Molecular Mechanisms of Susceptibility to Drug Use

项目概要/摘要 从药物使用到滥用并最终到依赖的转变可能是由生物因素介导的 在吸毒前就存在的虽然长期暴露于滥用药物是众所周知的扰乱许多 信号通路，对介导成瘾易感性的分子机制知之甚少。那里 对鉴定成瘾易感性的早期生物标志物以改善成瘾性是相当感兴趣的 预防战略。大多数研究都是在物质依赖者中进行的， “易感性”和“后果”之间的分离是模糊的。我最近发现了一种行为 在大鼠中的表型，可靠地预测未来的吸毒行为，并确定了三种蛋白质作为潜在的 成瘾易感性介质：分类连接蛋白1（SNX 1）、兰尼碱受体2（RYR 2）和共济失调蛋白2样 （ATXN2L）。这些蛋白质参与细胞内运输、钙信号传导和细胞骨架 重组，以前与成瘾有关。这些基因表达的差异 蛋白质影响成瘾易感性潜在的信号级联尚不清楚。总体目标 这项提议的一个重要目的是确定介导成瘾的蛋白质的功能和分子作用 易感性，并研究这些因素是如何与遗传和/或环境 成瘾风险的组成部分。为了实现这一目标，拟议的研究将联合收割机 使用病毒、蛋白质组学和生物信息学方法进行行为和计算评估。目标1我会 使用可诱导和可逆的病毒构建体双向操纵SNX 1、RYR 2和 ATXN 2L，并确定SNX 1，RYR 2和ATXN 2L表达的变化如何改变 甲基苯丙胺自我管理和蛋白质信号传导机制。在目标2中，我将确定变化是否 SNX 1、RYR 2和ATXN 2L的表达在遗传成瘾易感性模型中发生改变， 与成瘾风险增加有关。在目标3中，我将确定SNX 1，RYR 2， 和ATXN 2L在环境成瘾易感性模型中改变，并与增加的 成瘾风险这些目标的完成将产生对成瘾信号机制的新见解 易感性，可以识别成瘾风险的早期生物标志物，并改善目前的策略， 预防成瘾。主要研究者将接受病毒和 蛋白质组学技术由细胞信号传导和细胞机制方面的专家，以及病毒技术， 动机行为。耶鲁大学和精神病学系提供卓越的设施， 资源完成拟议的实验，以及具有特殊的声誉和跟踪 指导早期研究者并将其转变为独立研究者的记录。拟议 培训、教育和研究将为PI提供技术和专业培训，使其成为 成功的独立成瘾调查员

PROJECT SUMMARY / ABSTRACT The transition from drug use to abuse and, eventually, to dependence may be mediated by biological factors that are present prior to drug use. Although chronic exposure to drugs of abuse is known to disrupt many signaling pathways, little is known about the molecular mechanisms that mediate addiction susceptibility. There is considerable interest in identifying early biomarkers for addiction susceptibility to improve addiction prevention strategies. Most studies have been conducted in substance-dependent individuals where the dissociation between ‘susceptibility’ and ‘consequence’ is ambiguous. I have recently identified a behavioral phenotype in rats that reliably predicts future drug-taking behaviors and identified three proteins as potential mediators of addiction susceptibility: sorting nexin 1 (SNX1), ryanodine receptor 2 (RYR2), and ataxin 2-like (ATXN2L). These proteins are involved in intracellular trafficking, calcium signaling and cytoskeleton reorganization, and have been previously linked to addiction. Precisely how differences in expression of these proteins impact the signaling cascades underlying addiction susceptibility is not known. The overarching goal of this proposal is to determine the functional and molecular role of proteins that mediate addiction susceptibility and investigate how these factors are mechanistically linked to genetic and/or environmental components of risk for addiction. To accomplish this goal, the proposed research will combine sophisticated behavioral and computational assessments with viral, proteomic and bioinformatic approaches. In Aim 1 I will use an inducible and reversible viral construct to bi-directionally manipulate expression of SNX1, RYR2, and ATXN2L and also determine how changes in expression of SNX1, RYR2, and ATXN2L alter methamphetamine self-administration and protein signaling mechanisms. In Aim 2 I will determine if variation in the expression of SNX1, RYR2, and ATXN2L is altered in a model of genetic addiction susceptibility and is associated with increased addiction risk. In Aim 3 I will determine if variation in expression of SNX1, RYR2, and ATXN2L is altered in a model of environmental addiction susceptibility and is associated with increased addiction risk. Completion of these aims will generate new insights into the signaling mechanisms of addiction susceptibility that could identify early biological markers of risk for addiction and improve current strategies for addiction prevention. The Principal Investigator will receive mentorship and technical training in viral and proteomic technologies by experts in cell signaling and cellular mechanisms, and viral technologies in motivated behaviors. Yale University and the Department Psychiatry provide exceptional facilities and resources for completing the proposed experiments, as well as having an exceptional reputation and track record for mentoring and transitioning early-stage investigators in to independent investigators. The proposed training, education and research will provide the PI with the technical and professional training to become a successful, independent addiction investigator.