3/11 Epigenetic Regulation of Neuroimmune Pathways

3/11 神经免疫途径的表观遗传调控

• 批准号: 10410092
• 负责人: Sean P Farris
• 金额: $ 60.15万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-05 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10410092
• 关键词: Affect; Alcohol consumption; Alcohol dependence; Alcoholism; Animal Behavior; Animals; Biological Process; Candidate Disease Gene; Cell Nucleus; Cell physiology; Cells; Chromatin; Chronic; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collaborations; Collection; Consumption; Development; Disease; Distant; Drug Addiction; Enhancers; Epigenetic Process; Ethanol; Gene Expression; Gene Expression Regulation; Genes; Genetic Engineering; Genome; Genomic Segment; Genomic approach; Genomics; Goals; Hi-C; High-Throughput Nucleotide Sequencing; High-Throughput RNA Sequencing; Human; Immune system; Immunity; In Vitro; Individual; Knock-out; Knockout Mice; LoxP-flanked allele; Maps; Mediating; Microglia; Molecular; Molecular Target; Mus; Mutagenesis; Mutant Strains Mice; Neuraxis; Neuroimmune; Neuroimmune system; Neurosciences; Nucleotides; Organism; Pathway interactions; Peripheral; Proteins; RNA; Regulatory Element; Research; Research Personnel; Research Project Grants; Role; Signal Pathway; Signal Transduction; Space Perception; Structure; System; Systems Development; Tamoxifen; Techniques; Technology; Testing; Training; Transcript; Translating; Untranslated RNA; Validation; alcohol behavior; alcohol exposure; alcohol response; alcohol testing; alcohol use disorder; behavior test; behavioral phenotyping; behavioral response; brain cell; chromosome conformation capture; epigenetic regulation; experimental study; genetic approach; genome editing; genome-wide; genomic locus; in vitro Model; in vivo; mouse Cre recombinase; mutant; negative affect; neuroinflammation; neuropsychiatric disorder; novel; preference; promoter; response; transcriptome

PROJECT SUMMARY Alcohol use disorder (AUD) is a debilitating neuropsychiatric disorder, negatively affecting the lives of millions of individuals worldwide. Chronic ethanol exposure is known to alter multiple molecular pathways throughout the central nervous system, including aberrant neuroimmune signaling. As part of the Integrative Neuroscience Initiative on Alcoholism (INIA) Neuroimmune consortium, the proposed collaborative research project will focus on the contribution of long non-coding RNAs (lncRNAs) to molecular adaptations in the neuroimmune system and development of AUD. The non-coding transcriptome significantly outnumbers the protein-coding transcripts, but the biological function of most non-coding RNAs has yet to be determined. Several studies have suggested that lncRNAs are critical epigenetic regulators of genomic structure and long-term gene expression. We hypothesis that lncRNAs are epigenetic modulators of gene expression in response to ethanol that actively coordinate persistent alterations in cellular function and animal behavior. We have proposed three specific aims to experimentally test this hypothesis and help accomplish the overall goals of the INIA consortium. Aim 1 will use Perturb-Seq to functionally test the involved of lncRNAs in neuroimmune gene expressing using an established in vitro model of ethanol exposure. Combining use of the clustered regularly interspaced short palindromic repeats (CRISPR) genome editing approach with high-throughput RNA- Sequencing this aim will identify neuroimmune pathways regulated by specific lncRNAs. Aim 2 will use chromosome conformation capture sequencing to demonstrate physical changes in the spatial orientation of chromatin (e.g., promoter-enhancer interactions) due to excessive ethanol exposure. This specific aim will create a new genome-wide chromatin interaction map that will identify regulatory elements involved in the ethanol-responsive neuroimmune pathways. Aim 3 will use CRISPR-mediated genome editing in vivo strategies to determine the role of individual lncRNAs in regulating neuroimmune activation and ethanol-related behavioral phenotypes. This specific aim will create several novel genetically engineered lines of mice, for multiple collaborative projects, to test ethanol-related behaviors and molecular mechanisms associated with excessive ethanol exposure. Completion of the proposed research will broaden our understanding for epigenetic regulation of the neuroimmune system in AUD and determine molecular adaptations underlying excessive ethanol exposure.

项目摘要 酒精使用障碍（AUD）是一种令人衰弱的神经精神障碍，对数百万的生活产生了负面影响 全球个人。已知慢性乙醇暴露会改变整个过程中的多个分子途径 中枢神经系统，包括异常的神经免疫信号传导。作为综合神经科学的一部分 拟议的合作研究项目将针对酒精中毒（INIA）神经免疫财团（INIA）倡议 关于长的非编码RNA（LNCRNA）对神经免疫系统中分子适应的贡献 和AUD的开发。非编码转录组显着超过蛋白质编码 转录本，但大多数非编码RNA的生物学功能尚未确定。几项研究 已经表明lncRNA是基因组结构和长期基因的关键表观遗传调节剂 表达。我们假设LNCRNA是对乙醇的反应的基因表达的表观遗传调节剂 该积极协调细胞功能和动物行为的持续改变。我们提出了三个 具体的目的是实验检验这一假设并有助于实现INIA的总体目标 财团。 AIM 1将使用intturb-seq在功能上测试lncRNA在神经免疫基因中的涉及 使用已建立的体外乙醇暴露模型表达。定期组合聚类的使用 带有高通量RNA- 测序该目标将确定由特定LNCRNA调节的神经免疫途径。 AIM 2将使用 染色体构象捕获测序，以证明在空间方向的物理变化 由于乙醇过多而导致的染色质（例如启动子增强剂相互作用）。这个特定的目标 创建一个新的全基因组染色质相互作用图，该图将确定涉及的调节元素 乙醇反应性神经免疫性途径。 AIM 3将在体内使用CRISPR介导的基因组编辑 确定单个LNCRNA在调节神经免疫激活和乙醇相关的策略 行为表型。这个具体目标将创建几种新颖的基因工程线的小鼠，用于 多个协作项目，以测试与乙醇相关的行为和分子机制 乙醇暴露过多。拟议研究的完成将扩大我们对 AUD中神经免疫系统的表观遗传调节并确定分子适应的基础。 乙醇暴露过多。