Proteomic Biosignatures of Chronic Drug Exposure

描述(由申请人提供)：基于长期血浆蛋白变化的生物签名可用于识别易重复使用药物的患者，从而促进早期干预。我们将利用大鼠长期接触成瘾药物和差异蛋白质组表达来测试寻找这样一个特征的可行性。根据14天的递增剂量计划(慢性暴露)，大鼠将接受可卡因或非成瘾性多巴胺转运体抑制剂或吗啡或生理盐水的治疗，并在最后一次药物治疗后收集最多75天的血浆。蛋白质组学分析将包括逐步耗尽丰富的血浆蛋白，然后在线液相色谱-质谱仪分析由非常低丰度的蛋白质组成的耗尽的蛋白质组和中等丰度的结合蛋白。其改变的表达可保持数周，例如30天，并最终恢复到基线的蛋白质将是候选生物标记物，可用于在成瘾药物不再容易测量的时候检测最近的暴露。最佳候选者将通过不需要蛋白质抗体的定量多重分析中的多反应监测来验证。蛋白质组的变化将被检查与慢性药物治疗的行为和神经化学后遗症的时间相关性，这将被平行测量。将使用统计分析和数据挖掘来识别具有所需特性的生物签名。一项成功的可行性研究将激发和启发对人类受试者样本的类似研究。 与公共卫生相关：对分析生物标记物的需求尚未得到满足，这是慢性药物暴露的生物特征，将有助于识别易重复用药的患者。一项有效的测试可能会成为常规医疗保健的一部分，以确定需要治疗药物使用障碍的患者，并减轻吸毒成瘾的个人和社会负担。

DESCRIPTION (provided by applicant): A biosignature based on long-term plasma protein changes specifically associated with chronic drug exposure can be used to identify patients predisposed to repeated drug use and thereby facilitate early intervention. We will test the feasibility of finding such a signature using chronic exposure of rats to addictive drugs and differential proteomic expression. Rats will receive treatment with cocaine or a non-addictive dopamine transporter inhibitor, or morphine, or saline according to a 14 day escalating dose schedule (chronic exposure) and plasma collected up to 75 days following last drug treatment. Proteomic analysis will involve stepwise depletion of abundant plasma proteins followed by liquid chromatography coupled online with mass spectrometry to analyze both the depleted proteome consisting of very low abundance proteins and the bound proteins of intermediate abundance. Proteins whose altered expression is maintained for several weeks, e.g. 30 days, and eventually return to baseline would be candidate biomarkers that could be used to detect recent exposure at a time when the addictive drug can no longer be readily measured. Top candidate will be validated by multiple reaction monitoring in a quantitative multiplexed assay that does not require antibodies to the proteins. Proteomic changes will be examined for temporal correlation with behavioral and neurochemical sequelae of chronic drug treatment that will be measured in parallel. Statistical analysis and data mining will be used to identify a biosignature with the desired properties. A successful feasibility study would stimulate and inform similar studies on human subject samples. PUBLIC HEALTH RELEVANCE: There is an unmet need for analytical biomarkers, a biosignature, of chronic drug exposure that would help to identify patients predisposed to repeated drug. An effective test could become part of routine medical care that would identify patients in need of treatment for substance use disorders and mitigate the personal and societal burdens of drug addiction.