Role of glial expression in nicotine behaviors for genes identified through human GWAS

通过人类 GWAS 鉴定的基因神经胶质表达在尼古丁行为中的作用

基本信息

批准号：
10542587
负责人：
MARISSA A EHRINGER
金额：
$ 17.35万
依托单位：
UNIVERSITY OF COLORADO
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：
2022-09-15 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10542587
关键词：
Abstinence Acute Address Affect Animal Model Animal Testing Animals Antisense Oligonucleotides Anxiety Astrocytes Behavior Biochemical Biochemical Markers Biological Biological Assay Body Temperature Brain Brain region Candidate Disease Gene Cell Culture Techniques Chronic Data Development Event Excision Exhibits Family Future Gene Expression Profile Generations Genes Genetic Genetic Transcription Genotype Health Heart Rate Histologic Human Human Genome In Vitro Individual Intake Knockout Mice Knowledge Laboratories Lead Learning Measures Modeling Morphology Motivation Motor Activity Mouse Strains Mus Neurobiology Neurons Nicotine Nicotine Dependence Nicotine Withdrawal Nicotinic Receptors Pathway Analysis Pathway interactions Phase Phenotype Play Prevention strategy Public Health Publishing RNA Reagent Research Rewards RiboTag Role Sex Differences Signal Transduction Site Sleep Small Interfering RNA Smoking Smoking Behavior Societies Specificity Substance abuse problem Technology Testing Tobacco Tobacco use Translating United States Variant Vendor Withdrawal Withdrawal Symptom Woman base behavioral response brain cell conditional knockout conditioned place preference differential expression drug of abuse druggable target effective therapy electronic cigarette use gene network genome wide association study genome-wide improved in vivo in vivo Model innovation insight interest knock-down mouse model nicotine exposure nicotine treatment nicotine use novel relating to nervous system response smoking cessation smoking relapse transcriptome transcriptomics

项目摘要

PROJECT SUMMARY Tobacco use rates, including the increasing use of electronic cigarettes, remain at ~20% in the United States, leading to long-term health and financial consequences for individuals, families, and society. Many individuals who desire to quit smoking cannot stop, and the underlying neurobiological effects of smoking on the brain remain poorly understood. Recent large-scale genome-wide association studies (GWAS) have provided convincing evidence for the association of over 2500 loci with smoking behaviors. Understanding the biological mechanisms through which these genes impact smoking behavior may provide novel insights into the underlying biological mechanisms responsible for the use of tobacco products. It is well known that nicotine is the main psychoactive component in tobacco and is responsible for many of the effects of smoking, from initial reward to many aspects of withdrawal. Its mechanism of action generally is attributed to its action on nicotinic acetylcholine receptors (nAChRs) on neurons. However, astrocytes also express nAChRs and may play a role in nicotine dependence. Accumulating evidence indicates that astrocytes actively participate in behavioral responses to nicotine and nicotine abstinence in both animal models and humans. Therefore, some genes associated with nicotine behaviors in humans may impact these behaviors by altering astrocyte function. Because little is known about the role of astrocytes in nicotine behaviors, this project aims to use an astrocyte in vitro functional assay in the R21 phase to functionally assess and prioritize nicotine GWAS genes that may contribute to nicotine behaviors through their actions in astrocytes. The genes of interest will be validated in vivo following nicotine exposure as an initial GO NO-GO for that gene. Conditional knock-out (cKO) mouse models will then be made for the top two genes identified through the screen, and the GO NO-GO decision will be based on the selectivity and efficiency of removal from astrocytes. The R33 phase will utilize the cKO mice to confirm functional astrocyte effects of the selected genes and assess the specific nicotine phenotypes impacted by targeting these genes selectively in astrocytes. Finally, appropriate brain regions from animals tested will be used for transcriptome studies to identify novel genes and pathways in astrocytes differentially affected by genotype or nicotine exposure. Our approach is conceptually and technically innovative because we will be the first to test the idea that astrocyte culture can be used to functionally assess the role of GWAS- identified genes in nicotine-related neurobiology. We will utilize novel genetic reagents to target GOIs in astrocytes in the brain specifically. This study is also the first to use transcriptomics to address questions of GOI function in nicotinic neural responses, specifically in astrocytes. The proposed research is significant because it will provide insight into astrocyte and GWAS identified gene mechanisms related to nicotine use, which has important implications for improved understanding of nicotine neurobiology and developing more effective therapies to help reduce nicotine use and withdrawal symptoms.

项目概要在美国，烟草使用率（包括电子烟使用量的增加）保持在 20% 左右，给个人、家庭和社会带来长期的健康和经济后果。很多个人想要戒烟的人无法停止，以及吸烟对大脑的潜在神经生物学影响仍然知之甚少。最近的大规模全基因组关联研究（GWAS）提供了超过 2500 个基因座与吸烟行为相关的令人信服的证据。了解生物学这些基因影响吸烟行为的机制可能为研究吸烟行为提供新的见解导致烟草制品使用的潜在生物机制。众所周知，尼古丁是烟草中主要的精神活性成分，是造成吸烟从最初开始的许多影响的原因对退出的奖励是多方面的。其作用机制一般归因于其对烟碱的作用神经元上的乙酰胆碱受体（nAChR）。然而，星形胶质细胞也表达 nAChR，并可能发挥作用尼古丁依赖。越来越多的证据表明星形胶质细胞积极参与行为动物模型和人类对尼古丁和尼古丁戒断的反应。因此，有些基因与人类尼古丁行为相关的物质可能通过改变星形胶质细胞功能来影响这些行为。由于人们对星形胶质细胞在尼古丁行为中的作用知之甚少，因此该项目旨在使用星形胶质细胞 R21 阶段的体外功能测定，对尼古丁 GWAS 基因进行功能评估和优先排序，这些基因可能通过在星形胶质细胞中的作用促进尼古丁行为。感兴趣的基因将在体内尼古丁暴露作为该基因的初始 GO NO-GO。条件性基因敲除（cKO）小鼠然后将为通过筛选确定的前两个基因建立模型，并做出 GO/NO-GO 决策基于从星形胶质细胞去除的选择性和效率。 R33阶段将利用cKO小鼠确认所选基因的功能性星形胶质细胞效应并评估特定的尼古丁表型通过选择性地靶向星形胶质细胞中的这些基因来影响。最后，动物的适当大脑区域测试将用于转录组研究，以差异性地识别星形胶质细胞中的新基因和途径受基因型或尼古丁暴露的影响。我们的方法在概念和技术上都是创新的，因为我们将成为第一个测试星形胶质细胞培养可用于功能性评估 GWAS 作用的想法的人鉴定了尼古丁相关神经生物学中的基因。我们将利用新型遗传试剂来靶向 GOI 特别是大脑中的星形胶质细胞。这项研究也是第一个使用转录组学来解决以下问题的研究 GOI 在烟碱神经反应中发挥作用，特别是在星形胶质细胞中。拟议的研究意义重大因为它将提供对星形胶质细胞和 GWAS 确定的与尼古丁使用相关的基因机制的深入了解，这对于提高对尼古丁神经生物学的理解并开发更多尼古丁神经生物学具有重要意义有助于减少尼古丁使用和戒断症状的有效疗法。