Systems Genetics of Cocaine Preference in Drosophila

果蝇可卡因偏好的系统遗传学

• 批准号: 10675195
• 负责人: Jeffrey S Hatfield
• 金额: $ 4.77万
• 依托单位: CLEMSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-08 至 2025-09-07
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10675195
• 关键词: Acute; Affect; Alleles; Animal Model; Animals; Behavioral; Behavioral Assay; Binding; Biological Assay; Blood capillaries; Brain; Candidate Disease Gene; Chromosome Mapping; Cocaine; Cocaine use disorder; Collection; Consumption; Data; Development; Drosophila genus; Drosophila melanogaster; Emergency department visit; Environment; Environmental Risk Factor; Exhibits; Genes; Genetic; Genetic Models; Genetic Techniques; Genetic study; Goals; Grooming; Haplotypes; Health; Heritability; Human; Human Genome; Hyperactivity; Inbreeding; Libraries; Locomotion; Maps; Measures; Mediating; Methods; Ontology; Pathway interactions; Patient Recruitments; Pharmaceutical Preparations; Phenotype; Population; Public Health; RNA; RNA Interference; Sample Size; Sampling; Seizures; Self Administration; Structure; Substance Use Disorder; System; Testing; Training; Twin Studies; United States; Variant; advanced system; behavior test; candidate validation; career; cocaine exposure; cocaine overdose; cocaine self-administration; cocaine use; cost; cost effective; dopamine transporter; follow-up; gene discovery; genetic analysis; genetic approach; genetic variant; genome wide association study; health economics; knock-down; model organism; novel; preference; screening; sexual dimorphism; socioeconomics; trait; transcriptome sequencing; transcriptomics; translational potential; validation studies; whole genome

PROJECT SUMMARY Cocaine Use Disorder (CUD) represents a significant public health and socioeconomic concern. Cocaine is involved in 40% of drug-related emergency room visits, and rates of cocaine overdose in the United States are climbing steadily. Yet, the genetic underpinnings that predispose to CUD remain poorly understood. Twin studies on the genetics of CUD provide heritability estimates between 42-79%. However, significant heritability estimates do not indicate which genes are involved, and how they might be functionally relevant. Genome-wide association studies (GWAS) of CUD are limited by inability to control environmental factors; the haplotype structure of the human genome limiting mapping precision; and small sample sizes due to difficulty recruiting participants because of criminalization of cocaine use. Model organisms such as the fruit fly Drosophila melanogaster offer a cost-effective alternative where sample size, environment, and genetic background can be controlled. Cocaine binds to the dopamine transporter of Drosophila as it does in humans, eliciting cocaine-induced locomotor phenotypes. GWAS in Drosophila are facilitated by the Drosophila melanogaster Genetic Reference Panel (DGRP), an expanding collection of wild-derived, inbred, fully sequenced lines that provide a living library of natural variation. It is my goal to use the DGRP and a systems genetics approach to uncover the genetic underpinnings of cocaine-related phenotypes. In Specific Aim 1, I will investigate the impact of genetic background on cocaine preference by screening 600 DGRP lines for cocaine preference using the microplate feeder assay. I will then perform a GWAS, utilizing whole genome sequence data to identify genetic variants associated with cocaine preference. I will perform gene ontology and KEGG orthology pathway enrichment analyses, as well as construct genetic interaction networks to implicate genes and pathways involved in cocaine preference. In Specific Aim 2, I will quantify the behavioral and transcriptomic effects of cocaine exposure in DGRP lines with extreme cocaine preference. For 25 DGRP lines that exhibit cocaine preference and 25 lines near the population average, I will assess effects of cocaine on sensorimotor integration upon repeated exposures. In Specific Aim 3, I will functionally validate candidate genes and variants associated with cocaine preference using RNA interference and out-of-sample testing of DGRP lines, respectively. Completion of these aims will result in a systems genetics model of cocaine preference in Drosophila with translational potential for human cocaine use disorder. This project will train me in advanced systems genetics techniques and analyses and prepare me for a professional career in the study of the genetics of substance use disorders.

项目总结 可卡因使用障碍(CUD)是一个重大的公共卫生和社会经济问题。可卡因是 参与了40%与毒品有关的急诊室就诊，美国可卡因过量使用率为 稳步攀升。然而，导致CUD的基因基础仍然知之甚少。双胞胎研究 关于CUD的遗传学，遗传力估计在42-79%之间。然而，显著的遗传力估计 不要指出哪些基因参与其中，以及它们可能如何在功能上相关。全基因组关联 由于不能控制环境因素，CUD的研究受到限制；单倍型结构 人类基因组作图精度有限；由于招募参与者困难，样本量较小 因为可卡因的使用被定为刑事犯罪。果蝇等果蝇等模式生物提供 一种可控制样本大小、环境和遗传背景的经济高效的替代方案。可卡因 与人类一样与果蝇的多巴胺转运体结合，引发可卡因诱导的运动 表型。果蝇黑腹果蝇遗传参考小组促进果蝇的GWAs (DGRP)，这是一个不断扩大的野生来源、近交系、全测序品系的集合，提供了一个活的 自然变异。我的目标是使用DGRP和系统遗传学的方法来揭示 可卡因相关表型的基础。在特定的目标1中，我将研究基因的影响 用微孔板筛选600个可卡因偏爱DGRP株系的可卡因偏爱背景 饲喂试验。然后，我将利用全基因组序列数据来识别遗传变异，进行GWA 与可卡因偏好有关。我将进行基因本体论和KEGG正交学途径丰富 分析，以及构建遗传相互作用网络，以涉及与可卡因有关的基因和途径 偏好。在具体目标2中，我将量化暴露在可卡因环境中的行为和转录影响 对可卡因有极端偏好的DGRP品系。对于25个表现出可卡因偏好的DGRP品系和25个品系 在接近人群平均水平时，我将评估可卡因对感觉运动整合的影响 曝光。在具体目标3中，我将从功能上验证与可卡因相关的候选基因和变种 偏好分别使用RNA干扰和DGRP品系的样本外测试。完成这些工作 AIMS将导致果蝇对可卡因偏好的系统遗传学模型，该模型具有翻译 人类可卡因使用障碍。这个项目将训练我先进的系统遗传学技术和分析。 并为我在物质使用障碍的遗传学研究方面的职业生涯做好准备。