Quantitative Assessment of HDL Function

描述(由申请人提供)：本申请是响应PA-11-012--旨在更好地理解高密度脂蛋白--的要求而提交的，该申请声明：“本FOA的最终目标是开发可重复性和健壮的分析方法来测量高密度脂蛋白功能，并确定与高密度脂蛋白功能相关的新基因和途径。”我们相信我们完全有资格满足这一需求，因为我们的研究计划侧重于了解影响高密度脂蛋白的心脏保护作用的因素。我们方法的两个关键部分集中在(I)研究控制胆固醇外流的因素和高密度脂蛋白抑制巨噬细胞炎症的能力，以及(Ii)开发用于定量高密度脂蛋白中氧化产物和蛋白质的质谱学方法。重要的是，我们已经确定了可能损害高密度脂蛋白心脏保护作用的具体机制。一种是髓过氧化物酶(MPO)对载脂蛋白A-I(主要的高密度脂蛋白)的氧化损伤。MPO通过ABCA1途径损害apoA-I从巨噬细胞中清除胆固醇的能力。另一种可能的机制涉及高密度脂蛋白携带的抗炎蛋白的变化。利用鸟枪式蛋白质组学，我们已经证明了在心血管疾病(CVD)受试者中，高密度脂蛋白携带着一种独特的蛋白质，这些蛋白质可能对高密度脂蛋白的功能做出了以前意想不到的贡献。此外，我们已经证明，患有心血管疾病或急性炎症的受试者的高密度脂蛋白表现出从巨噬细胞中清除胆固醇的能力受损，这是反向胆固醇运输的关键第一步。我们的研究将利用三个独特的人类群体。第一组涉及对照受试者、急性冠脉综合征受试者和既往心血管疾病受试者。第二项研究涉及对照受试者和接受他汀类药物治疗的CVD受试者。第三组人群参与了一项针对颈动脉疾病受试者的前瞻性研究，该研究通过MRI对动脉粥样硬化进展进行了评估。这些宝贵资源的可获得性将使我们能够研究在患有心血管疾病的受试者中产生功能失调的高密度脂蛋白的具体机制，心血管疾病是工业化社会的主要死亡原因。 与公共卫生相关：识别改变高密度脂蛋白功能的蛋白质，并在有心血管疾病风险的受试者中选择性地富含或耗尽，将支持炎症将人类高密度脂蛋白转化为功能失调的假说。我们的长期目标是了解影响高密度脂蛋白去除胆固醇和抑制巨噬细胞炎症能力的因素，这可能对高密度脂蛋白的治疗具有重要意义。

DESCRIPTION (provided by applicant): This application is submitted in response to PA-11-012-Toward an improved understanding of HDL- which states "The ultimate goal of this FOA is to develop reproducible and robust assays to measure HDL function and to identify novel genes and pathways related to HDL function." We believe we are well-qualified to address this need because our research program has focused on understanding the factors that contribute to the cardioprotective effects of HDL. Two key components of our approach have centered on (i) investigating the factors that control cholesterol efflux and HDL's ability to inhibit macrophage inflammation, and (ii) developing mass spectrometric approaches for quantifying oxidation products and proteins in HDL. Importantly, we have identified specific mechanism that may impair the cardioprotective effects of HDL. One involves oxidative damage of apoA-I (the major HDL protein) by myeloperoxidase (MPO). MPO impairs apoA-I's ability to remove cholesterol from macrophages by the ABCA1 pathway. Another potential mechanism involves alterations in the anti-inflammatory proteins that are carried by HDL. Using shotgun proteomics, we have demonstrated that HDL carries a unique cargo of proteins in cardiovascular disease (CVD) subjects and that those proteins might make previously unsuspected contributions to HDL's function. Moreover, we have shown that HDL of subjects with CVD or acute inflammation exhibits impaired ability to remove cholesterol from macrophages, the key first step in reverse cholesterol transport. Our studies will take advantage of three unique human populations. The first involves control subjects, subjects with acute coronary syndrome, and subjects with established CVD. The second involves control subjects and subjects with CVD enrolled in a prospective trial of statin therapy. The third population took part in a prospective study of subjects with carotid artery disease that was evaluated for atherosclerotic progression by MRI. The availability of these valuable resources will enable us to investigate specific mechanisms for generating dysfunctional HDL in subjects with CVD, the leading cause of death in industrialized societies. PUBLIC HEALTH RELEVANCE: Identifying proteins that alter HDL's function and are selectively enriched or depleted in subjects at risk for CVD would support the hypothesis that inflammation converts human HDL to a dysfunctional form. Our long-term goal is to understand the factors that impair HDL's ability to remove cholesterol and inhibit macrophage inflammation, which may have important implications for HDL therapeutics.