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.

项目概要 酒精使用障碍 (AUD) 是一种使人衰弱的神经精神疾病，对数百万人的生活产生负面影响 全世界的个人。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 酗酒倡议（INIA）神经免疫联盟，拟议的合作研究项目将重点关注 长非编码 RNA (lncRNA) 对神经免疫系统分子适应的贡献 和澳元的发展。非编码转录组数量显着超过蛋白质编码转录组 转录本，但大多数非编码 RNA 的生物学功能尚未确定。多项研究 表明lncRNA是基因组结构和长期基因的关键表观遗传调节因子 表达。我们假设 lncRNA 是响应乙醇的基因表达的表观遗传调节剂 积极协调细胞功能和动物行为的持续变化。我们提出了三个 具体目标是通过实验检验这一假设并帮助实现 INIA 的总体目标 财团。目标1将使用Perturb-Seq功能测试lncRNA在神经免疫基因中的参与 使用已建立的乙醇暴露体外模型进行表达。定期结合使用集群 间隔短回文重复序列（CRISPR）基因组编辑方法与高通量RNA- 对这一目标进行测序将确定受特定 lncRNA 调节的神经免疫通路。目标2将使用 染色体构象捕获测序来证明空间方向的物理变化 由于过量乙醇暴露而导致染色质（例如启动子-增强子相互作用）。这一具体目标将 创建一个新的全基因组染色质相互作用图谱，该图谱将识别参与染色质相互作用的调控元件 乙醇反应性神经免疫途径。 Aim 3 将使用 CRISPR 介导的体内基因组编辑 确定个体lncRNA在调节神经免疫激活和乙醇相关作用中的作用的策略 行为表型。这一具体目标将创造几种新型基因工程小鼠品系， 多个合作项目，测试与乙醇相关的行为和分子机制 过量的乙醇暴露。完成拟议的研究将拓宽我们对 AUD 中神经免疫系统的表观遗传调控并确定潜在的分子适应 过量的乙醇暴露。