素。相对于导致 CH 的其他常见遗传变异，JAK2V617F (JAK2VF) 增加 JAK/STAT 信号传导的 4 突变发生在较年轻的年龄，并会增加冠状动脉早发的风险 5 心脏病（CHD）。在 CH 中，突变细胞仅占血细胞的一小部分，然而，患者仍保持 6 患冠心病的风险增加，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 Jak2VF CH 我们发现 IL-1 特异性促进 Jak2VF 巨噬细胞的细胞内在增殖，但野生型则不然 8 (WT) 巨噬细胞甚至在同一动脉粥样硬化病变内。 IL-1抗体的施用减少了Jak2VF 9.巨噬细胞增殖和斑块稳定性改善的特点。对 Jak2VF 小鼠病变进行 scRNA-Seq 分析 10 在所有骨髓细胞中的表达确定了一个显着的炎症性骨髓细胞群，该细胞群富含基因 11 与 IL-1 信号传导相关。我们发现，在缺乏 12 焦亡刽子手 Gasdermin D (Gsdmd)，可能是由于 IL-1 分泌受到抑制。因此，具体目标 1 13 将研究 Jak2VF 细胞通过以下方式促进同一病变内 WT 细胞炎症表型的能力 14 IL-1 信号传导。从机制上讲，我们已经证明 AIM2 炎症小体在 Jak2VF 小鼠的病变中被激活。 15 从 Jak2VF 细胞中删除 Gsdmd 会导致病变部位 DNA 双链断裂增加。骨髓来源 16 个表达 Jak2VF 但缺乏 Caspase 1/11 的巨噬细胞活性氧含量增加，表明 DNA 损伤 17 发生在炎症小体激活的上游。因此，特定目标 2 将确定 Jak2VF 中氧化 DNA 是否增加 18 巨噬细胞促进炎症小体激活和病变炎症。具体目标 3 将调查是否 CH 变体蛋白磷酸酶 Mg2+/Mn2+ 依赖性 1D (PPM1D) 中的 19 个截短突变导致 20种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 DNA 损伤和动脉粥样硬化的概念知识，并将通过单一培训进行补充 22 细胞测序技术可根据 CH 变异状态了解病变中免疫细胞群的变化。这 23 培训将在 K99 阶段得到我的科学顾问的严格指导和合作的支持。 24 拟议的研究和培训将共同阐明 DNA 损伤和炎症小体激活的机制 25 在 CH 加速动脉粥样硬化，这将为我提供知识和技能，以成功过渡到 26名独立学术研究员。 27 28 29 30