The Role of lncRNA MALAT1 in Controlling Behavior and Neuroinflammation in Alcohol Use Disorder

项目摘要 酒精使用障碍(AUD)在美国是一种流行的精神疾病，给人们带来了巨大的 这给社会造成了巨大损失，每年造成数千人死亡。虽然许多行为变化 与这种疾病相关的特征是，潜在的生理状况，如 神经炎症和转录改变，需要进一步研究。这项提议提出了一部小说 检验协调这些分子和行为变化的潜在机制的假说(S)。 结合以前的文献，我假设长的非编码RNA(LncRNA)MALAT1可能在 对酒精和酒精消费的神经炎性反应。 非编码RNA在几种基本细胞中继续被认为是不可或缺的角色 功能。MALAT1已被证明协调基因表达的转录调控和炎症 过程，多项研究表明MALAT1和高迁移率组框1之间存在联系 (HMGB1)/Toll样受体4(TLR4)神经炎性级联反应。在这项研究计划中，我想确定 如果MALAT1调节HMGB1/TLR4通路的表达以响应乙醇，特别是在星形胶质细胞中， 中枢神经系统细胞类型，调节神经免疫反应，在控制行为中起关键作用。第一 这项建议的目的是利用CRISPR/Cas9在原代培养的星形胶质细胞中下调MALAT1的表达 并评估酒精诱导的神经免疫基因表达和细胞因子产生的变化。这个 第二个目标是创造一种新的有牙线的MALAT1条件性基因敲除小鼠，它将被培育成一系列小鼠 重组酶Cre在星形胶质细胞中的选择性表达。这种交配的后代将拥有 星形胶质细胞特异性MALAT1基因敲除。评估酒精诱导的神经免疫反应和酒精 这些动物的饮酒行为将为深入了解MALAT1和星形胶质细胞在调节 澳元病的基本病理。 该项目是了解lncRNA MALAT1在体内功能贡献的重要一步 以及这种疾病的一些生理表型可能如何永久存在。几乎没有什么有效的 AUD患者目前可用的治疗方法，来自拟议研究的结果 可能指向新的预防策略和治疗目标。此外，该项目将极大地增强 我的研究生培训，因为它将给我一个机会来设计和执行假设驱动的研究 令人振奋的新研究领域和掌握创新技术。归根结底，这个Ruth提供的培训 L.Kirschstein国家研究服务奖(F31)将有助于我作为研究生的科学训练，因为 以及作为一名独立调查员的职业发展。

Project Summary Alcohol use disorder (AUD) is a prevalent psychiatric condition in the United States, imposing a huge cost on society and contributing to thousands of deaths each year. While many of the behavioral changes associated with the disorder have been characterized, the underlying physiological conditions, such as neuroinflammation and transcriptomic alterations, require further study. This proposal presents a novel hypothesis for testing the potential mechanism(s) coordinating these molecular and behavioral changes. Integrating previous literature, I hypothesize that the long non-coding RNA (lncRNA) Malat1 may mediate neuroinflammatory responses to ethanol and ethanol consumption. Non-coding RNAs are continually being recognized as integral players in several basic cellular functions. Malat1 has been shown to coordinate transcriptional regulation of gene expression and inflammatory processes, with multiple studies suggesting a link between Malat1 and the high mobility group box 1 (Hmgb1)/Toll-like receptor 4 (Tlr4) neuroinflammatory cascade. In this research proposal, I want to determine if Malat1 regulates expression of the Hmgb1/Tlr4 pathway in response to ethanol, particularly in astrocytes, a CNS cell-type that regulates neuroimmune responses and plays a pivotal role in controlling behavior. The first aim of this proposal will utilize CRISPR/Cas9 to knockdown Malat1 expression in primary astrocyte cultures and assess ethanol-induced changes in neuroimmune gene expression and the production of cytokines. The second aim will create a novel floxed Malat1 conditional knockout mouse which will be bred to a line of mice expressing the recombinase enzyme Cre selectively in astrocytes. Offspring of this mating will possess astrocyte-specific knockout of Malat1. Assessing the ethanol-induced neuroimmune response and ethanol drinking behavior of these animals will provide insight into the role of both Malat1 and astrocytes in regulating the underlying pathology of AUD. This project is an important step in understanding the functional contribution of the lncRNA Malat1 in AUD and how some of the physiological phenotypes of the disorder may be perpetuated. With few effective treatments currently available for individuals suffering from AUD, results obtained from the proposed studies may point to new prevention strategies and therapeutic targets. Additionally, this project will greatly enhance my graduate training, as it will give me the opportunity to design and perform hypothesis driven research in an exciting new area of research and master innovative techniques. Ultimately, the training provided by this Ruth L. Kirschstein National Research Service Award (F31) will aid in my scientific training as a graduate student, as well as in my professional development as an independent investigator.