Systems genetics of premorbid and cocaine use traits in a rat reduced complexity cross

降低复杂性杂交大鼠病前和可卡因使用特征的系统遗传学

基本信息

批准号：
10375811
负责人：
CAMRON D BRYANT
金额：
$ 73.79万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2027-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10375811
关键词：
Behavior Behavioral Brain Brain region Candidate Disease Gene Chromosome Mapping Cocaine Cocaine Dependence Complex DNA Data Set Exons FDA approved Female Future Genes Genetic Genetic Crosses Genotype Heritability Impulsivity Inbred SHR Rats Inbred Strain Inbreeding Intake Lead Maps Messenger RNA Methamphetamine Modeling Molecular Motor Activity Mus Mutation Neurobiology Neuronal Plasticity Nucleus Accumbens Pathway Analysis Pharmaceutical Preparations Phenotype Prefrontal Cortex Proteins Proteomics Public Health Quantitative Trait Loci Rattus Research Personnel Rewards Risk Risk Factors Rodent Saline Signal Transduction Sucrose System Therapeutic Training Transcript Transcriptional Regulation Translations Validation Variant addiction base behavioral phenotyping biomarker panel brain tissue causal variant cocaine self-administration cocaine use design differential expression drug action forward genetics functional genomics genetic approach genetic variant genome sequencing genome wide association study insertion/deletion mutation insight male multiple omics mutation correction novel stimulant use disorder trait transcriptome transcriptome sequencing transcriptomics translation to humans whole genome

项目摘要

PROJECT SUMMARY Stimulant use disorders (e.g., cocaine, methamphetamine) are a major public health concern. Despite a heritability of ~40-50%, genome-wide association studies (GWAS) have identified very few loci, including one hit for cocaine (COC) dependence that maps to FAM53B, a gene also identified via expression quantitative trait locus (QTL) analysis to be associated with COC self-administration in mice. The primary objective is to rapidly identify novel genetic factors in rats that contribute to premorbid risk (compulsivity, impulsivity) and cocaine use traits in a spontaneously hypertensive rat (SHR) reduced complexity cross (RCC). A rodent systems genetics approach triangulates on discovery-based genetic and multi-level functional genomic analysis and can provide a more rapid genetic and neurobiological insight into drug action and neuroplasticity underlying addiction. For several years, the contact PI has been employing mouse reduced complexity crosses (RCCs) between near-isogenic inbred substrains to facilitate gene mapping, validation, and mechanisms. Because rodent substrains are > 99% genetically identical and contain several orders of magnitude fewer variants compared to classical inbred strains, mapping quantitative trait loci (QTLs) in RCCs yields orders of magnitude fewer causal candidate genes to consider. When combined with functional genomics, RCCs can rapidly lead to causal gene and variant identification. Our preliminary studies establish robust, heritable differences in premorbid impulsivity and compulsivity, sucrose reward sensitivity, and multiple COC use traits between SHR/NCrl and SHR/NHsd substrains, including COC-induced locomotor activity, COC IVSA taking, seeking, and intake cycles, demonstrating feasibility for gene mapping in an RCC. In Aims 1 and 2, we will pioneer the use of a rat RCC where we will conduct whole genome sequencing (WGS) and map behavioral QTLs and expression QTLs (eQTLs) from nucleus accumbens (NAC) and prefrontal cortex (PFC) at the whole transcript and exon levels in an F2 cross comprising COC-trained versus yoked saline (SAL)-trained rats. In Aim 3, we will conduct proteomic analysis of PFC and NAC from COC vs. yoked SAL-trained rats to triangulate on high confidence candidate quantitative trait genes (QTGs) and variants (QTVs) as we build functional connections between DNA variants, transcriptional regulation, protein translation, and cell signaling adaptations underlying premorbid and cocaine use traits. These studies pioneer the use of a rat RCC combined with deep behavioral phenotyping to rapidly identify high-confidence candidate novel genetic factors and molecular mechanisms influencing premorbid risk factors and cocaine use traits. Future gene editing of candidate causal gene variants will be modeled on the two near-isogenic SHR backgrounds to demonstrate necessity (mutation correction; “rescue”) and sufficiency (mutation induction). Deliverables include WGS’s of SHR substrains for future RCCs for complex trait analysis as well as adaptive rat transcriptomic and proteomic datasets in key brain regions of the mesocorticolimbic circuitry that can be further mined by investigators and hopefully inform therapeutics.

项目概要尽管兴奋剂使用障碍（例如可卡因、甲基苯丙胺）是一个主要的公共卫生问题。遗传力约为 40-50%，全基因组关联研究 (GWAS) 已识别出极少数位点，包括一个命中位点可卡因 (COC) 依赖映射到 FAM53B，该基因也通过表达数量性状鉴定与 COC 小鼠自我给药相关的基因座 (QTL) 分析的主要目标是快速进行。识别大鼠中导致病前风险（强迫性、冲动性）的新遗传因素，以及自发性高血压大鼠 (SHR) 降低复杂性交叉 (RCC) 中的可卡因使用特征。系统遗传学方法对基于发现的遗传和多级功能基因组分析进行三角测量并且可以提供对药物作用和神经可塑性的更快速的遗传和神经生物学见解多年来，联络 PI 一直采用小鼠简化复杂性杂交 (RCC)。近等基因近交亚系之间的关系，以促进基因定位、验证和机制。啮齿动物亚系的基因相同性 > 99%，且变异数少几个数量级与经典近交系相比，绘制 RCC 中数量性状位点 (QTL) 的产量达到了数量级当与功能基因组学结合时，需要考虑的因果候选基因更少，RCC 可以迅速导致。我们的初步研究证实了病前的强大的、可遗传的差异。 SHR/NCrl 和 SHR/NCrl 之间的冲动性和强迫性、蔗糖奖励敏感性以及多种 COC 使用特征 SHR/NHsd 亚系，包括 COC 诱导的运动活动、COC IVSA 服用、寻找和摄入周期，证明 RCC 基因图谱的可行性在目标 1 和 2 中，我们将率先使用大鼠 RCC。我们将进行全基因组测序 (WGS) 并绘制行为 QTL 和表达 QTL （eQTL）来自伏隔核（NAC）和前额皮质（PFC）的整个转录本和外显子水平由 COC 训练与带轭盐水 (SAL) 训练的大鼠组成的 F2 杂交在目标 3 中，我们将进行蛋白质组学研究。对 COC 与带轭 SAL 训练大鼠的 PFC 和 NAC 进行分析，以对高置信度候选者进行三角测量当我们在DNA变异之间建立功能联系时，性状基因（QTG）和数量变异（QTV），病前和可卡因的转录调控、蛋白质翻译和细胞信号适应这些研究开创了将大鼠 RCC 与深度行为表型分析相结合的方法。确定影响病前风险的高可信度候选新遗传因素和分子机制候选因果基因变异的未来基因编辑将以这两者为模型。近等基因 SHR 背景以证明必要性（突变校正；“救援”）和充分性（突变诱导）包括 SHR 亚品系的全基因组测序（WGS），用于未来 RCC 的复杂性状分析。以及中皮质边缘关键大脑区域的适应性大鼠转录组和蛋白质组数据集研究人员可以进一步挖掘这些电路，并有望为治疗提供信息。