lncRNA Gas5 dysregulation alters ethanol drinking behavior and ethanol-related phenotypes

lncRNA Gas5失调改变乙醇饮用行为和乙醇相关表型

• 批准号: 10464571
• 负责人: Sonja Lorean Plasil
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-03 至 2023-03-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10464571
• 关键词: Abnormal coordination; Acute; Addictive Behavior; Adult; Alcohol Phenotype; Alcohol consumption; Alcohol dependence; Alcoholism; Alcohols; American; Amygdaloid structure; Animals; Attention; Behavior; Bilateral; Brain; Brain Diseases; Brain region; CRISPR/Cas technology; Cause of Death; Chronic; Clinical; Cocaine; Complex; Consumption; Dependovirus; Development; Disease; Disease Progression; Drug Addiction; Ethanol; Financial compensation; Foundations; Gene Expression; Genetic Transcription; Genome; Glucocorticoid Receptor; Glucocorticoids; Goals; Growth; Guide RNA; Human; Immune; Immune signaling; In Situ Hybridization; Individual; Intake; Knock-out; Laboratories; Learning; Link; Manuscripts; Medial; Mediating; Mental disorders; MicroRNAs; Molecular; Molecular Mechanisms of Action; Motivation; Motor; Mus; Mutagenesis; Mutate; Mutation; Neurobiology; Neuroimmune; Neurons; Neurosciences; Nucleus Accumbens; Operative Surgical Procedures; Outcome; Pathogenesis; Pathway interactions; Patients; Persons; Phenotype; Porifera; Prefrontal Cortex; Process; Psychiatric Diagnosis; RNA; Regulation; Relapse; Research; Research Personnel; Rewards; Role; Sequence Homology; Substance abuse problem; System; Technical Expertise; Techniques; Testing; Training; United States; Untranslated RNA; Withdrawal; Work; Writing; addiction; alcohol behavior; alcohol exposure; alcohol misuse; alcohol relapse; alcohol response; alcohol testing; alcohol use disorder; anxiety-like behavior; biological adaptation to stress; collaborative environment; craving; design; drinking; drinking behavior; emotional distress; executive function; experimental study; human disease; in vivo; insight; interest; neuroinflammation; neuronal growth; new therapeutic target; overexpression; pre-clinical; preference; promoter; response; targeted treatment; training opportunity; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; vapor

Project Summary Alcohol use disorder (AUD) is a chronic, debilitating, and relapsing brain disease. An overwhelming number of people die yearly due to alcohol-related causes; however, the mechanism(s) of action are challenging to decipher and remain largely elusive. Long noncoding RNAs (lncRNAs) are key regulators of the genome; persistent transcriptional changes induced by chronic ethanol exposure is a hypothesized mechanism for AUD development, withdrawal, and relapse. Our laboratory and others have shown that ethanol-responsive lncRNAs have the ability to directly regulate ethanol- related behavior when mutated in vivo, but only a handful have been characterized to date. In this proposal, I will learn and apply state-of-the-art CRISPR/Cas9 techniques to functionally investigate ethanol-responsive lncRNA Gas5 for its regulation of ethanol drinking and ethanol-related behaviors. As ethanol alters gene expression and molecular pathways that regulate neuroinflammation, I want to target an ethanol-responsive and highly-interconnected competing endogenous RNA (ceRNA) lncRNA that may coordinate large endogenous immune networks related to AUD. I will focus on in vivo CRISPR/Cas9 modulation of a specific ‘hub’ lncRNA of interest, growth arrest-specific 5 (Gas5). Gas5 acts as a ceRNA to regulate immune signaling, is significantly and persistently downregulated following chronic intermittent ethanol vapor exposure and has been shown to reduce cocaine-intake, linking Gas5 to substance abuse. It is therefore of substantial interest to test the hypothesis that Gas5 is a key determinant of ethanol action. Gas5 will be knocked out in a temporal, cellular, and brain region-specific manner to investigate its role on ethanol drinking. Upon completion of this project, I will have advanced our understanding for the molecular impact of ethanol- responsive lncRNA Gas5 and how it relates to addictive behavior. This project will provide substantial training opportunities, a strong research foundation applicable to a variety of scientific fields, and a collaborative environment with other researchers in the field including the Integrative Neuroscience Initiative on Alcoholism – Neuroimmune consortium. A wealth of technical skills ranging from bench work to complex surgical techniques, the understanding and development of project design, and several scientific writing manuscript prospects are available within each aim that will be valuable for my continued scientific training.

项目摘要 酒精使用障碍（AUD）是一种慢性、衰弱和复发性脑部疾病。绝大多数 每年都有人死于与酒精有关的原因;然而，其作用机制难以破译 并且在很大程度上仍然难以捉摸 长链非编码RNA（lncRNA）是基因组的关键调控因子; 慢性乙醇暴露是AUD发展、戒断和复发的假设机制。我们 实验室和其他机构已经表明，乙醇响应性lncRNA具有直接调节乙醇的能力- 相关的行为时，在体内突变，但只有少数已被确定的日期。在这份提案中，我 将学习和应用最先进的CRISPR/Cas9技术来功能性地研究乙醇响应性 lncRNA Gas 5对饮酒和乙醇相关行为的调节。 由于乙醇改变了调节神经炎症的基因表达和分子途径，我想靶向一个神经元， 乙醇响应性和高度互连的竞争性内源性RNA（ceRNA）lncRNA， 协调与AUD相关的大型内源性免疫网络。 我将专注于体内CRISPR/Cas9对感兴趣的特定“枢纽”lncRNA的调节，生长停滞特异性5 （Gas5）。Gas 5作为一种ceRNA调节免疫信号传导，被显著且持续下调 在慢性间歇性乙醇蒸汽暴露后，已被证明可以减少可卡因摄入量， 5、滥用药物。因此，检验Gas 5是一个关键的假设是非常有意义的。 乙醇作用的决定因素。Gas 5将以时间、细胞和大脑区域特异性的方式被敲除 来研究它对酒精饮料的影响 完成这个项目后，我将进一步了解乙醇的分子影响- 反应性lncRNA Gas 5以及它与成瘾行为的关系。该项目将提供大量培训 机会，适用于各种科学领域的强大研究基础，以及合作 环境与其他研究人员在该领域，包括综合神经科学倡议酒精中毒- 神经免疫联盟。丰富的技术技能，从板凳工作到复杂的手术技术， 项目设计的理解和发展，以及几篇科学论文的写作展望， 在每一个目标，这将是有价值的，我继续科学训练。