Complement Protein C1q in Atherosclerosis

动脉粥样硬化中的补体蛋白 C1q

基本信息

批准号：
10117075
负责人：
Deborah Ann Fraser
金额：
$ 11.06万
依托单位：
CALIFORNIA STATE UNIVERSITY LONG BEACH
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-05 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10117075
关键词：
25-hydroxycholesterol Acute Adhesions Apoptosis Apoptotic Arterial Fatty Streak Arterial Intimas Atherosclerosis Autoimmune Diseases Binding Biological Biological Assay Blood Vessels Cardiovascular Diseases Cause of Death Cell Adhesion Molecules Cell Survival Cell physiology Cells Chemotaxis Cholesterol Chronic Classical Complement Pathway Clinical Complement Complement 1q Complement Activation Cytokine Gene Data Disease Disease Progression Endothelial Cells Endothelium Environment Enzymes Excision Foam Cells Gene Expression Goals Harvest Homeostasis Human Hyperlipidemia Immune Immune response In Vitro Inflammation Inflammatory Innate Immune System Investigation Ischemic Stroke Lesion Lipids Lipoproteins Liver X Receptor Low-Density Lipoproteins Mass Spectrum Analysis Measures Mixed Function Oxygenases Molecular Molecular Mechanisms of Action Mus Myocardial Infarction Necrosis Normal tissue morphology Nuclear Outcome Pathology Pathway interactions Pattern recognition receptor Peptide Hydrolases Perception Permeability Phagocytes Phagocytosis Plasma Play Production Proteins Public Health Reactive Oxygen Species Receptor Activation Reporting Research Role Rupture Signal Transduction Subendothelial Layer Testing Thrombus Vascular Diseases Vascular Endothelial Cell Vascular Endothelium Woman chemokine complement pathway cytokine immunoregulation improved in vitro testing in vivo in vivo Model lipid metabolism lipidomics macrophage men migration monocyte mouse model new therapeutic target novel pathogen programs response therapeutic target

项目摘要

PROJECT SUMMARY Complement Protein C1q in Atherosclerosis Atherosclerosis is a chronic inflammatory disorder which in early stages is characterized by the migration of modified lipoproteins and macrophages into the arterial intima and the formation and apoptosis of macrophage foam cells. In late stages of disease, inadequate/defective apoptotic foam cell removal by macrophages leads to their secondary necrosis and plaque formation. Damage to this plaque by pro-inflammatory cytokines, proteases and oxygen radicals can cause rupture and thrombus formation, and acute clinical complications such as myocardial infarction and ischemic stroke. Thus, inflammation, macrophages and vascular integrity are key in progression of this disease. Activation of complement has been shown to contribute to inflammation and exacerbate pathology. However, studies in mice deficient in the first protein in the classical complement pathway, C1q, suggest that this protein actually has a protective role in the early atherosclerotic lesion. Innate immune protein C1q is not only able to trigger the inflammatory complement cascade, but is also a pattern recognition receptor that opsonizes targets and directly interacts with phagocytes and other cells. Interaction activates responses including phagocytosis of targets such as apoptotic cells or damaged molecules, and modulation of cytokine and gene expression. Therefore, our central hypothesis is that complement-independent actions of C1q program protective, anti-atherosclerotic cellular responses in atherosclerosis. Our recent studies have demonstrated C1q modulation of macrophage inflammatory polarization in vitro and in vivo models of atherosclerosis and identified several molecular mechanisms involved. These studies provided preliminary data that C1q modulates vascular endothelial responses in atherosclerosis. In addition, data suggest that C1q increases levels of bioactive oxysterols 24- and 25- hydroxycholesterol by macrophages in response to hyperlipidemic conditions in vitro and in vivo. The goal of this project is to broaden our understanding of C1q molecular interactions beyond macrophages to the entire lesional environment. Specific aims are: 1: Investigate C1q modulation of vascular responses in atherosclerosis. We will test the hypothesis that C1q reduces monocyte chemotaxis and improves vascular integrity in atherosclerosis. This will be tested using primary human monocytes and vascular endothelial cells to perform chemotaxis, transendothelial migration and permeability assays. In addition, a comprehensive exploration of chemokines and adhesion molecules produced by macrophages and vascular endothelial cells in response to modified LDL in the presence or absence of C1q will be performed using human cells and in plasma harvested from C1q sufficient and deficient hyperlipidemic mice. Specific Aim 2: Investigate C1q modulation of lipid metabolism in atherosclerosis. These studies will test the hypothesis that C1q modulation of oxysterols is involved in macrophage foam cell survival and polarization. Oxysterol levels will be measured in vitro, in primary human macrophages, and in vivo in plasma from a mouse model of hyperlipidemia, using mass spectrometry. Survival and polarization assays will be performed in specific pathway-deficient macrophages to identify their relative importance in these biological responses. Overall, these studies aim to explore the dual role that C1q plays in atherosclerosis, and should assist in identifying novel molecular pathways for therapeutic targeting in this disease.

