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

• 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
• 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

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.

项目摘要 在美国 导致对个人，家庭和社会的长期健康和财务后果。许多人 想要戒烟的人无法停止，吸烟对大脑的潜在神经生物学影响 保持不当理解。最近的大型全基因组关联研究（GWAS）提供了 令人信服的证据证明了超过2500个基因座与吸烟行为的关系。了解生物学 这些基因影响吸烟行为的机制可能会提供新的见解 负责使用烟草产品的基本生物学机制。众所周知，尼古丁是 烟草中的主要精神活性成分，从初始吸烟造成了许多吸烟的影响 奖励撤离的许多方面。它的作用机制通常归因于其对烟碱的作用 神经元上的乙酰胆碱受体（NACHRS）。但是，星形胶质细胞也表达NACHRS，并可能起作用 在尼古丁依赖性中。积累证据表明星形胶质细胞积极参与行为 在动物模型和人类中，对尼古丁和尼古丁的禁欲的反应。因此，一些基因 与人类的尼古丁行为相关，可能会通过改变星形胶质细胞功能来影响这些行为。 因为对星形胶质细胞在尼古丁行为中的作用知之甚少，所以该项目旨在使用星形胶质细胞 在R21阶段的体外功能测定，以评估并优先考虑可能 通过星形胶质细胞的作用来促进尼古丁行为。感兴趣的基因将在 尼古丁暴露后的体内作为该基因的初始go。有条件的敲除（CKO）鼠标 然后，将对通过屏幕标识的前两个基因制作模型，而无需决定将 基于从星形胶质细胞中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 在星形胶质细胞中选择性地靶向这些基因的影响。最后，来自动物的适当大脑区域 测试将用于转录组研究，以鉴定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 我们将是第一个测试星形胶质细胞培养的想法的人，可以在功能上评估GWAS-的作用。 鉴定出与尼古丁相关的神经生物学中的基因。我们将利用新颖的遗传试剂靶向Gois 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的问题 烟碱神经反应中的GOI功能，特别是星形胶质细胞。拟议的研究很重要 因为它将提供有关星形胶质细胞和GWAS鉴定的与尼古丁使用相关的基因机制的见解，所以 这对于改善对尼古丁神经生物学的理解并发展更多具有重要意义 有效减少尼古丁使用和戒断症状的有效疗法。