Project 2: Cholesterol Regulation of Inflammatory Macrophages in Atherosclerosis

项目2：动脉粥样硬化中炎症巨噬细胞的胆固醇调节

• 批准号: 10334095
• 负责人: Yury Miller
• 金额: $ 4.3万
• 依托单位: LA JOLLA INSTITUTE FOR IMMUNOLOGY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-06-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10334095
• 关键词: ATAC-seq; ATP binding cassette transporter 1; Agonist; Animal Model; Antibodies; Apolipoprotein A-I; Atherosclerosis; Automobile Driving; B-Lymphocyte Subsets; B-Lymphocytes; Binding Proteins; Biological Assay; Cardiovascular Diseases; Cell Communication; Cell physiology; Cells; Cellular Membrane; ChIP-seq; Cholesterol; Cholesterol Homeostasis; Chronic; Clinical; Complex; Cultured Cells; Cyclodextrins; DNA Sequencing Facility; Data; Desmosterol; Development; Diagnostic; Dimerization; Epigenetic Process; Epitopes; Excision; Experimental Designs; Feedback; Flow Cytometry; Gatekeeping; Gene Expression; Genetic Transcription; Human; Image; Immune; Immunity; Inflammation; Inflammatory; Inflammatory Response; Intervention; Knock-out; Knockout Mice; Leukocytes; Ligands; Liquid substance; Longitudinal cohort; Macromolecular Complexes; Malondialdehyde; Mediating; Membrane; Membrane Microdomains; Metabolic; Morbidity - disease rate; Mus; Myocardial Infarction; Oxides; Patients; Peptide Sequence Determination; Peripheral Blood Mononuclear Cell; Phenotype; Phospholipids; Play; Proteins; Regulation; Resources; Role; Signal Transduction; Solid; Source; Stroke; T-Lymphocyte; TLR4 gene; Testing; Training; Transgenic Mice; beta-Cyclodextrins; cholesterol control; diagnostic value; epigenetic memory; hypercholesterolemia; macrophage; metabolomics; mevalonate; mimetics; monocyte; mortality; novel strategies; overexpression; oxidation; oxidized lipid; peripheral blood; prevent; prognostic; prognostic value; receptor; recruit; scaffold; single cell proteins; transcriptome sequencing

Project 2 Summary Atherosclerosis is the primary cause of cardiovascular disease (CVD), which manifests in myocardial infarction and stroke, a major source of mortality and morbidity in the US. Hypercholesterolemia and chronic inflammation are leading causative factors in the development of atherosclerosis. Many inflammatory receptors and other proteins involved in the inflammatory response localize and become activated in cholesterol-rich membrane microdomains, often designated as lipid rafts, which provide a solid platform for protein assembly within a liquid membrane. Here, we propose the concept of inflammarafts, enlarged lipid rafts hosting assembled inflammatory signaling complexes, to help formalize the lipid raft-centric view of inflammation and its control by cholesterol metabolism. Cholesterol depletion from inflammarafts in activated cells disrupts inflammatory signaling, and the approaches that allow for targeting cholesterol removal selectively to inflammatory cells may be used for atheroprotection. In the context of atherosclerosis, monocytes and macrophages develop long-term adaptation, or trained immunity, which results in sustained inflammatory phenotypes and includes epigenetic memory, as well as transcriptional and metabolic alterations leading to a chronic inflammatory state. In this application, we will test the hypotheses that inflammarafts serve as a signaling platform that mediates cell reprogramming and that trained immune cells sustain lipid rafts to maintain the inflammatory response. We propose that this positive feedback mechanism between inflammarafts and metabolic, gene expression and epigenetic alterations plays a major role in the development of atherosclerosis. Specifically, to test the hypothesis that inflammarafts serve as gatekeepers and effectors of trained immunity in atherosclerosis, we will use inflammaraft assays to image and quantify lipid rafts, accessible cholesterol, membrane order, TLR4 dimerization and the assembly of other receptor complexes. RNA-seq, ATAC-seq, ChIPseq and metabolomics will be used to characterize trained immunity. To modulate ligand-dependent inflammaraft assembly, we will neutralize oxidized lipid DAMPs, abundant in plaques, in transgenic mice that overexpress oxidation-specific antibodies. APOA1, AAV-AIBP (apoA-I binding protein) and cyclodextrin interventions will be used for systemic and targeted depletion of cholesterol. Conversely, cholesterol and inflammarafts will be increased in inducible, macrophage-specific ABCA1/ABCG1 knockout mice. Statins/mevalonate and desmosterol mimetics will be used to modulate macrophage reprogramming. Importantly, we will evaluate the diagnostic and prognostic power of inflammaraft phenotyping of blood leukocytes in CVD. By using the resources of PPG’s clinical and single cell protein and RNA sequencing cores, we will test the hypotheses that the abundance of inflammarafts and raft-dependent receptor assemblies in peripheral blood monocytes and T and B cells reflect their trained immunity and inflammatory gene expression phenotypes and can be used for diagnostic and prognostic purposes in patients with CVD.

项目2摘要 动脉粥样硬化是心血管疾病（CVD）的主要原因，它在心肌梗塞中表现出来 和中风，这是美国死亡率和发病率的主要来源。高胆固醇血症和慢性血症 炎症是动脉粥样硬化发展的主要因素。许多炎症受体 和其他参与炎症反应的蛋白质定位并在富含胆固醇中激活 膜微区通常被指定为脂质筏，为蛋白质组装提供了坚实的平台 在液体膜中。在这里，我们提出了炎症的概念，放大的脂筏托管 组装炎症信号传导复合物，以帮助形式上以脂质筏为中心的炎症和 胆固醇代谢的控制。活化细胞中炎症的胆固醇耗竭 炎症信号传导以及允许靶向胆固醇选择性去除的方法 炎性细胞可用于动脉保护。在动脉粥样硬化，单核细胞和 巨噬细胞长期适应或受过训练的免疫培养物，导致持续的炎症 表型，包括表观遗传记忆，以及转录和代谢改变导致 慢性炎症状态。在此应用中，我们将检验炎性玛拉夫作为一个的假设 介导细胞重编程和训练的免疫细胞的信号平台维持脂质筏 保持炎症反应。我们建议这种积极的反馈机制 炎症和代谢，基因表达和表观遗传学改变在发育中起主要作用 动脉粥样硬化。具体而言，要检验炎症是守门人的假设和 在动脉粥样硬化中受过训练的免疫力，我们将使用炎症测定法来形象和量化脂质筏， 可访问的胆固醇，膜顺序，TLR4二聚化和其他受体复合物的组装。 RNA-SEQ，ATAC-SEQ，CHIPSEQ和代谢组学将用于表征训练有素的免疫功能。调制 配体依赖性炎症组件，我们将中和氧化的脂质潮湿，斑块丰富，在 过表达氧化特异性抗体的转基因小鼠。 ApoA1，AAV-AIBP（ApoA-I结合蛋白） 环糊精干预措施将用于胆固醇的全身和靶向耗竭。反过来， 在诱导的，巨噬细胞特异性的ABCA1/ABCG1敲除中，胆固醇和炎症将增加 老鼠。他汀类药物/甲氯苯二甲酸盐和脱乳酚模拟物将用于调节巨噬细胞重编程。 重要的是，我们将评估血液炎症表型的诊断和预后能力 CVD中的白细胞。通过使用PPG的临床和单细胞蛋白和RNA测序核的资源， 我们将测试假设炎症和筏依赖受体组件的抽象 外周血单核细胞和T和B细胞反映其受过训练的免疫抑制和炎症基因表达 表型，可用于CVD患者的诊断和预后目的。