Molecular Studies of Cocaine Action in Brain

项目负责人/主要研究者（最后，第一，中间）：Nestler，Eric，J. 项目摘要 该R 01补助金旨在更好地了解慢性暴露于 滥用药物会引起大脑奖赏回路的长期变化， 成瘾状态下的行为异常我们的工作集中在转录因子AFosB， 这是在丘脑核（NAc）和其他关键的大脑奖励区域中诱导的， 几乎所有滥用药物的管理。AFosB的独特之处在于，它只能累积到可观的水平 并且由于其不寻常的稳定性，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发生长期变化的机制之一。 导致成瘾的表达。事实上，大量证据支持这样的观点：诱导 NAc中的AFosB介导了一种高度奖励和动机的状态，这可能有助于 成瘾过程在这次要求的R37更新中，我们将描述通过哪些确切的机制， AFosB采取了这些行动。使用最先进的全基因组染色质分析，我们将确定基因， 在可卡因和阿片类药物自我给药的情况下，NAc是AFosB的直接靶点。这样的基因， 反过来，提供了新的见解药物诱导的神经和行为的分子和细胞基础， 可塑性。有趣的是，当AFosB与其靶基因结合时，它可以激活或抑制它们。我们 一种假说认为，这种激活与抑制作用是由受影响的细胞的染色质环境决定的。 基因此外，NAc中部分非重叠基因受AFosB调节，以响应两种药物。 尽管这可能部分是由药物诱导的部分不同的神经元亚群介导的， AFosB，我们再次假设这种特异性也部分由其他染色质的差异介导 可卡因和阿片类药物在特定基因上引起的变化。除了探讨AFosB对靶细胞的调节作用外， 基因，我们还将描述几种机制，这是至关重要的，在确定的数量和活动， AFosB在NAc中诱导。总之，这些研究将确定滥用药物的许多新行为， 可以用来开发更好的诊断测试和治疗成瘾。 PHS 398/2590（Rev. 06/09）第2页 项目负责人/主要研究者（最后一名、第一名、中间名）：Nestler，EriC，J. 2004年至2005年初步预算期间的详细预算 仅直接费用16年4月1日17年3月31日 人员名单（仅适用于申请组织） 使用Cal、Acad或Summer输入投入项目的月份 输入请求的薪资和附带福利的美元金额（省略美分） 加州大学的作用夏季研究所基本工资福利 名称项目数 Eric Nestler PD/PI 1.2 181，500 18，150 5，082 23，232 李申教授1.2 103，378 0 0 0 Deena步行者博士后6 49，680 24，840 6，955 31，795 Jaclyn Rabkin格拉德研究生12 33，500 33，500 0 33，500 Erin Calipari博士后6 48，000 24，000 6，720 30，720 Dominika Burek资源助理12 35，000 35，000 9，800 44，800 小计- 咨询人费用 设备（逐项列出） 用品（按类别逐项列出） 一般用品-94 510 动物费用-40 490 ChIP-seq和RNA-seq约18，000 旅行 神经科学学会年会 住院护理费用 门诊护理费用 改建和翻修（按类别逐项列出） 其他费用（按类别分列） 出版物-5 000 生物信息服务-25，000 格拉德研究生学费-3 591人;健康保险-4 909人 联合体/采购成本 初步预算期间直接费用小计 联合体/采购成本 初步预算期间直接费用共计 PHS 398（2012年8月8日至2015年8月31日批准修订版） 第3页_ ·135，490 28，557 164，047 153,000 3,000 38,500 直接成本 （/<em 7a，Face Page; 358，547美元 设施和行政费用 三十五万八千五百四十七美元 OMB编号0925-0001 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