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; 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.

项目摘要 可卡因使用障碍（CUD）代表了一个重大的公共卫生和社会经济问题。可卡因是 涉及40％与毒品有关的急诊室就诊，美国可卡因过量的比率是 稳步攀登。然而，易感cud的遗传基础知识仍然很少。双研究 关于CUD的遗传学，可遗传性估计在42-79％之间。但是，大量的遗传力估计值 请勿表明涉及哪些基因，以及它们如何在功能上相关。全基因组关联 CUD的研究（GWAS）受到无法控制环境因素的限制。单倍型结构 人类基因组限制映射精度；由于难以招募参与者而引起的小样本量 由于可卡因的使用定罪。诸如果蝇果蝇（Drosophila Melanogaster）等模型生物提供 可以控制样本量，环境和遗传背景的一种成本效益的替代方案。可卡因 与人类一样，与果蝇的多巴胺转运蛋白结合，引起可卡因诱导的运动 表型。果蝇中的GWAS由果蝇Melanogaster遗传参考面板促进 （DGRP），野生衍生的，近交，完整的线条的收藏，提供了一个活着的库 自然变化。我的目标是使用DGRP和系统遗传学方法来发现遗传 可卡因相关表型的基础。在特定目标1中，我将研究遗传的影响 通过使用微板筛选可卡因偏好的600 DGRP线的可卡因偏好背景 馈线分析。然后，我将使用整个基因组序列数据进行GWAS，以识别遗传变异 与可卡因偏好相关。我将执行基因本体论和KEGG矫正途径富集 分析以及构造遗传相互作用网络，暗示与可卡因有关的基因和途径 偏爱。在特定目标2中，我将量化可卡因暴露在 具有极端可卡因偏好的DGRP线。对于表现出可卡因偏好和25行的25种DGRP线 接近人口平均水平，我将评估可卡因对重复的感觉运动积分的影响 暴露。在特定目标3中，我将在功能上验证与可卡因相关的候选基因和变体 使用RNA干扰和DGRP系的样本外测试的偏好。这些完成 目标将导致果蝇中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。该项目将在高级系统遗传技术和分析中训练我 并为我的职业生涯做好准备，以研究物质使用障碍的遗传学。