Structure-Function Analysis of Triglyceride Regulators ApoC-III and ApoA-V Using Natural Variants

使用天然变体对甘油三酯调节剂 ApoC-III 和 ApoA-V 进行结构-功能分析

基本信息

批准号：
9306180
负责人：
Daniel James Rader
金额：
$ 39.6万
依托单位：
UNIVERSITY OF PENNSYLVANIA
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：
2016-07-01 至 2020-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9306180
关键词：
APOA5 gene Adopted Affect Amino Acids Apolipoprotein A-I Apolipoprotein E Apolipoproteins Apolipoproteins A Apolipoproteins B Apolipoproteins C Area Atherosclerosis Biochemical Blood CETP gene Cardiovascular Diseases Cause of Death Code Coronary Arteriosclerosis Dependovirus Deuterium Development Disease Effectiveness Enzymes Etiology Fatty Acids Genes Genetic study Hepatic Human Human Genetics Hydrogen Hypertriglyceridemia In Vitro Industrialization Investigation Knockout Mice LDL Cholesterol Lipoproteins Lead Lipid Binding Lipids Lipoprotein Binding Lipoproteins Maps Mass Spectrum Analysis Mediating Metabolism Modeling Modernization Molecular Mus Mutation Myocardial Infarction Patients Physiological Plasma Play Prevalence Production Property Proteins Recombinants Resolution Risk Risk Factors Role Structure Structure-Activity Relationship Testing Tissues Transgenic Mice Transgenic Organisms Triglyceride Metabolism Triglycerides Variant apoB mRNA editing catalytic subunit apolipoprotein C-III apolipoprotein E-3 apolipoprotein E-4 atherogenesis base burden of illness cardiovascular disorder risk cardiovascular risk factor disorder risk effective therapy experimental study humanized mouse in vivo insight lipoprotein lipase lipoprotein lipase activators lipoprotein lipase inhibitor loss of function mortality mouse model mutant novel novel strategies protein folding protein function targeted treatment uptake

项目摘要

PROJECT SUMMARY Coronary artery disease (CAD) remains among the highest disease burdens in the modern world. Despite effective treatments focused on specific risk factors such as LDL cholesterol (LDL-C), prevalence and mortality from this disease remains very high. This necessitates investigating additional risk factors and identifying novel targets for treatment. Apolipoproteins C-III and A-V are reciprocal regulators of TG in plasma and were recently implicated through human genetics as being significantly related to risk of developing CAD. Genetic studies suggested that inactivating ApoC-III or enhancing ApoA-V, may protect against CAD. However, we currently have very little understanding of the structure and mechanisms of these two proteins and how the disease-associated mutations exert their effects to regulate TG levels and CAD risk. In this proposal, we seek to elucidate the helical and structural dynamics of the wild-type (WT) forms of these two apolipoproteins in the lipid-free and lipid-bound states through hydrogen-deuterium exchange and mass spectrometry to amino acid- level resolution. We will also study the effects the identified coding mutations on protein folding, dynamics, and lipid binding. We posit that the identified mutations influence lipid binding and cause altered TG metabolism through impacting lipoprotein association, LPL activity, TG-rich lipoprotein clearance and hepatic TG metabolism. We will test hypotheses for how each variant is acting through biochemical approaches and complementary in vivo studies comparing expression of WT and mutant ApoC-III/A-V in humanized mouse models. Finally, we will assess the impact of the CAD-associated ApoC-III/A-V mutations on atherogenesis in vivo. The proposed experiments will decipher the specific mechanisms of action of ApoC-III and ApoA-V in regulating TG metabolism and risk of CAD, and determine the structural features most crucial to these functions.

项目概要冠状动脉疾病（CAD）仍然是现代世界负担最重的疾病之一。尽管针对特定危险因素的有效治疗，例如低密度脂蛋白胆固醇 (LDL-C)、患病率和死亡率这种疾病的发病率仍然很高。这需要调查额外的风险因素并确定新的治疗目标。载脂蛋白 C-III 和 A-V 是血浆中 TG 的相互调节因子，最近通过人类遗传学发现与患 CAD 的风险显着相关。遗传研究表明，灭活 ApoC-III 或增强 ApoA-V 可以预防 CAD。然而，我们目前对这两种蛋白质的结构和机制以及如何发挥作用还知之甚少。疾病相关突变发挥其调节 TG 水平和 CAD 风险的作用。在本提案中，我们寻求阐明这两种载脂蛋白野生型 (WT) 形式的螺旋和结构动力学通过氢-氘交换和质谱分析氨基酸-无脂质和脂质结合状态级别分辨率。我们还将研究已识别的编码突变对蛋白质折叠、动力学和脂质结合。我们假设已确定的突变影响脂质结合并导致 TG 代谢改变通过影响脂蛋白结合、LPL 活性、富含 TG 的脂蛋白清除率和肝脏 TG 代谢。我们将通过生化方法测试每种变体如何发挥作用的假设比较 WT 和突变型 ApoC-III/A-V 在人源化小鼠中表达的互补体内研究模型。最后，我们将评估 CAD 相关的 ApoC-III/A-V 突变对动脉粥样硬化形成的影响体内。拟议的实验将破译 ApoC-III 和 ApoA-V 的具体作用机制调节 TG 代谢和 CAD 风险，并确定对这些最重要的结构特征功能。