Genetics of Cocaine Sensitivity in Drosophila

总结 非法使用可卡因和其他毒品是一个世界性的健康问题。国家药物滥用研究所 估计由于使用烟草、酒精和非法药物而导致的药物滥用和成瘾的总成本为820美元 这使得药物滥用成为美国最昂贵的公共卫生问题。仅非法药物使用 占1930亿美元的医疗保健，生产力损失，犯罪，监禁和禁毒执法。在 在人类中，对可卡因和其他药物影响的易感性具有很强的遗传成分，但很少 在确定潜在的变异和基因方面取得了进展，部分原因是难以获得 由于将药物滥用定为刑事犯罪，样本量足够大;药物接触的变化， 包括同时暴露于多种药物、酒精和尼古丁;以及与其他 神经精神障碍这些问题可以通过使用模式生物来缓解，例如果蝇。 黑腹菌除了饲养成本低、体型小和世代间隔短等优点外， 有着丰富的生物遗传资源。重要的是，精神兴奋剂对人的许多影响 在苍蝇中复制。大约67%的果蝇基因具有人类直系同源物， 都有翻译潜能。在过去的支持期间，我们使用了D。 具有完全测序的基因组的近交野生来源的果蝇品系的黑腹果蝇遗传参考组，和 从DGRP系衍生的远交高级互交群体（AIP），以进行全基因组关联 (GWA)药物消费行为和基因表达的分析。这些分析表明， 与药物消耗表型相关的基因主要位于非编码基因组区域， 可能通过基因调控的调节发挥其表型效应。我们推导出了基因调控 从基因表达中自然发生的遗传变异中构建了一个基因图谱， 在单细胞分辨率下，可卡因暴露后果蝇脑中的表达变化。的挑战 现在是了解变异如何共同作用，影响药物偏好的变化，并确定 使用系统遗传学分析潜在的分子网络。在这里，我们建议使用1200个新DGRP 线映射自然发生的变异和基因与可卡因的偏好大大增加 的能力和精确度，进行系统的遗传分析，以推断因果调节 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系。从这些研究中获得的信息 可以作为哺乳动物系统和人类群体中后续转化研究的蓝图 基于基本生物过程的正形学和进化保护， 与人类药物敏感性变异相关的遗传框架超出了候选药物的狭窄范围 基因检测到目前为止

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.