项目摘要在动脉粥样硬化中补充蛋白C1q 动脉粥样硬化是一种慢性炎症性疾病，在早期，其特征是修饰的迁移脂蛋白和巨噬细胞进入动脉内膜以及巨噬细胞泡沫细胞的形成和凋亡。晚了疾病阶段，巨噬细胞去除凋亡泡沫细胞不足/缺陷导致其继发性坏死和斑块形成。促炎性细胞因子，蛋白酶和氧自由基对该斑块的损害可能导致破裂和血栓形成以及急性临床并发症，例如心肌梗塞和缺血性中风。因此，炎症，巨噬细胞和血管完整性是该疾病进展的关键。补体的激活被证明会导致炎症和加剧病理学。但是，对缺乏第一种蛋白质的小鼠的研究在经典的补体途径中，C1Q表明该蛋白在早期实际上具有保护作用动脉粥样硬化病变。先天免疫蛋白C1q不仅能够触发炎症补体级联反应，而且是还有一个模式识别受体，可以使靶标打击并直接与吞噬细胞和其他细胞相互作用。相互作用激活反应，包括吞噬细胞或凋亡细胞或受损分子的吞噬作用，并调节细胞因子和基因表达。因此，我们的核心假设是C1Q程序的无关行为动脉粥样硬化中的保护性，抗动脉粥样硬化的细胞反应。我们最近的研究表明C1Q 巨噬细胞在体外和体内模型的巨噬细胞炎性极化的调节，并确定了几个涉及的分子机制。这些研究提供了C1Q调节血管内皮的初步数据动脉粥样硬化的反应。此外，数据表明C1Q增加了24和25-的生物活性氧甲醇水平。巨噬细胞响应于体外和体内的高脂症状的羟基胆固醇。目标的目标项目是为了扩大我们对巨噬细胞以外的C1Q分子相互作用的理解到整个病变环境。具体目的是：1：研究动脉粥样硬化中血管反应的C1Q调节。我们将测试 C1Q降低单核细胞趋化性并改善动脉粥样硬化中的血管完整性的假设。这将是使用原代人单核细胞和血管内皮细胞进行测试以进行趋化性，跨内皮迁移和渗透性测定。此外，对趋化因子和粘附分子的全面探索在存在或不存在C1Q的情况下，巨噬细胞和血管内皮细胞响应于修饰的LDL 使用人类细胞和从C1Q收集的血浆进行足够和缺乏的高脂血症小鼠进行。具体的目标2：研究动脉粥样硬化中脂质代谢的C1Q调节。这些研究将检验以下假设氧化酚的C1Q调节参与巨噬细胞泡沫细胞的存活和极化。氧化酚水平将是在体外测量的原代人巨噬细胞和使用小鼠高脂血症的小鼠模型的血浆中的体内测量质谱法。生存和极化测定将在特定途径缺陷的巨噬细胞中进行确定它们在这些生物学反应中的相对重要性。总体而言，这些研究旨在探索C1Q的双重作用在动脉粥样硬化中发挥作用，应有助于鉴定该疾病中治疗靶向的新型分子途径。