Using Phage Display to Identify Novel Peptide Modulators of Ethanol Targets

描述(申请人提供)：乙醇是一种被广泛使用的滥用药物，其作用机制复杂且鲜为人知。理解酒精作用机制的一个主要障碍是大量假定的乙醇靶标。虽然已知酒精对许多细胞成分有影响，但每个靶点在酒精对整体生理和行为影响中的贡献尚不清楚。我们提出了一种在不改变野生型系统的情况下分离乙醇对单个目标的影响的方法。我们计划利用噬菌体展示技术来识别与单个假定的乙醇靶标-甘氨酸受体(GlyR)具有高特异性结合的多肽。然后将对这些多肽进行电生理学测试，以确定其对GlyR的调节作用。这项拟议工作的目标是在不影响其他乙醇靶点的情况下，识别模拟或拮抗乙醇对GlyR的影响的多肽。该项目的长期目标是通过识别高度选择性的小肽来模拟或拮抗乙醇对单一靶标的影响，从而分离出单个假定的乙醇靶标的贡献。模拟乙醇在单个靶点上作用的多肽将允许研究人员仅模拟乙醇对该靶点的应用。相反，高特异性乙醇拮抗剂和乙醇的联合应用将模拟乙醇对除所选目标之外的每个目标的影响。从本质上讲，乙醇对单个目标的影响可以被分离出来，而不会干扰整个系统。通过这种方式，可以确定单个靶标在乙醇效应表达中所起的作用。了解个体靶标对酒精消费、成瘾和行为影响的重要性，将有助于合理、有针对性地开发与酒精中毒作斗争的药物。

DESCRIPTION (provided by applicant): Ethanol is a wildly used drug of abuse with a complex and poorly understood mechanism of action. A major hurdle in understanding the mechanism of alcohol action is the large number of putative ethanol targets. While many cellular components are known to be effected by alcohol, the contributions of each target to the overall physiological and behavioral effects of alcohol are unknown. We propose a method for isolating the effects of ethanol at a single target without altering the wild-type system. We plan to utilize phage display technology to identify peptides that bind with high specificity at a single putative ethanol target, the glycine receptor (GlyR). These peptides will then be tested electrophysiologically for modulatory effects on the GlyR. The goal of the proposed work is to identify peptides that either mimic or antagonize the effects of ethanol on the GlyR without affecting other ethanol targets. The long term objective of this project is to isolate the contribution of individual putative ethanol targets by identifying highly selective small peptides that either mimic or antagonize the effects of ethanol at a single target. Peptides that mimic ethanol actions at a single target would allow researchers to simulate the application of ethanol to only that target. Conversely, co-application of a highly specific ethanol antagonist and ethanol would simulate the effects of ethanol on every target except the selected one. Essentially, the effects of ethanol on a single target could be isolated without disturbing the overall system. In this way, the role of the individual target plays in the expression of ethanol's effects could be identified. Understanding the importance of individual targets to ethanol consumption, addiction and behavioral effects will allow for the rational, targeted development of drugs to combat alcoholism.