Genetic Basis of Opioid Reward and Aversion in Mice

7.项目摘要/摘要 处方阿片类药物的非医疗滥用是美国的一个主要社会经济问题。药效 已知虐待有遗传因素，流行病学研究表明，报告 快乐的经历最有可能转变为吸毒。因此，本申请旨在 确定阿片类药物奖赏和厌恶的遗传基础。我们将使用一个具有遗传信息的小鼠面板 品系和一种行为模型--位置条件反射试验--来筛选奖励的差异 对常见滥用处方阿片激动剂羟考酮(Oxy)的反应和对 阿片类拮抗剂纳洛酮(NAL)。在目标1中，我们将确定负责 氧奖赏和NAL厌恶的差异。在目标2中，我将接受信使核糖核酸测序(rna-seq)的培训。 鉴定这些基因组区域中差异表达的基因和剪接变异体可能导致 氧奖赏和NAL厌恶的变化。在目标3中，我们将产生携带小部分同源基因的小鼠 这些基因组区域缩小了潜在致病基因的名单。在这些地区中，我们将选择 通过分子遗传学技术进行进一步分析的差异表达的候选基因。这个 这项提案的结果将有助于识别与阿片类药物奖赏和厌恶有关的基因，这些基因将 对理解和治疗阿片类药物滥用的影响。 我的导师亚伯拉罕·帕尔默博士在这方面有几年的经验 项目，包括QTL定位、转录组分析和大规模的小鼠表型研究。Dr。 帕尔默在转录组分析方面的专业经验涉及到微阵列；因此，我聘请了Dr。 Jonathan Pritchard(共同导师)和Yoav Gilad博士(咨询委员会成员)提供导师和 关于RNA-seq的分析和技术方面的培训。此外，我还成立了一个咨询委员会 由Palmer博士、Pritchard博士、Gilad博士、Harriet de Wit博士和James Zacny博士组成。德维特博士有 在关于药物滥用的啮齿动物和人类研究方面具有广泛的背景和专业知识，尤其是 对拟议研究的翻译方面感兴趣。De Wit博士进行基因关联研究 涉及对滥用药物的敏感性，特别是与药物奖励等主观表型有关。因此，德博士 WIT处于有利地位，可以提供有关行为、遗传和药物方面的专家建议 建议，并可以提供应用结果的翻译上下文。扎克尼医生是阿片类药物方面的专家 药理学和人类阿片类药物特征的表型，并发表了多篇关于变异性的论文 对阿片类药物的生理和主观反应--特别是Oxy。扎克尼博士有兴趣申请 这一建议的结果被转译为关于人类阿片类药物敏感性的遗传基础的研究。 支持和补充这一提议的体制环境非常好。论 遗传和基因组项目结束时，在人文系有几个优秀的教员 与我互动的遗传学，包括我的委员会成员。我们有几个附属的研讨会系列 与我将参与的部门合作，包括进行中工作、每月研讨会系列、杂志俱乐部、 和模式生物遗传学。我的RNA-seq将至少有六台Illumina GA2机器 待检测的样本，其中三个位于加州大学校园。在神经科学方面，保罗博士 Vezina是NIDA资助的T32培训项目的负责人，该项目名为《药物滥用的神经精神药理学培训》 De Wit博士和Palmer博士都参加了 在这个项目中，S和其他几名神经科学教职员工都对 成瘾生物学。 我的长期目标是在一个主要研究领域建立自己的学术研究项目 大学旨在识别与药物滥用相关的表型相关基因，从 如本建议书所述放置条件化试验。在当前的QTL定位时代，转录组 分析现在是寻找导致复杂表型的基因的主要补充工具，例如 对药物敏感。因此，我在RNA-seq分析方面所接受的培训将帮助我实现这个职业目标。 在成瘾生物学领域，关于地点的遗传决定因素，存在着巨大的差距。 偏好。由于这种测试方法在成瘾生物学中的应用如此广泛，这一领域的研究无疑将 有助于我们理解现有的大量研究，并提供对 药物奖励和滥用的机制。我的实验室未来的提案将针对测试基因 在这项研究中确定了对与药物滥用有关的其他行为的多效性影响，包括 退出、自治、灭绝、重新巩固和恢复。我还对 奖励/厌恶和痛苦通路之间的交集，并希望进一步探索这一领域的研究 作为未来的提议。

7. Project Summary/Abstract The nonmedical abuse of prescription opioids is a major socioeconomic problem in the United States. Drug abuse is known to have a genetic component and epidemiological studies indicate that individuals reporting a pleasurable experience are most likely to transition to drug abuse. Accordingly, this application seeks to identify the genetic basis of opioid reward and aversion. We will use a genetically informative panel of mouse strains and a behavioral model- the place conditioning assay- to screen for differences in the rewarding response to the commonly abused prescription opioid agonist oxycodone (OXY) and the aversive response to the opioid antagonist naloxone (NAL). In Aim 1, we will identify genomic regions that are responsible for differences in OXY reward and NAL aversion. In Aim 2, I will receive training in mRNA sequencing (RNA-seq) to identify differentially expressed genes and splice variants in these genomic regions as potentially causal for variation in OXY reward and NAL aversion. In Aim 3, we will generate congenic mice carrying small portions of these genomic regions to narrow the list of potential genes responsible. Within these regions, we will choose candidate genes that are differentially expressed for further analysis via molecular genetics techniques. The outcome of this proposal will aid in identifying genes contributing to opioid reward and aversion which will have implications for understanding and treating opioid abuse. My mentor, Dr. Abraham Palmer, has several years of experience with regard to all aspects of this project, including QTL mapping, transcriptome analysis, and large-scale mouse phenotyping studies. Dr. Palmer's history of expertise for transcriptome analysis has involved microarrays; thus, I have recruited