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.

项目概要 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 40% 的与毒品相关的急诊室就诊涉及可卡因，美国的可卡因过量服用率是 稳步攀登。然而，人们对导致 CUD 的遗传基础仍然知之甚少。双胞胎研究 对 CUD 遗传学的研究提供了 42-79% 之间的遗传力估计。然而，显着的遗传力估计 没有表明涉及哪些基因，以及它们如何在功能上相关。全基因组关联 CUD 的研究（GWAS）因无法控制环境因素而受到限制；单倍型结构 人类基因组限制绘图精度；由于招募参与者困难而导致样本量较小 因为可卡因的使用被定为刑事犯罪。果蝇等模式生物提供 一种具有成本效益的替代方案，可以控制样本量、环境和遗传背景。可卡因 与人类一样与果蝇的多巴胺转运蛋白结合，引发可卡因诱导的运动 表型。果蝇遗传参考面板促进了果蝇的 GWAS （DGRP），一个不断扩大的野生来源、近交系、完全测序品系的集合，提供了一个活的图书馆 自然变异。我的目标是使用 DGRP 和系统遗传学方法来揭示遗传 可卡因相关表型的基础。在具体目标 1 中，我将研究遗传因素的影响 通过使用微孔板筛选 600 个 DGRP 系的可卡因偏好来了解可卡因偏好的背景 饲养实验。然后我将进行 GWAS，利用全基因组序列数据来识别遗传变异 与可卡因偏好有关。我将进行基因本体和KEGG直系同源通路富集 分析并构建遗传相互作用网络以暗示可卡因涉及的基因和途径 偏爱。在具体目标 2 中，我将量化可卡因暴露对行为和转录组的影响 DGRP 系列对可卡因极度偏爱。对于表现出可卡因偏好的 25 个 DGRP 品系和 25 个品系 接近人口平均水平，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，我将在功能上验证与可卡因相关的候选基因和变异体 分别优选使用 RNA 干扰和 DGRP 系的样本外测试。完成这些 目标将产生果蝇可卡因偏好的系统遗传学模型，具有转化潜力 人类可卡因使用障碍。该项目将培训我先进的系统遗传学技术和分析 并为我从事物质使用障碍遗传学研究的职业生涯做好准备。