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， 诊断一般人群或选定群体中的酒精滥用，例如 青少年和正在戒酒的人因此，该提议寻求开发诊断生物标志物， 急性和慢性酒精消费的特征，用于诊断高危饮酒，检测复发， 饮酒，披露最近饮酒和怀孕等高风险情况。为此，研究 建议检查血清蛋白和蛋白质模式的潜在签名，在一个强大的非人类 灵长类动物模型，不受共病药物使用，饮食不足和不可靠的问题的阻碍 饮酒史的评估。在这些NIAAA资助的、正在进行的受试者内研究中，猴子 被诱导自愿喝大量的酒。在研究过程中（包括 100只个体动物，涵盖多年的行为和观察），血清样本已常规 收集并存档。提出了实验来筛选这些样品的潜在生物标志物， 然后被带到人类中。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志物鉴定的训练集 蛋白质组学样本将被处理以去除最丰富的模糊蛋白质， 2-DIGE（2-D荧光差异凝胶电泳）用于定量荧光鉴定 改变的血清蛋白质表达，随后进行蛋白质种类的MALDI-ToF/ToF鉴定。统计 将使用来自独立菌落的样品测试集以盲法进行验证， 自我管理的猴子，其中也将载有关于青少年脆弱性的数据。任何一个关键标准 推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 （非饮酒人群中假阳性的百分比）。此外，这些研究将提供初步的 生物标志物特征的阳性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 乙醇滥用和酒精中毒的生物标志物，来自对照非人灵长类群体的血清， 将通过定量蛋白质组学方法来检查给予乙醇。

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.