Novel MPO-derived plasma lipids as predictors of major adverse coronary events

描述（由申请人提供）：冠状动脉疾病是导致每年超过50万美国公民猝死的原因。在美国，开发预测主要不良冠状动脉事件风险增加的生物标志物是一项主要的公共卫生需求。血浆髓过氧化物酶（MPO）水平升高已被证明是急性冠状动脉综合征患者冠状动脉事件风险增加的预测因子。中性粒细胞释放MPO先于心肌损伤，并且已经表明MPO活性可能部分介导不良冠状动脉事件。因此，与血浆MPO水平相比，MPO产生的氧化产物的血浆水平可能是不良冠状动脉事件的更好预测因子。我们已经发现1-氯脂肪醛（1-ClOID）作为一种新的MPO产生的脂质氧化产物。在过去的一年中，我们已经发现1-氯脂肪酸（1-ClFA）和1-氯脂肪醇（1-ClFOH）是1-氯代维生素D的稳定代谢物，从中性粒细胞释放，也存在于人血浆中。因此，本提案的具体目的是检验血浆1-ClFA和1-ClFOH水平预测冠状动脉疾病患者主要不良冠状动脉事件风险增加的假设。克利夫兰临床基金会（Cleveland Clinic Foundation）一项名为Genebank的研究中的人血浆标本的广泛临床数据库和储存库将可用于这些研究。对于入组本研究的所有受试者，接受诊断性心脏导管插入术的受试者的大量临床表型数据均可用。完整的病史、实验室、血管造影和结局数据可用，受试者已同意使用这些标本和匿名临床数据，以支持心血管疾病相关的生化研究工作。病例队列设计将允许我们使用相同的对照人群来评估多种结局。病例定义将包括发生非致死性心肌梗死、卒中或死亡的受试者。对照组将从Genebank登记处的年龄和性别匹配的合格受试者的随机样本中抽取，这些受试者证明没有心血管疾病病史。在圣刘易斯大学，将使用电喷雾电离在三重四极杆仪器上通过LC-MS定量血浆1-ClFA。血浆1-ClFOH将在衍生为其五氟苯甲酰基酯后，使用GC-MS和负离子化学电离进行定量。通过与合成制备的氘代内标物进行比较，对每种分析物进行定量。将这些新生物标志物的水平与先前在这些标本中定量的其他生物标志物（包括C反应蛋白、MPO、氯酪氨酸、硝基酪氨酸和肌钙蛋白T）的水平进行比较。Genebank中这些收集的人类样本的可用性提供了一个非凡的机会，以确定这些新的MPO催化的氧化产物在血浆中的一个重点和明确的心脏病患者组作为新的生物标志物。此外，这项研究将首次直接测量全身MPO催化的脂质氧化，并评估其与心血管疾病的潜在临床相关性。公共卫生相关性：大约有1300万美国公民患有冠状动脉疾病，这是男性和女性死亡的主要原因。在本研究中，我们将检验新的MPO产生的脂质氧化产物的血浆水平预测主要不良冠状动脉事件的假设。识别这些新的脂质氧化产物作为未来主要不良冠状动脉事件的预测因子有可能显著影响冠状动脉疾病患者的护理和结局。

DESCRIPTION (provided by applicant): Coronary artery disease is responsible for the sudden death of over 500,000 U.S. citizens per year. The development of biomarkers that predict increased risk of major adverse coronary events is a major public health need in the United States. Elevated plasma myeloperoxidase (MPO) levels have been shown to be a predictor of increased risk of coronary events in patients with acute coronary syndromes. MPO release from neutrophils precedes myocardial injury, and it has been suggested that MPO activity may mediate, in part, adverse coronary events. Accordingly, in comparison to plasma MPO levels, plasma levels of MPO-generated oxidation products may be better predictors of adverse coronary events. We have discovered 1-chloro-fatty aldehyde (1- ClFALD) as a novel MPO-generated lipid oxidation product. In the past year, we have revealed that 1-chloro- fatty acid (1-ClFA) and 1-chloro-fatty alcohol (1-ClFOH) are stable metabolites of 1-ClFALD that are released from neutrophils and are also present in human plasma. Accordingly, the specific aim of this proposal is to test the hypothesis that plasma levels of 1-ClFA and 1-ClFOH predict increased risk of major adverse coronary events in patients with coronary artery disease. An extensive clinical database and repository of human plasma specimens from a study called Genebank at the Cleveland Clinic Foundation will be available for these studies. Extensive clinical phenotypic data from subjects undergoing diagnostic cardiac catheterization are available for all subjects enrolled in this study. Complete medical history, laboratory, angiographic and outcomes data are available, and subjects have provided consent for use of these specimens and anonymonized clinical data to support biochemical research efforts related to cardiovascular disease. A case-cohort design will allow us to evaluate multiple outcomes using the same control population. The case definition will include subjects that suffered non-fatal myocardial infarction, stroke or death. Controls will be drawn from a random sample of age and gender-matched eligible subjects from the Genebank registry demonstrating no inter- vening history of cardiovascular disease. At Saint Louis University, plasma 1-ClFA will be quantified by LC-MS on a triple quadrupole instrument using electrospray ionization. Plasma 1-ClFOH will be quantified using GC- MS with negative ion-chemical ionization following derivatization to its pentafluorobenzoyl ester. Each analyte will be quantified by comparisons to synthetically-prepared deuterated internal standards. Comparisons will be made between levels of these new biomarkers to that of other biomarkers including C-reactive protein, MPO, chlorotyrosine, nitrotyrosine and troponin T that have previously been quantified in these specimens. The availability of these collected human samples in Genebank provides an extraordinary opportunity to identify these novel MPO-catalyzed oxidation products in the plasma of a focused and well-defined group of heart patients as novel biomarkers. Furthermore, this study will allow a direct measure of systemic MPO-catalyzed lipid oxidation for the first time and evaluation of its potential clinical relevance to cardiovascular disease. PUBLIC HEALTH RELEVANCE: About 13 million U.S. citizens have coronary artery disease, and it is the leading cause of death among both men and women. In the proposed studies, we will test the hypothesis that plasma levels of novel MPO-generated lipid oxidation products predict major adverse coronary events. Identifying these novel lipid oxidation products as predictors of future major adverse coronary events has the potential to significantly impact care and outcomes of patients with coronary artery disease.