Novel pathways controlling macrophage inflammation and resolution in atherosclerosis

控制巨噬细胞炎症和动脉粥样硬化消退的新途径

• 批准号: 10450684
• 负责人: GOKHAN S HOTAMISLIGIL
• 金额: $ 72.1万
• 依托单位: HARVARD SCHOOL OF PUBLIC HEALTH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10450684
• 关键词: Anti-Inflammatory Agents; Atherosclerosis; Blood Vessels; Bone Marrow; Cardiac; Cardiovascular Diseases; Cell model; Cells; Cholesterol; Cholesterol Homeostasis; Chronic; Clinical Trials; Coupled; DNA sequencing; Data; Dependovirus; Disease; Genes; Genomics; Goals; Hepatocyte; Immune; Immunoprecipitation; Immunosuppression; Impairment; Inflammation; Inflammatory; Intervention; Lesion; Light; Link; Lipids; Low-Density Lipoproteins; Mediating; Mediator of activation protein; Metabolism; Methodology; Mission; Modeling; Molecular; Molecular Mechanisms of Action; Mus; Outcome; Output; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Preparations; Phase; Play; Post-Translational Protein Processing; Process; Progressive Disease; Proteins; Public Health; Publishing; Regulation; Research; Research Proposals; Resolution; Role; Stimulus; System; Testing; Therapeutic; Tissues; United States National Institutes of Health; Work; atherogenesis; atheroprotective; base; gain of function; global health; in vivo; innovation; insight; lipid metabolism; loss of function; macrophage; mortality; mouse model; new therapeutic target; novel; novel therapeutic intervention; nuclear factor-erythroid 2; overexpression; prevent; response; therapeutic target; tissue repair; transcription factor; western diet

Summary In-spite of effective strategies for lowering cholesterol, atherosclerosis and associated cardiovascular diseases remain a major global health burden. Work in recent years has highlighted the exciting therapeutic potential for interventions that reduce inflammation, as well as the need for identifying novel and selective targets. Our long term goal is to understand how lipid metabolism and inflammation interact, and how these interactions can be useful therapeutically. The objective of the current research proposal is to determine the role of the transcription factor Nuclear factor erythroid 2 related factor-1 (Nrf1; also known as Nfe2L1) in linking these two processes. The central hypothesis is that Nrf1 in the macrophage is a critical factor for preventing chronic inflammation and maintaining cholesterol homeostasis, and that enhancing these actions of Nrf1 may be beneficial in atherosclerosis. This is based on compelling new preliminary data showing that Nrf1 is activated in macrophages in response to inflammatory stimuli and is necessary for the expression of lipid metabolism pathway genes that are crucial for proper resolution of the inflammatory state. Previous results also showed that deletion of Nrf1 drastically impairs cellular cholesterol homeostasis.The rationale for the proposed research is that understanding the function and mechanism of macrophage Nrf1 may provide important insight into regulation of macrophage inflammation and resolution in general and reveal a novel therapeutic target for the treatment of atherosclerosis. The central hypothesis will be tested by pursuing three specific aims: 1) Examine the function of Nrf1 in macrophages; 2) Elucidate the molecular mechanism of Nrf1 action in macrophages; and 3) Explore the importance of macrophage-Nrf1 in atherosclerosis in-vivo. Under the first aim, tissue-specific inducible deletion models established and tested in the applicant's lab will be used for a thorough characterization of the function of Nrf1 in inflammatory output, lipid metabolism and resolution of inflammation. This aim will also explore the impact of cholesterol on Nrf1 function, and the reciprocal role of Nrf1 in regulating cholesterol homeostasis in macrophages. In the second aim, immunoprecipitation of readily available tagged protein coupled to DNA sequencing and MS/MS analysis will identify, protein interactors and post translational modifications of Nrf1 potentially mediating its function. The third aim will test the impact of Nrf1 deficiency and overexpression on atherosclerosis in-vivo using already established Western-diet fed Ldlr-deficient mice with either macrophage specific deletion of Nrf1, or adeno-associated-virus (AAV) mediated macrophage specific overexpression of Nrf1. The approach is innovative because it attempts to overcome issues of pro-resolving mediator delivery by using naturally existing systems within macrophages. The proposed research is significant, because current approaches to activate endogenous resolution of inflammation in the context of atherosclerosis are limited, and this study might provide important insight into the role of the resolution in atherosclerosis and reveal a novel therapeutic target.

概括 尽管有降低胆固醇、动脉粥样硬化和相关心血管疾病的有效策略 仍然是全球主要的健康负担。近年来的工作凸显了令人兴奋的治疗潜力 减少炎症的干预措施，以及确定新的和选择性目标的需要。我们的长 术语目标是了解脂质代谢和炎症如何相互作用，以及这些相互作用如何 有治疗作用。当前研究计划的目标是确定转录的作用 连接这两个过程的核因子红细胞 2 相关因子 1（Nrf1；也称为 Nfe2L1）。 核心假设是巨噬细胞中的 Nrf1 是预防慢性炎症和 维持胆固醇稳态，增强 Nrf1 的这些作用可能有益于 动脉粥样硬化。这是基于令人信服的新初步数据，表明 Nrf1 在巨噬细胞中被激活 响应炎症刺激并且对于脂质代谢途径基因的表达是必需的 对于正确解决炎症状态至关重要。先前的结果还表明，Nrf1 的缺失 严重损害细胞胆固醇稳态。拟议研究的基本原理是了解 巨噬细胞Nrf1的功能和机制可能为巨噬细胞的调控提供重要的见解 炎症和消退的总体情况，并揭示了治疗动脉粥样硬化的新治疗靶点。 将通过追求三个具体目标来检验中心假设：1）检查 Nrf1 在 巨噬细胞； 2）阐明Nrf1在巨噬细胞中作用的分子机制； 3) 探索 巨噬细胞-Nrf1 在体内动脉粥样硬化中的重要性。第一个目标是组织特异性诱导缺失 在申请人实验室建立和测试的模型将用于全面表征功能 Nrf1 在炎症输出、脂质代谢和炎症消退中的作用。这一目标还将探索 胆固醇对 Nrf1 功能的影响，以及 Nrf1 在调节胆固醇稳态中的相互作用 巨噬细胞。第二个目标是对现成的 DNA 标记蛋白进行免疫沉淀 测序和 MS/MS 分析将鉴定 Nrf1 的蛋白质相互作用因子和翻译后修饰 潜在地调节其功能。第三个目标将测试 Nrf1 缺乏和过度表达对 使用已经建立的西方饮食喂养的 Ldlr 缺陷小鼠和巨噬细胞进行体内动脉粥样硬化实验 Nrf1 的特异性缺失，或腺相关病毒 (AAV) 介导的 Nrf1 巨噬细胞特异性过表达。 该方法具有创新性，因为它试图通过使用 巨噬细胞内自然存在的系统。拟议的研究意义重大，因为目前 在动脉粥样硬化背景下激活炎症内源性消退的方法是有限的，并且 这项研究可能为了解该解决方案在动脉粥样硬化中的作用提供重要见解，并揭示一种新的机制 治疗目标。