Lipoprotein Density Profiling for Clinical Studies

用于临床研究的脂蛋白密度分析

• 批准号: 6610161
• 负责人: RONALD D MACFARLANE
• 金额: $ 21.83万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-15 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6610161
• 关键词: analytical method; atherosclerosis; blood lipid; blood lipoprotein metabolism; cardiovascular disorder diagnosis; cardiovascular disorder risk; cesium; cholesterol; clinical research; density gradient ultracentrifugation; high density lipoproteins; human tissue; low density lipoprotein; metal complex; method development; very low density lipoprotein

DESCRIPTION (provided by applicant): The broad, and long-term objective is to have the lipoprotein particle density profile be part of an individual's risk assessment for cardiovascular disease (CVD). Lipoproteins are nanoparticles circulating in the blood that mediate lipid metabolism. They are divided into 3 major classes based on their densities:very-low density lipoprotein(VLDL), low density lipoprotein (LDL), and high density lipoprotein (HDL). The cholesterol content of LDL, (LDL-C) correlates positively with the risk of cardiovascular disease (CVD) while the cholesterol content of HDL, (HDL-C) is inversely related. Subclasses based on density are associated with increased risk of CVD. These include remnant lipoprotein, lipoprotein(a), dense LDL and dense HDL. An individual's lipoprotein particle density profile is a composite of relative risk factors linked to the development of arterial atherosclerosis. Density gradient ultracentrifugation (DGU) has long been considered to be the fundamental method for the characterization, isolation and quantitation of lipoproteins. It would also be the ideal method for measuring an individual's lipoprotein particle density profile in the assessment of relative risk for CVD. But, experimental problems associated with this method, since it was first used in this application a half-century ago, diminished its general utility in clinical chemical analysis. The tabletop very fast ultracentrifuge, introduced in the past decade, has transformed the analysis to microscale operation and analysis time from days to hours. However, difficulties in achieving high precision in sample preparation and the limited choices of density-gradient forming solutes (alkali halides, sugars) remain as problematic for the adoption of DGU for large-scale lipoprotein analysis. The "perfect solute" would be one that can form a reproducible density gradient from a homogeneous solution in a high g-force environment and allow lipoprotein particles to focus at their densities in a short time frame. The novel feature of this proposal is the introduction of a new class of density-gradient forming solutes (metal ion complexes) that gives versatility at the molelcular structure level in influencing sedimentation and diffusion rates in the ultracentrifuge. The Cs+ salt of Bi(EDTA) has been chosen. Preliminary studies suggest that it may be the "perfect solute". The specific aims focus on fundamental studies using Cs(BiEDTA) to achieve the high accuracy and precision required for clinical applications in measuring the lipoprotein particle density profile. A second specific aim is to use this same profile to isolate nd quantitate cholesterol levels of LDL-C, and HDL-C and evaluate accuracy by comparision with CDC-methods. A third specific aim is to link DGU with immunoreactivity to identify apolipoprotein-specific components of the liprotein particle density profile.

描述（由申请人提供）：广泛的和长期的目标是使脂蛋白颗粒密度概况成为个人心血管疾病风险评估（CVD）的一部分。脂蛋白是纳米颗粒，在介导脂质代谢的血液中循环。它们根据其密度分为3个主要类别：非常低的密度脂蛋白（VLDL），低密度脂蛋白（LDL）和高密度脂蛋白（HDL）。 LDL（LDL-C）的胆固醇含量与心血管疾病（CVD）的风险正相关，而HDL（HDL-C）的胆固醇含量成反比。 基于密度的子类与CVD风险增加有关。这些包括残留的脂蛋白，脂蛋白（A），致密的LDL和密集的HDL。个体的脂蛋白颗粒密度谱是与动脉动脉粥样硬化发展有关的相对风险因素的综合。 长期以来，密度梯度超速离心（DGU）长期以来被认为是脂蛋白表征，分离和定量的基本方法。在评估CVD相对风险中，这也是测量个体脂蛋白颗粒密度谱的理想方法。但是，与此方法相关的实验问题，因为它是在半个世纪前首次在该应用中使用的，因此在临床化学分析方面的一般效用降低了。在过去的十年中引入的桌面非常快速的超级离心设备已将分析转化为微观的操作和分析时间从几天到小时。然而，在样品制备中获得高精度和密度梯度形成溶质的有限选择（碱卤化物，糖）的困难仍然是用于大规模脂蛋白分析的DGU的问题。 “完美的溶质”将是可以在高g强环境中从均匀溶液中形成可再现的密度梯度的一种，并使脂蛋白颗粒在短时间内聚焦在其密度上。 该提案的新特征是引入新的密度梯度形成溶质（金属离子络合物），该溶质（金属离子络合物）在影响超级离心机的沉积和扩散速率方面具有多功能性。已经选择了BI（EDTA）的CS+盐。初步研究表明，这可能是“完美的溶质”。该特定目的专注于使用CS（BIEDTA）的基本研究，以实现测量脂蛋白颗粒密度谱的临床应用所需的高精度和精度。第二个具体目的是使用相同的配置文件来分离ND定量LDL-C和HDL-C的胆固醇水平，并通过与CDC方法进行比较来评估准确性。第三个具体目的是将DGU与免疫反应性联系起来，以识别脂蛋白颗粒密度谱的载脂蛋白特异性成分。