Molecular Mechanisms of Susceptibility to Drug Use

项目概要/摘要 从吸毒到滥用并最终到依赖的转变可能是由生物因素介导的 在使用药物之前就存在。尽管众所周知，长期接触滥用药物会扰乱许多 信号通路，但人们对介导成瘾易感性的分子机制知之甚少。那里 对识别成瘾易感性的早期生物标志物以改善成瘾非常感兴趣 预防策略。大多数研究都是在物质依赖个体中进行的，其中 “敏感性”和“后果”之间的分离是模糊的。我最近发现了一种行为 大鼠的表型能够可靠地预测未来的吸毒行为，并确定了三种潜在的蛋白质 成瘾易感性介质：分选连接蛋白 1 (SNX1)、兰尼碱受体 2 (RYR2) 和 ataxin 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，并且之前已被认为与成瘾有关。这些表达的差异究竟如何 蛋白质影响成瘾易感性背后的信号级联尚不清楚。总体目标 该提案的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如何与遗传和/或环境有机械联系 成瘾风险的组成部分。为了实现这一目标，拟议的研究将结合复杂的 使用病毒、蛋白质组和生物信息学方法进行行为和计算评估。在目标 1 中我会 使用可诱导和可逆的病毒构建体双向操纵 SNX1、RYR2 和 ATXN2L 并确定 SNX1、RYR2 和 ATXN2L 表达的变化如何改变 甲基苯丙胺自我给药和蛋白质信号传导机制。在目标 2 中，我将确定变化是否 SNX1、RYR2 和 ATXN2L 的表达在遗传成瘾易感性模型中发生改变，并且 与成瘾风险增加有关。在目标 3 中，我将确定 SNX1、RYR2、 ATXN2L 在环境成瘾易感性模型中发生改变，并与增加 成瘾风险。完成这些目标将为成瘾的信号机制带来新的见解 易感性可以识别成瘾风险的早期生物标记并改进当前的策略 预防成瘾。首席研究员将接受病毒和病毒方面的指导和技术培训 细胞信号和细胞机制专家的蛋白质组技术以及病毒技术 动机行为。耶鲁大学和精神病学系提供卓越的设施和 完成拟议实验的资源，以及拥有卓越的声誉和跟踪 指导早期调查人员并将其转变为独立调查人员的记录。拟议的 培训、教育和研究将为 PI 提供技术和专业培训，使其成为一名 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.