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

描述(申请人提供)：酒精滥用和酗酒仍然是非常严重的社会问题。一个重大问题是，无论是在一般人群中，还是在特定的个人群体中，如青少年和正在康复的酗酒者，都无法诊断酒精滥用。因此，该提案寻求开发急性和慢性饮酒的诊断生物标记物标记，用于诊断高风险饮酒、检测再次饮酒、披露最近饮酒和怀孕等高风险情况下的饮酒情况。为此，建议进行研究，在一个强大的非人类灵长类动物模型中检查血清蛋白质和蛋白质模式，以寻找潜在的特征，该模型不会受到并存药物使用、饮食不足和不可靠的饮酒史评估等问题的阻碍。在这些由NIAAA资助的、正在进行的受试者内部研究中，猴子被诱导自愿饮用大量酒精。在研究过程中(包括100多只动物的多年行为和观察)，血清样本被常规收集和存档。有人提议进行实验，以筛选这些样本中的潜在生物标记物，然后将其带入人类群体。来自长期的非人类灵长类自我给药研究的血清样本将被用作使用高通量蛋白质组学识别生物标记物的训练集。样品将被处理以去除最丰富的、模糊的蛋白质，然后进行2-DGE(2-D荧光凝胶内差异电泳法)以定量荧光鉴定改变的血清蛋白质表达，然后进行MALDI-ToF/ToF蛋白质种类鉴定。统计验证将以盲目的方式进行，使用来自独立的自我管理猴子群体的一组样本进行统计验证，其中也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都将是敏感性(饮酒者中阳性分数的百分比)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记物签名提供积极和消极预测值的初始指数。酒精滥用和酒精中毒的临床测试将有许多潜在的用途。为了发现酒精滥用和酒精中毒的蛋白质生物标记物，将用定量蛋白质组学方法检测自服酒精的受控非人类灵长类种群的血清。

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.