Dr. Jonathan Pritchard (Co-Mentor) and Dr. Yoav Gilad (advisory committee member) to provide mentorship and training in the analytical and technical aspects of RNA-seq. Additionally, I have formed an advisory committee consisting of Dr. Palmer, Dr. Pritchard, Dr. Gilad, Dr. Harriet de Wit, and Dr. James Zacny. Dr. de Wit has extensive background and expertise in both rodent and human studies of drug abuse and is particularly interested in the translational aspect of the proposed research. Dr. de Wit conducts genetic association studies involving sensitivity to drugs of abuse, notably with subjective phenotypes such as drug reward. Thus, Dr. de Wit is well-positioned to provide expert advice on the behavioral, genetic, and pharmacological aspects of this proposal and can offer a translational context from which to apply the results. Dr. Zacny is an expert in opioid pharmacology and human phenotyping of opioid traits and has published multiple papers regarding variability in the physiological and subjective response to opioids- in particular OXY. Dr. Zacny is interested in applying the results of this proposal to translational studies regarding the genetic basis of opioid sensitivity in humans. The institutional environment for supporting and complementing this proposal is excellent. On the genetic and genomic end of the project, there are several excellent faculty in the Department of Human Genetics with whom I interact, including members of my committee. We have several seminar series affiliated with the department that I will participate in, including Work in Progress, Monthly Seminar Series, Journal Club, and Genetics of Model Organisms. There will be at least six Illumina GA2 machines in place for my RNA-seq samples to be run, three of which are located on the U of C campus. On the neuroscience end, Dr. Paul Vezina heads a NIDA-funded T32 training program, "Neuropsychopharmacology Training for Drug abuse Research" and seminar series that meets twice per month. Dr. de Wit and Dr. Palmer are both participating P.I.'s in this program, along with several other neuroscience faculty members who are all interested in addiction biology. My long-term objective is to establish my own academic research program at a major research university aimed at identifying genes involved with phenotypes associated with drug abuse, starting with the place conditioning assay as outlined in this proposal. In the current era of QTL mapping, transcriptome analysis is now a mainstay complementary tool for finding genes contributing to complex phenotypes such as drug sensitivity. Thus, the training I receive in RNA-seq analysis will help me to realize this career goal. There is a huge gap in the field of addiction biology regarding the genetic determinants of place preference. Because this assay is so widely used in addiction biology, this area of research will undoubtedly contribute to our understanding of the multitude of existing studies and provide novel insight into the mechanisms of drug reward and abuse. Future proposals from my laboratory will be aimed at testing genes identified in this study for pleiotropic effects on other behaviors associated with drug abuse, including withdrawal, self-administration, extinction, reconsolidation, and reinstatement. I am also interested in the intersection between reward/aversion and pain pathways and would like to further explore this area of research as a future proposal.