Neuromolecular Drivers of Impulsivity in Addictive Disorders

项目摘要/摘要 本提案要求支持一项全面的培训计划，使Noelle C.Anastasio博士、 拓宽、提高和完善独立研究成果所需的技术技能 职业生涯。我的长期职业目标是在一家研究密集型学院获得终身教职。 机构和建立一个研究计划，专注于最终的科学目标，以理解 精神药理学的冲动性特征，以促进成瘾的预防和治疗。各方面 药物滥用，包括开始吸毒，从随意使用药物过渡到强制使用药物， 维持寻求毒品的行为以及恢复寻求毒品的倾向与 天生的冲动程度。脑内5-羟色胺2C受体(5-HT2CR)功能失调 中皮质边缘环路参与了成瘾周期的这些阶段，以及 冲动表型的表现。有证据表明，5-羟色胺(5-羟色胺)神经传递 通过其同源5-HT2C受体(5-HT2CR)和谷氨酸通过亲离子神经传递 谷氨酸N-甲基-D-天冬氨酸受体(NMDAR)在认知和/或行为中起关键作用 冲动和成瘾行为的维度，特别是在mPFC中，这是决策的一个组成部分 和目标导向的行为。因此，mPFC中5-羟色胺和谷氨酸系统之间的相互作用可能 导致可卡因成瘾所致的认知障碍。我将在培训期间接受多方面的培训。 K99指导阶段(1-2年；目标1-2)，由协作导师团队提供，包括 用于分析的分子技术(例如，蛋白质交联、细胞系的建立、生物反应分析) 受体的亚细胞定位和信号转导调节，以及复杂的行为 神经科学/药理学/分子遗传学技术(例如，免疫组织化学、颅内 微输注、病毒介导的基因转移)。我还将接受职业发展方面的指导， 资质、实验室工作人员和学生管理以及财政和规划责任对于 经营一个独立的实验室。在R00独立阶段提出的研究计划 (3-5年；AIMS 3-4)在此培训的基础上，更具体地关注mPFC的神经生物学 NMDAR和5-HT2CR：NMDAR作为功能变阻器在固有基因表达中的作用 冲动与可卡因相关的复发事件。中心假设是5-HT2CR和 NMDAR功能驱动mPFC输出，并且这种平衡的损失有助于固有的冲动性， 导致容易发展成瘾障碍和相关的复发事件。我们将测试 这一假说和揭示了导致固有冲动的神经分子驱动因素以及可卡因- 与可卡因相关的复吸事件相关的诱发冲动(例如，线索反应性)。要解决这个问题 假设，已经制定了四个具体目标：目标1)建立mPFC 5-HT2CR在固有的 冲动性；目的2)确定mPFC 5-HT2CR在聚合冲动性/线索反应性中的作用；目的3)阐明 MPFC NMDAR在内在冲动和聚合冲动/线索反应性中的作用；目标4)探索 5-HT2CR：NMDAR复合体的体外生物学研究我们将确定神经元5-HT2CR和 NMDAR系统管理冲动，重新平衡这些系统最终可能支持行为 在以冲动为特征的紊乱中恢复。总之，这些创新的翻译研究将是第一个 系统探讨5-HT2CR和NMDAR介导的功能在成瘾中的作用和相互作用。 相关表型，最终允许设计有针对性的药物治疗以促进 戒除和防止成瘾障碍的复发。

PROJECT SUMMARY/ABSTRACT This proposal requests support for a comprehensive training plan that will enable Noelle C. Anastasio, Ph.D., to broaden, enhance, and refine technical skills that are necessary for a productive independent research career. My long-term career goal is to obtain a tenure-track faculty position at a research-intensive academic institution and build a research program focused upon the ultimate scientific goal to understand the psychopharmacology of the impulsivity trait to advance prevention and treatment of addiction. Various aspects of drug abuse, including the initiation of drug-taking, the transition from casual to compulsive drug use, the maintenance of drug-seeking behaviors as well as the penchant to reinstate drug-seeking correlate with high levels of inherent impulsivity. Dysregulation in serotonin (5-HT) 2C receptor (5-HT2CR) function within the mesocorticolimbic circuit has been implicated in these stages of the addiction cycle as well as the manifestation of the impulsive phenotype. There is evidence that both serotonin (5-HT) neurotransmission through its cognate 5-HT2C receptor (5-HT2CR) and glutamate neurotransmission through the ionotropic glutamate N-methyl-D-aspartate receptor (NMDAR) play key roles in the cognitive and/or behavioral dimensions of impulsivity and addictive behaviors, especially in the mPFC, a region integral to decision-making and goal-directed behavior. Thus, the interaction between 5-HT and glutamate systems in the mPFC may contribute to the cognitive impairments seen in cocaine addiction. I will receive multifaceted training during the K99 mentored phase (Years 1-2; Aims 1-2) from a team of collaborating mentors that includes training in molecular techniques (e.g., protein crosslinking, creation of cell lines, bioresponsive assays) to analyze receptor subcellular localization and signal transduction regulation, as well as sophisticated behavioral neuroscience/pharmacology/molecular genetics techniques (e.g., immunohistochemistry, intracranial microinfusion, viral-mediated gene transfer). I will also receive mentoring in career development, grantsmanship, laboratory staff and student management and fiscal and planning responsibilities critical to running an independent laboratory. The research plan that is proposed during the R00 independent phase (Years 3-5; Aims 3-4) builds on this training to focus more specifically on the neurobiology of the mPFC NMDAR and the 5-HT2CR:NMDAR heteromeric complex as functional rheostats in expression of inherent impulsivity and cocaine-associated relapse events. The central hypothesis is that a balance of 5-HT2CR and NMDAR function drives mPFC output and that the loss of this balance contributes to inherent impulsivity, leading to vulnerability for the development of addictive disorders and associated relapse events. We will test this hypothesis and uncover neuromolecular drivers responsible for inherent impulsivity as well as the cocaine- evoked impulsivity in relation to cocaine-associated relapse events (e.g., cue reactivity). To address this hypothesis, four specific aims have been formulated: Aim 1) establish the role for mPFC 5-HT2CR in inherent impulsivity; Aim 2) establish the role for mPFC 5-HT2CR in aggregate impulsivity/cue reactivity; Aim 3) elucidate the role for mPFC NMDAR in inherent impulsivity and aggregate impulsivity/cue reactivity; Aim 4) explore the biology of the 5-HT2CR:NMDAR complex in vitro and ex vivo. We will determine that neuronal 5-HT2CR and NMDAR systems govern impulsivity and that rebalancing these systems may ultimately support behavioral recovery in disorders marked by impulsivity. Together, these innovative translational studies will be the first to systemically explore the contribution and interaction of 5-HT2CR- and NMDAR-mediated function in addiction- relevant phenotypes, ultimately allowing for the design of targeted pharmacotherapeutics to promote abstinence and prevent relapse in addictive disorders.