Serum Biomarkers of Alcohol Self-Administration in Non-Human Primates

描述（由申请人提供）：酒精滥用和酗酒仍然是非常严重的社会问题。一个重要的问题是无法诊断一般人群或特定个人群体（如青少年和戒酒者）的酒精滥用情况。因此，本提案旨在开发急性和慢性饮酒的诊断生物标志物特征，以诊断高风险饮酒，检测饮酒复发，披露近期饮酒以及怀孕等高风险情况。为此，研究人员建议在一个强大的非人类灵长类动物模型中检查血清蛋白和蛋白质模式，以寻找潜在的特征，该模型不受共病药物使用、饮食不足和不可靠的饮酒史评估等问题的阻碍。在这些niaaa资助的、正在进行的研究中，猴子被诱导自愿喝大量的酒。在研究过程中（涉及100多只个体动物，涵盖多年的行为和观察），血清样本已被常规收集并存档。实验建议筛选这些样本，寻找潜在的生物标志物，然后将其应用于人类群体。一项长期进行的非人类灵长类动物自我给药研究的血清样本将被用作高通量蛋白质组学生物标志物鉴定的训练集。对样品进行处理，去除最丰富的模糊蛋白，然后进行2-DIGE （2-D荧光差异凝胶电泳）定量荧光鉴定改变的血清蛋白表达，然后进行MALDI-ToF/ToF鉴定蛋白质种类。统计验证将以盲法进行，使用一组来自一群独立的自我管理猴子的样本进行测试，其中还将包含有关青少年脆弱性的数据。任何假定的生物标志物的关键标准将是敏感性（饮酒者中阳性得分的百分比）和特异性（非饮酒者中假阳性的百分比）。此外，这些研究将为生物标志物特征提供阳性和阴性预测值的初始指标。对乙醇滥用和酒精中毒的临床试验将有许多潜在的用途。为了发现乙醇滥用和酒精中毒的蛋白质生物标志物，将用定量蛋白质组学方法检测受控的非人类灵长类动物群体自我施用乙醇的血清。

DESCRIPTION (provided by applicant): Ethanol abuse and alcoholism remain very serious societal problems. A significant problem is the inability to diagnose alcohol abuse either in the general population or within selected groups of individuals such as adolescents and the recovering alcoholic. Accordingly, this proposal seeks to develop diagnostic biomarker signatures of acute and chronic alcohol consumption for diagnosing high-risk drinking, detecting relapse to drinking, disclosing recent drinking and in high risk situations such as pregnancy. To this end, studies are proposed to examine serum proteins and protein patterns for potential signatures in a powerful non-human primate model that is not encumbered by problems of comorbid drug use, inadequate diet and unreliable assessments of drinking history. In these NIAAA-funded, ongoing, within-subject studies, monkeys have been induced to voluntarily drink large amounts of alcohol. In the course of the studies (encompassing over 100 individual animals covering years of behavior and observation), serum samples have routinely been collected and archived. Experiments are proposed to screen these samples for potential biomarkers that can then be taken forward into the human population. Serum samples from a long-standing nonhuman primate self-administration study will be used as a training set for biomarker identification using high throughput proteomics. Samples will be processed to deplete the most abundant, obscuring proteins and then subjected to 2-DIGE (2-D Fluorescence Difference In-Gel Electrophoresis) for quantitative fluorescence identification of altered serum protein expression followed by MALDI-ToF/ToF identification of protein species. Statistical validation will be conducted, in a blinded fashion, using a test set of samples from an independent colony of self-administering monkeys, which will also contain data on adolescent vulnerability. The key criteria of any putative biomarkers will be sensitivity (percentage of positive scores among drinkers) and specificity (percentage of false positives in a non-drinking population). In addition, these studies will provide initial indices of positive and negative predictive values for biomarker signatures. A clinical test for ethanol abuse and alcoholism would have many potential uses. To discover protein biomarkers of ethanol abuse and alcoholism, serum from a controlled non-human primate population self- administering ethanol will be examined by quantitative proteomic methods.