Quantitative Assessment of HDL Function

本申请是对PA-11-012的响应--旨在提高对HDL语言的理解- 其中写道：“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。 为了满足这一需求，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。我们方法的两个关键部分集中在(I)调查 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损害 载脂蛋白A-I通过ABCA1途径从巨噬细胞中清除胆固醇的能力。另一种潜力 其机制涉及高密度脂蛋白携带的抗炎蛋白的改变。使用猎枪 蛋白质组学，我们已经证明了高密度脂蛋白在心血管疾病中携带着独特的蛋白质货物 (CVD)受试者，这些蛋白质可能对高密度脂蛋白的功能做出了以前未被怀疑的贡献。 此外，我们还表明，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损害的能力。 从巨噬细胞中清除胆固醇，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。第一个涉及对照对象， 有急性冠脉综合征的受试者和既有心血管疾病的受试者。第二个问题涉及控制 受试者和患有心血管疾病的受试者参加了他汀类药物治疗的前瞻性试验。第三批人参加了 在一项对颈动脉疾病受试者进行动脉粥样硬化进展评估的前瞻性研究中 通过核磁共振检查。这些宝贵资源的可获得性将使我们能够调查 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，心血管疾病是工业化社会的主要死亡原因。

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 exhibts 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.