Alterations in Unbound Free Fatty Acid Profiles in CVD

项目摘要-稳定性心血管疾病(CVD)患者的风险增加 第二个事件。对稳定的心血管疾病患者进行风险筛查最简单的方法是使用 生物标志物。然而，没有一个单一的标记物具有足够的预后能力。在这个项目中，我们将 确定游离脂肪酸(FFAu)的分布是否有助于预测糖尿病的风险 稳定性心血管疾病(CVD)患者的不良结局。血浆游离脂肪酸 长期以来，主要与白蛋白结合的游离脂肪酸(FFA)与心肌缺血有关。 虽然只存在NM浓度，但FFAU是生理上相关的部分 因为未绑定的和未绑定的被传输到细胞中。我们开发了荧光灯 用于测量FFAu的探针(ADIFAB)，发现FFAu水平与 TIMI II中PTCA和STEMI患者的心血管缺血。 由大约30个独特的FFA组成。因为它们有不同的生物效应，我们有 开发了一种确定不同FFAU(轮廓)的方法。NHLBI为我们提供了 来自和平试验的1200名患者的血浆(有临床数据)，该试验试图确定 血管紧张素转换酶抑制剂是否会改善心血管事件患者的预后。 我们使用我们的FFAU分析方法对200名PRESS患者进行了分析。对于大多数FFAU 病例(事件)与对照(无事件)之间没有显著差异。但是，对于未绑定的 α亚麻酸(αLNAu)病例平均摩尔分数是对照组的两倍多 (宝洁0.0002)。对200FFAU剖面中αLNAu浓度的ROC分析得出了C- 指数为0.702，优于目前最好的5个生物标志物的总和。这个结果 提示αLNAU可能是一个新的危险因素，反映了与当前事件垂直的事件 稳定性CVD的最佳生物标志物。出乎意料的是，两种结合能力最弱的FFAU，棕榈酸酯 (POAu)和亚麻酸(αLNAu)没有结合，尽管两者都存在结合 白蛋白。重要的是，我们发现POAu和α-LNAu在血液中发生了阻断解离 但在由白蛋白和FFA组成的水介质中不发生。在我们的FFAU配置文件中 健康血样和非心脏病患者未检测到POAu和α-LNAu。 这意味着在白蛋白中存在着POA和α-LNA解离的特异性抑制剂。 大多数血液样本，揭示了一种迄今未知的调节FFA代谢的机制 也是一个潜在的治疗靶点。因此，FFAU曲线揭示了看不见的生物效应 以总计FFA配置文件。在本项目中，我们将完成对数据的测量和分析 剩余的未分析的和平样品，并开始寻找释放的抑制物。

Project Summary- Patients with stable cardiovascular disease (CVD) have an elevated risk of a second event. The easiest method for screening risk in stable CVD patients would use biomarkers. However, no single marker has sufficient prognostic power. In this project we will determine whether profiles of unbound free fatty acids (FFAu) can help predict the risk of adverse outcomes in patients with stable cardiovascular disease (CVD). Plasma free fatty acids (FFA) which are mostly bound to albumin have long been associated with cardiac ischemia. Although present at only nM concentrations, the FFAu are the physiologically relevant fraction because the unbound and not the bound are transported into cells. We developed fluorescent probes (ADIFAB) to measure FFAu and found that FFAu levels were associated with cardiovascular ischemia during PTCA and in patients with STEMI in TIMI II. Human plasma is composed of about 30 unique FFA. Because they have distinct biological effects, we have developed a method to determine the different FFAu (profiles). The NHLBI provided us with plasma from 1200 patients (w/ clinical data) from the PEACE trial which sought to determine whether ACE inhibitors would improve outcomes in patients who had a cardiovascular event. We used our FFAu profiling method to profile 200 of the PEACE patients. For most of the FFAu there were no significant case (event) vs. control (no event) differences. However, for unbound alpha linolenate (αLNAu) the average case mole fractions were more than twice controls (p<0.0002). ROC analysis of the αLNAu concentrations in the 200 FFAu profiles yielded a C- index of 0.702, which is superior to the current 5 best biomarkers combined. This result suggests that αLNAu might be a novel risk factor that reflects events orthogonal to the current best biomarkers of stable CVD. Unexpectedly, two of the weakest binding FFAus, palmitoleate (POAu) and linolenate (αLNAu) had “zero” unbound, although both were present bound to albumin. Importantly, we found that blocking dissociation of POAu and α-LNAu occurs in blood but does not occur in aqueous media composed of albumin and FFA. In our FFAu profiles of healthy blood samples and non-cardiac diseases we found no measurable POAu and α-LNAu. This implies that there are specific inhibitors of POA and α-LNA dissociation from albumin in most blood samples, revealing a hitherto unknown mechanism for regulating FFA metabolism and a potential therapeutic target. Thus, FFAu profiles reveal biological effects that are invisible to total FFA profiles. In this project we will complete the measurement and analysis of the remaining unanalyzed PEACE samples and begin the search for the inhibitor of release.