Molecular Studies of Cocaine Action in Brain

项目主任/首席调查员(最后、第一、中间)：Nestler，Eric，J. 项目摘要 这项R01拨款旨在更好地了解慢性接触的分子机制 滥用药物会导致大脑奖赏回路的长期变化，从而导致复杂的 定义成瘾状态的行为异常。我们的工作重点是转录因子AFosB， 它是在伏核(NAC)和其他关键的大脑奖赏区域诱导的，以响应慢性 管理几乎所有的滥用药物。AFosB的独特之处在于它只积累到可察觉的水平 在长期接触药物后，由于其不同寻常的稳定性，可持续数周至数月的戒断。 因此，AFosB是慢性药物暴露可以驱动基因长期变化的一种机制 导致上瘾的表情。事实上，相当多的证据支持这样一种观点，即诱导 NAC的AFosB调节一种高奖励和动机的状态，这可能有助于在某些方面 上瘾的过程。在这次请求的R37续订中，我们将描述通过以下方式实现的精确机制 AFosB执行这些操作。使用最先进的全基因组染色质分析，我们将在 在可卡因和鸦片类药物自我给药的背景下，NAC是AFosB的直接目标。这样的基因， 反过来，为药物诱导的神经和行为的分子和细胞基础提供了新的见解 可塑性。有趣的是，当AFosB与其目标基因结合时，它可以激活或抑制它们。我们的 假说是，这种激活和抑制作用是由受影响的染色质环境决定的。 吉恩。此外，在NAC中，部分不重叠的基因受到AFosB的调控，对这两种药物做出反应。 尽管这可能部分是通过药物诱导的部分不同的神经元亚群来调节的 AFosB，我们再次假设这种特异性也部分地通过其他染色质的差异来调节 可卡因和鸦片类药物在特定基因上引起的变化。除了探索AFosB对靶标的调控 基因，我们还将描述几种在决定酶的数量和活性方面至关重要的机制。 在NAC中诱导出AFosB。总而言之，这些研究将确定许多新的滥用药物行为， 可用于开发改进的成瘾诊断测试和治疗方法。 PHS 398/2590(06/09版)第2页续格式页 方案主任/首席调查员(最后、第一、中间)：Nestler，Eric，J. 初始预算期间的详细预算，从 直接成本仅4/1/16 3/31/17 人员列表(仅适用于申请组织) 使用Cal、ACAD或Summer输入用于项目的月份 输入申请薪资和附带福利的申请金额(省略美分) 在Cal上的角色。阿卡德。暑期班基本工资福利 名称项目月薪申请福利合计 Eric Nestler PD/PI 1.2 181,500 18,150 5,082 23,232 李申学院1.2 103,378 0 0 蒂娜·沃克博士后6 49,680 24,840 6,955 31,795 Jaclyn Rabkin研究生12 33,500 33,500 0 33,500 Erin Calipari博士后6 48,000 24,000 6,720 30,720 Dominika Britk Res.Asst 12 35,000 35,000 9,800 44,800 小计- 顾问费 设备(分项) 用品(按类别分项列出) 一般用品--94,510 动物成本-40,490 ChlP-seq和RNA-seq-18,000 旅行 神经科学学会年会 住院护理费用 门诊费用 改建和翻新(按类别分列) 其他费用(按类别分项列出) 出版物--5,000份 生物信息服务--25,000 研究生学费-3,591；医疗保险-4,909 联合体/合同成本 初步预算期的直接费用小计 联合体/合同成本 初始预算期的总直接成本 小灵通398(截至2015年8月31日批准08/12修订版) 第3页_ ·135,490 28,557 164,047 153,000 三千 38,500人 直接成本 (/&lt；em 7a，正面页；358,547美元 设施和行政费用 358,547美元 管理及预算局编号0925-0001 表单第4页

Program Director/Principal Investigator (Last, First, Middle): Nestler, Eric, J. Project Summary This R01 grant is aimed at better understanding the molecular mechanisms by which chronic exposure to drugs of abuse induces long-lasting changes in the brain's reward circuits that contribute to the complex behavioral abnormalities that define an addicted state. Our work focuses on the transcription factor, AFosB, which is induced in the nucleus accumbens (NAc) and other key brain reward regions in response to chronic administration of virtually all drugs of abuse. AFosB is unique in that it accumulates to appreciable levels only after chronic drug exposure and, because of its unusual stability, persists for weeks-months of withdrawal. AFosB is thus one mechanism by which chronic drug exposure can drive long-lasting changes in gene expression that contribute to addiction. Indeed, considerable evidence supports the view that induction of AFosB in NAc mediates a state of heightened reward and motivation that could contribute to aspects of the addiction process. In this requested R37 renewal, we will characterize the precise mechanisms through which AFosB exerts these actions. Using state-of-the-art genome-wide chromatin