Role of noncoding RNA in alcohol action

项目摘要 乙醇诱导的脑转录组变化是脑缺血的发生和持续的基础。 酒精使用障碍（AUD）INIA-N的研究人员和其他人发现， 一组高度限制的非编码RNA（ncRNA）发生了特异性和显著的变化， 包括长ncRNA（lncRNA）。 我们证实这些乙醇响应性lncRNA与AUD 脑转录组。 具体目标1将检验单个lncRNA是关键的假设， 酒精饮料的调节器。 为了验证这一假设，基因工程小鼠与改变 将产生并分析lncRNA的表达。 那些改变了饮酒习惯的变异品系 行为将由多个INIA-N研究者仔细检查机制洞察力。 研究脑中ncRNA功能的障碍是缺乏有效的方法， 非侵入性地将ncRNA和/或ncRNA拮抗剂递送至脑的大部分。这里我们 体内给药外泌体（内源性产生的脂质体样纳米颗粒） 可以被利用来在整个大脑中递送ncRNA拮抗剂或模拟物。这种方法可以 也可用于优先向脑递送药物。 这种方法将针对药物 选择性地作用于所需的作用部位（大脑），同时避免限制的外周毒性 疗效因为这种方法具有靶特异性和 无创性，具有巨大的转化潜力。 具体目标2将测试 假设外泌体可以作为有效的ncRNA/药物递送载体来调节 酒精饮料 基因工程啮齿动物允许调查参与假定的乙醇目标 in the context背景of whole整个animal动物behavior行为responses反应. 因为关于假定的 乙醇目标必须最终解释乙醇诱导的行为表型，整个动物 实验代表了最严格的相关性测试。 为此，Specific Aim 3将创建 INIA-N和INIA-Stress研究者的设计小鼠。基因工程动物将成为 使用最先进的CRISPR/Cas9基因编辑技术生产。 这一合作具体 目的是继续INIA-West基因工程啮齿动物核心项目，该项目在2008年期间获得资助。 项目前期。

Project Summary Ethanol-induced changes in the brain transcriptome underlie the development and persistence of alcohol use disorder (AUD). INIA-N investigators and others have discovered that ethanol induces specific and dramatic alterations in a highly restricted group of noncoding RNAs (ncRNAs) that includes long ncRNAs (lncRNAs). We posit that these ethanol responsive lncRNAs coordinate AUD brain transcriptomes. Specific Aim 1 will test the hypothesis that individual lncRNAs are key regulators of ethanol drinking. To test this hypothesis, genetically engineered mice with altered expression of lncRNAs will be created and analyzed. Those mutant lines with altered drinking behavior will be scrutinized for mechanistic insight by multiple INIA-N investigators. A barrier to the study of ncRNA function in brain is the dearth of efficient methods of noninvasively delivering ncRNAs and/or ncRNA antagonists to large portions of the brain. Here we posit that intransally administered exosomes (endogenously produced, liposome like nanoparticles) can be harnessed to deliver ncRNA anatagonists or mimics throughout the brain. This approach can also be used to deliver drugs preferentially to brain. Such an approach would target drugs selectively to the desired site of action (brain) while avoiding peripheral toxicities that limit therapeutic efficacy. Because this approach has the dual benefits of target specificity and noninvasiveness, it has tremendous translational potential. Specific Aim 2 will test the hypothesis that exosomes can be harnessed as effective ncRNA/drug delivery vehicles to modulate ethanol drinking. Genetically engineered rodents permit investigation of the involvement of putative ethanol targets in the context of whole animal behavioral responses. Because hypotheses concerning putative ethanol targets must ultimately explain ethanol-induced behavioral phenotypes, whole-animal experiments represent the most rigorous test of relevance. To this end, Specific Aim 3 will create designer mice for both INIA-N and INIA-Stress investigators. Genetically engineered animals will be produced using state of the art CRISPR/Cas9 gene editing technology. This collaborative Specific Aim is the continuation of the INIA-West Genetically Engineered Rodents Core that was funded during the previous project period.