Mechanisms of Atherogenesis in Clonal Hematopoiesis

克隆造血中动脉粥样硬化形成的机制

基本信息

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
项目类别：
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&apos -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

项目摘要

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 CH 造血细胞亚群的增殖优势会增加心肌梗塞和中风的风险。 3 相对于导致 CH 的其他常见遗传变异，JAK2V617F (JAK2VF) 独立。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) 中，突变细胞仅占血细胞的一小部分，但患者仍保持着 6 冠心病风险增加，表明突变细胞可能对其环境产生巨大影响。 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，即使在相同的动脉粥样硬化病变内，给予 IL-1 抗体也会减少 Jak2VF。 9 对 Jak2VF 小鼠病变进行 scRNA-Seq 分析，发现巨噬细胞增殖和斑块稳定性得到改善。 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 IL-1 信号的情况下，炎症骨髓细胞丰度会降低。 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 信号传导从机制上来说，我们已经证明 AIM2 炎症小体在 Jak2VF 小鼠的病变中被激活。 15 Jak2VF 细胞中 Gsdmd 的缺失会导致骨髓损伤中 DNA 双链断裂的增加。 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 发生在炎性体激活的上游，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炎症病变。具体目标 3 将研究是否 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研究来完成。 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。 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