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

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 在一般人群或选定的个人群体中诊断酒精滥用，例如 青少年和正在康复的酗酒者。因此，这项提议寻求开发诊断生物标记物 急性和慢性饮酒用于诊断高风险饮酒、检测复发的特征 饮酒，披露最近饮酒和怀孕等高风险情况。为此，研究正在进行 建议检查血清蛋白质和蛋白质模式，以寻找强大的非人类的潜在特征 灵长类动物模型不受共病药物使用、饮食不足和不可靠问题的困扰 对饮酒史的评估。在这些由NIAAA资助的、正在进行的、主题内的研究中，猴子 被诱使自愿大量饮酒。在研究过程中(包括 100只个体动物进行了多年的行为和观察)，血清样本通常被 收集并存档。建议进行实验，以筛选这些样本中潜在的生物标记物 然后被带到人类人口中。一种长期存在的非人灵长类动物的血清样本 自我给药研究将用作使用高通量进行生物标记物识别的训练集 蛋白质组学。样品将被处理以耗尽最丰富、最模糊的蛋白质，然后 TO-2-DGE(双向荧光差示凝胶电泳法)定量荧光鉴定 改变血清蛋白质表达，MALDI-ToF/ToF鉴定蛋白质种类。统计 验证将以盲目方式进行，使用来自独立群体的一组样本 自我管理的猴子，其中还将包含有关青少年脆弱性的数据。任何一种 假定的生物标志物将是敏感性(饮酒者中阳性分数的百分比)和特异性 (不饮酒人群中假阳性的百分比)。此外，这些研究将提供初步的 生物标志物签名的阳性预测值和阴性预测值的指数。 酒精滥用和酒精中毒的临床测试将有许多潜在的用途。发现蛋白质 酒精滥用和酒精中毒的生物标记物，来自受控的非人类灵长类种群自身的血清 服用乙醇将通过定量蛋白质组学方法进行检测。

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 processedto 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.