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Biophysics of HDL Dysfunction

HDL 功能障碍的生物物理学

基本信息

批准号：
8511809
负责人：
Kirkwood Arthur Pritchard
金额：
$ 62.81万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-16 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8511809
关键词：
ATP binding cassette transporter 1 ATP-Binding Cassette Transporters Affect Affinity Anti-Inflammatory Agents Anti-inflammatory Antioxidants Apolipoprotein A-I Atherosclerosis Automation Binding Biological Assay Biophysics Biosensor Blinded Cardiovascular Diseases Carrier Proteins Cells Cholesterol Cholesterol Esters Cholesterol Homeostasis Clinical Clinical Research Complex Cultured Cells Development Diagnosis Diagnostic Diagnostic tests Effectiveness Enzymes Esterification Event Family Foundations Functional disorder Genetic Engineering Goals Healthcare Heart Diseases High Density Lipoprotein Cholesterol High Density Lipoproteins Hospitals Human In Vitro Individual Inflammation Inflammatory Interferometry Knowledge Label Laboratories Lecithin Lipids Lipoprotein Binding Lipoproteins Liver Low Density Lipoprotein oxidation Manuals Measurement Measures Mediating Metabolism Modeling Modification Molecular Mus Myocardial Infarction Oxidative Stress Pathway interactions Patients Peroxidases Pharmaceutical Preparations Phospholipids Physiological Platelet Activating Factor Play Property Protective Agents Proteins Publishing Reactive Oxygen Species Reporting Research Risk Role SR-B proteins Sampling Series Source Stroke Techniques Testing Time Transferase Validation Vascular Diseases Xanthine Oxidase aryldialkylphosphatase base case control cost design experience glutathione peroxidase heart disease risk high density lipoprotein-1 high density lipoprotein-2 high density lipoprotein-3 improved in vitro Assay in vitro Bioassay indexing insight novel oxidation oxidized lipid prevent public health relevance receptor reconstitution research study reverse cholesterol transport scavenger receptor uptake

项目摘要

DESCRIPTION (provided by applicant): The oxidation status of HDL plays an important role in determining how this lipoprotein prevents or promotes atherosclerosis. Since patients with normal levels of HDL experience atherogenic events (ie., stroke, myocardial infarction), HDL function itself may be a stronger indicator of cardiovascular disease than the actual levels of HDL cholesterol. Bioassays of HDL function are complex, time-consuming, technician- and technique-dependent and difficult to reproduce between labs. Thus, highly efficient assays of HDL function are desperately needed. In this application, we hypothesize that the biophysics of HDL interactions with biomolecules the mediate HDL-dependent cholesterol metabolism represent a novel source of physiological information that can be used to assess the functional state of HDL. Using biolayer interferometry (BLI), a new label-free technique for measuring biomolecular interactions, we will develop a series of novel assays of HDL "function" to determine if and the extent to which oxidation impacts on the ability of HDL to interact with 1) anti- and pro-inflammatory enzymes (paraoxonase [PON1], platelet activating factor acetylhydrolase [PAF-AH], myeloperoxidase [MPO] and xanthine oxidase [XO]) to prevent LDL oxidation; 2) or ATP binding cassette (ABC) transporter 1 (ABCA1)/ABCG1, lecithin cholesteryl acyl transferase (LCAT), cholesteryl ester transfer protein (CETP) and scavenger receptor class B1 (SR-B1) to mediate HDL-cholesterol release, esterification, transfer and uptake, respectively. The mechanisms by which HDL is anti-inflammatory or participates in reverse cholesterol transport is directly dependent on the ability of HDL to bind these biomolecules. As such, any oxidation-induced changes in HDL binding affinity for these biomolecules should provide a sensitive index of HDL functionality. In Aim 1, we propose to measure the extent to which oxidized lipids and proteins in reconstituted HDL (r-HDL) impair HDL function using established in vitro bioassays of cholesterol release, esterification, transfer and uptake. In Aim 2, we will ue BLI assays to determine how oxidized lipids and proteins in r-HDL alter binding rates and affinity for the proteins and enzymes that mediate HDL's ability to inhibit LDL oxidation or promote specific steps in the reverse cholesterol transport pathway. In Aim 3, we propose to verify and validate that BLI assays by 1) correlating BLI assays with bioassays of HDL function in established murine models of vascular disease and 2) by using BLI assays to predict which patients have clinically-diagnosed atherosclerosis in case-controlled, blinded human studies. Successful completion of these studies will lay the foundation for the development of a new clinical assay for determining HDL functionality. Validation of these new assays will allow us to identify which patients have dysfunctional HDL in a time frame and for a cost that is compatible with clinical reference laboratories. Development of these novel assays will make it possible, for the first time, to perform large clinical studies to fully test the idea that dysfunctional HDL is better indicator of atherosclerotic risk.