Quantitative Assessment of HDL Function

本申请是为了响应PA-11-012-提高对HDL的认识- 该FOA的最终目标是开发可重复和可靠的测定HDL的方法， 功能和鉴定与HDL功能相关的新基因和途径。“我们相信我们有资格 为了满足这一需求，因为我们的研究计划侧重于了解导致 HDL的心脏保护作用。我们的方法的两个关键组成部分集中在（i）调查 控制胆固醇流出和HDL抑制巨噬细胞炎症的能力的因子，和（ii） 开发用于定量HDL中氧化产物和蛋白质的质谱方法。 重要的是，我们已经确定了可能损害HDL心脏保护作用的特定机制。 一个涉及髓过氧化物酶（MPO）对apoA-I（主要HDL蛋白）的氧化损伤。MPO损害 apoA-I通过ABCA 1途径从巨噬细胞中清除胆固醇的能力。另一个潜在 其机制涉及HDL携带的抗炎蛋白的改变。使用shotgun 蛋白质组学，我们已经证明，HDL携带一个独特的货物蛋白质在心血管疾病 (CVD)这些蛋白质可能对HDL的功能做出了以前未被怀疑的贡献。 此外，我们已经表明，患有CVD或急性炎症的受试者的HDL损害了他们的能力， 从巨噬细胞中清除胆固醇，这是胆固醇逆向转运的关键第一步。 我们的研究将利用三个独特的人群。第一组是控制组， 患有急性冠状动脉综合征的受试者和患有确诊CVD的受试者。第二个涉及控制 受试者和参加他汀类药物治疗前瞻性试验的CVD受试者。第三批人参加了 在一项对患有颈动脉疾病的受试者进行的前瞻性研究中， 通过MRI。这些宝贵资源的可用性将使我们能够调查具体的机制， 在患有CVD的受试者中产生功能失调的HDL，这是工业化社会中死亡的主要原因。

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.