assays, we will identify the genes in the NAc that are direct targets for AFosB in the context of cocaine and opiate self-administration. Such genes, in turn, provide novel insight into the molecular and cellular basis of drug-induced neural and behavioral plasticity. Interestingly, when AFosB binds to its target genes, it can either activate or repress them. Our hypothesis is that such activation vs. repressive actions are determined by the chromatin milieu of the affected gene. As well, partially non-overlapping genes are regulated by AFosB in NAc in response to the two drugs. Although this is likely mediated in part by the partly distinct subsets of neurons in which the drugs induce AFosB, we once again hypothesize that this specificity is also mediated partly by differences in other chromatin changes that cocaine and opiates induce at specific genes. In addition to exploring AFosB's regulation of target genes, we will also characterize several mechanisms that are crucial in determining the amount and activity of AFosB induced in the NAc. Together, these studies will identify many novel actions of drugs of abuse which can be exploited for the development of improved diagnostic tests and treatments for addiction. PHS 398/2590 (Rev. 06/09) Page 2 Continuation Format Page ProgramDirector/PrincipalInvestigator(Last,First,Middle): Nestler,EriC,J. DETAILED BUDGET FOR INITIAL BUDGET PERIOD FROM THROUGH DIRECT COSTS ONLY 4/1/16 3/31/17 List PERSONNEL (Applicant organization only) Use Cal, Acad, or Summer to Enter Months Devoted to Project Enter Dollar Amounts Requested (omit cents) for Salary Requested and Fringe Benefits ROLE ON Cal. Acad. Summer INST.BASE SALARY FRINGE NAME PROJECT Mnths Mnths Mnths SALARY REQUESTED BENEFITS TOTAL Eric Nestler PD/PI 1.2 181,500 18,150 5,082 23,232 Li Shen Faculty 1.2 103,378 0 0 0 Deena Walker Postdoc 6 49,680 24,840 6,955 31,795 Jaclyn Rabkin Grad Student 12 33,500 33,500 0 33,500 Erin Calipari Postdoc 6 48,000 24,000 6,720 30,720 Dominika Burek Res. Asst 12 35,000 35,000 9,800 44,800 SUBTOTALS - CONSULTANT COSTS EQUIPMENT (Itemize) SUPPLIES (Itemize by category) General Supplies - 94,510 Animal Costs - 40,490 ChlP-seq and RNA-seq -18,000 TRAVEL Society for Neuroscience annual meeting INPATIENT CARE COSTS OUTPATIENT CARE COSTS ALTERATIONS AND RENOVATIONS (Itemize by category) OTHER EXPENSES (Itemize by category) Publication - 5,000 Bioinformatic Services - 25,000 Grad Student Tuition - 3,591; Health Insurance - 4,909 CONSORTIUM/CONTRACTUAL COSTS SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD CONSORTIUM/CONTRACTUAL COSTS TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD PHS 398 (Rev. 08/12 Approved Through 8/31/2015) Page 3_ · 135,490 28,557 164,047 153,000 3,000 38,500 DIRECT COSTS (/<em 7a, Face Page; $ 358,547 FACILITIES AND ADMINISTRATIVE COSTS $ 358,547 OMB No. 0925-0001 Form Page 4