Oxidant resistant apoA1 in reverse cholesterol transport, inflammation and atherosclerosis

抗氧化剂 apoA1 在逆转胆固醇转运、炎症和动脉粥样硬化中的作用

基本信息

批准号：
9276118
负责人：
Jonathan D Smith
金额：
$ 39.63万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-23 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9276118
关键词：
ATP binding cassette transporter 1 Acute-Phase Reaction Affinity Aftercare Anti-Inflammatory Agents Anti-inflammatory Apolipoproteins Area Arterial Fatty Streak Atherosclerosis Bone Marrow Breeding Cell Adhesion Molecules Cell Membrane Proteins Cell Proliferation Cells Cholesterol Collagen Complete Blood Count Coronary Arteriosclerosis Data Development Endothelial Cells Endotoxins Environment Enzymes Epitopes Event Excretory function Gene Chips Genetic screening method Goals High Density Lipoprotein Cholesterol High Density Lipoproteins Human Human Genetics Impairment Individual Inflammation Inflammatory Knowledge Lead Lesion Lipids Lipoproteins Liver Measures Mediating Modeling Modification Molecular Profiling Monoclonal Antibodies Monocytosis Mus Myeloid Cells NF-kappa B Names Necrosis Neutrophilia Oxidants Oxides Pathway interactions Peripheral Peroxidases Pharmaceutical Preparations Phenylalanine Phospholipids Plasma Play Protein Isoforms Proteins Randomized Recombinants Reporting Resistance Risk Role Sepsis Series Site-Directed Mutagenesis Smooth Muscle Actin Staining Method Stimulus Testing Therapeutic Tissues Transgenic Mice Triglycerides Tryptophan Variant Zymosan antimicrobial atheroprotective cardiovascular disorder risk comparative efficacy cytokine disorder risk epidemiology study genetic variant histological stains humanized monoclonal antibodies loss of function macrophage monocyte mouse model neutrophil novel overexpression oxidation particle prevent protective effect reverse cholesterol transport therapeutic target

项目摘要

Abstract High levels of high density lipoprotein-cholesterol (HDL-C) are associated with lowered risk for cardiovascular disease (CVD) in epidemiological studies. Although several mechanisms may play a role in HDL’s protective effect, HDL and its major protein constituent, apolipoprotein-AI (apoA1), are critical components of the reverse cholesterol transport (RCT) pathway, in which cholesterol is removed from peripheral tissues and transferred to the liver for excretion. In the first step of the RCT pathway, lipid-poor apoA1 acts as an acceptor for cell cholesterol and phospholipids via the cell membrane protein ABCA1, generating nascent HDL. However, not all HDL is equivalent, and several studies have reported that individuals with coronary artery disease have HDL that is “dysfunctional” and no longer atheroprotective. The atherosclerotic lesion is a highly oxidative environment, and human lesions contain high levels of the antimicrobial enzyme myeloperoxidase (MPO), which we and others have shown can oxidize apoA1 and impair its function. We determined that the four tryptophan residues in human apoA1 (h-apoA1) are crucial in its MPO mediated loss of cholesterol acceptor function. We created a novel 4WF h-apoA1 variant, in which all four tryptophan residues are replaced by phenylalanine, which is resistant to MPO-mediated loss of function. We created and characterized transgenic mice that express high levels of the 4WF h-apoA1 isoform and found that these mice were resistant to inflammation. We also created human MPO transgenic mice, which over express MPO that can be further induced by zymosan treatment. Here we propose to characterize h-apoA1 modifications in mouse models and if these are modulated by inflammatory stimuli. We will also test whether the oxidant resistant 4WF apoA1 isoform can better protect from inflammation and sepsis, promote reverse cholesterol transport, delay atherosclerosis progression, and promote atherosclerosis regression in mice that over express MPO, creating an oxidative environment similar to that found in human lesions. Mechanistically, we will determine how the 4WF isoform protects mice from an acute phase response, and examine if the 4WF isoform better prevents myeloid cell proliferation and mobilization from the bone marrow leading to monocytosis and neutrophilia that are associated with atherosclerosis progression.

抽象的高密度脂蛋白 - 胆固醇（HDL-C）与心血管风险降低有关流行病学研究中的疾病（CVD）。尽管几种机制可能在HDL受保护中发挥作用效果，HDL及其主要蛋白质构建体载脂蛋白-AI（APOA1）是反向的关键组成部分胆固醇转运（RCT）途径，其中将胆固醇从外周组织去除并转移到排泄的肝脏。在RCT途径的第一步中，脂肪贫乏的apoA1充当细胞的受体通过细胞膜蛋白ABCA1胆固醇和磷脂，产生新生的HDL。但是，不是所有HDL都是等效的，几项研究报告说，冠状动脉疾病的个体患有 HDL是“功能失调”，不再是动脉保护性的。动脉粥样硬化病变是高度氧化的环境和人类病变含有高水平的抗菌酶髓过氧化物酶（MPO），我们和其他人已经显示的可能会氧化物ApoA1并损害其功能。我们确定这四个色氨酸保留在人apoA1（H-APOA1）中的MPO介导的胆固醇受体丧失至关重要功能。我们创建了一个新颖的4WF H-APOA1变体，其中所有四个色氨酸替代品均取代苯丙氨酸，对MPO介导的功能丧失具有抗性。我们创建并表征了转基因表达高水平的4WF H-APOA1同工型的小鼠，发现这些小鼠对炎。我们还创建了人类MPO转基因小鼠，这些小鼠超过了MPO，可以进一步由Zymosan治疗诱导。在这里，我们建议在鼠标模型和如果这些是由炎症刺激调节的。我们还将测试是否耐氧化剂4WF APOA1 同工型可以更好地保护感染和败血症，促进反向胆固醇运输，延迟动脉粥样硬化的进展，并促进过度表达MPO的小鼠动脉粥样硬化消退氧化物环境类似于人类病变中的氧化物环境。从机械上讲，我们将确定 4WF同工型保护小鼠免受急性相响应的侵害，并检查4WF同工型是否更好地预防髓样细胞的增殖和动员骨髓的动员，导致单核细胞增多和中性粒细胞增生与动脉粥样硬化进展有关。