Genetics of Cocaine Sensitivity in Drosophila

摘要 非法使用可卡因和其他毒品是一个世界性的健康问题。美国国家药物滥用研究所 估计因使用烟草、酒精和非法药物而造成的药物滥用和成瘾的总成本为820美元 每年数十亿美元，使药物滥用成为美国代价最高的公共卫生问题。仅非法使用毒品一项 在医疗保健、生产力损失、犯罪、监禁和毒品执法方面占1930亿美元。在……里面 人类对可卡因和其他药物的易感性有很强的遗传成分，但很少 在识别潜在的变异和基因方面取得了进展，部分原因是很难获得 由于药物滥用被定为刑事犯罪，样本量足够大；药物暴露的变化， 包括同时接触多种药物、酒精和尼古丁；与其他 神经精神障碍。使用果蝇等模式生物可以缓解这些问题 黑猩猩。除了饲养成本低、体型小、世代间隔短等优点外，果蝇 拥有丰富的公共可用的遗传资源。重要的是，精神刺激剂对人的许多影响 是在果蝇身上复制的。大约67%的苍蝇基因与人类同源，因此获得了洞察力 来自果蝇的基因具有翻译潜力。在过去的支持期间，我们使用了D。 具有全基因组测序的近交系野生来源苍蝇品系的黑腹果蝇遗传参考组，以及 从DGRP系衍生的异交高级杂交群体(AIP)，进行全基因组关联 (GWA)药物消费行为和基因表达的分析。这些分析表明，变种 与药物消费相关的表型主要位于非编码区，并且 推测是通过基因调控发挥表型效应。我们推导出了基因调控 来自自然发生的基因表达遗传变异的网络，并构建了基因图谱 果蝇大脑在单细胞分辨率下暴露可卡因后的表达变化。挑战 现在要了解的是变异如何共同作用来影响药物偏好的变异，并确定 使用系统遗传学分析的潜在分子网络。在这里，我们建议使用1200个新的DGRP 用于定位与可卡因偏好相关的自然发生的变异和基因的品系大大增加 比我们以前的研究更强大和准确，执行系统的基因分析来推断因果调控 与可卡因偏好相关的网络，并使用生殖系基因编辑来证明遗传的因果关系 与可卡因偏好和基因调控网络的关联。从这些研究中获得的信息 可以作为随后在哺乳动物系统和人类群体中进行翻译研究的蓝图 基于正交学和基本生物过程的进化守恒，并扩展了 与人类药物敏感性变异相关的遗传框架超出候选药物的狭窄范围 到目前为止已经检查过的基因。

SUMMARY Illegal use of cocaine and other drugs is a worldwide health problem. The National Institute on Drug Abuse estimates the total costs of drug abuse and addiction due to use of tobacco, alcohol and illegal drugs at $820 billion a year, making substance abuse the most costly public health problem in the nation. Illicit drug use alone accounts for $193 billion in health care, productivity loss, crime, incarceration, and drug enforcement. In humans, susceptibility to the effects of cocaine and other drugs has a strong genetic component, but little progress has been made in identifying the underlying variants and genes, in part due to difficulty in obtaining sufficiently large sample sizes because of criminalization of substance abuse; variation in drug exposure, including simultaneous exposure to multiple drugs, alcohol and nicotine; and comorbidity with other neuropsychiatric disorders. These problems can be mitigated using model organisms, such as Drosophila melanogaster. In addition to benefits of low rearing costs, small size and a short generation interval, Drosophila has a wealth of publically available genetic resources. Importantly, many effects of psychostimulants on people are replicated in flies. Approximately 67% of fly genes have human orthologs, and therefore insights gained from Drosophila have translational potential. During the past period of support, we have used the D. melanogaster Genetic Reference Panel of inbred wild-derived fly strains with fully sequenced genomes, and outbred advanced intercross populations (AIPs) derived from DGRP lines, to perform genome wide association (GWA) analyses of drug consumption behaviors and gene expression. These analyses showed that variants associated with drug consumption phenotypes were largely located in non-coding genomic regions, and presumably exert their phenotypic effects via modulation of gene regulation. We derived gene regulatory networks from naturally occurring genetic variation in gene expression and constructed an atlas of gene expression changes in the Drosophila brain following cocaine exposure at single cell resolution. The challenge now is to understand how variants act jointly to affect variation in drug preference, and to determine the underlying molecular networks using systems genetics analyses. Here, we propose to use 1200 new DGRP lines to map naturally occurring variants and genes associated with cocaine preference with greatly increased power and precision than our previous studies, perform a systems genetic analysis to infer causal regulatory networks associated with cocaine preference, and use germline gene editing to prove causality of the genetic associations with cocaine preference and gene regulatory networks. Information obtained from these studies can serve as a blueprint for subsequent translational studies in mammalian systems and human populations based on orthology and evolutionary conservation of fundamental biological processes, and expand the genetic framework associated with variation in human drug susceptibility beyond the narrow range of candidate genes examined to date.