Epigenomic regulation of oxidative stress-producing innate immunity in neuroinflammation

神经炎症中氧化应激产生的先天免疫的表观基因组调控

• 批准号: 10429847
• 负责人: Andrew S Mendiola
• 金额: $ 12.09万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-15 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10429847
• 关键词: Advisory Committees; Binding; Biological Assay; CRISPR interference; California; Candidate Disease Gene; Cells; ChIP-seq; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Coculture Techniques; Communities; Complex; Coupled; Data; Data Set; Development; Disease; Disease Progression; Elements; Enhancers; Environment; Epigenetic Process; Experimental Autoimmune Encephalomyelitis; Experimental Designs; Faculty; Failure; Fluorescent in Situ Hybridization; Foundations; Future; Gene Expression Profile; Gene Targeting; Genes; Genetic Transcription; Genome engineering; Genomics; Goals; Histones; Immune; Immune System Diseases; Immune response; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Innate Immune Response; Innate Immune System; Institutes; Intervention; Investigation; Laboratories; Laboratory Research; Leadership; Link; Macrophage Activation; Maps; Mediating; Mentors; Methods; Microglia; Modeling; Molecular; Multiple Sclerosis; Mus; NADPH Oxidase; Natural Immunity; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurons; Nucleic Acid Regulatory Sequences; Outcomes Research; Oxidative Regulation; Oxidative Stress; Pathogenesis; Pathogenicity; Pathologic; Peripheral; Pharmaceutical Preparations; Phase; Phenotype; Prevention; Process; Production; RNA; Reactive Oxygen Species; Regulator Genes; Regulatory Element; Relapse; Reporter; Research; Resources; Role; San Francisco; Signal Pathway; Spinal Cord; Testing; Therapeutic; Training; Translating; Transposase; Universities; base; biological adaptation to stress; brain tissue; career; career development; cell type; chromatin immunoprecipitation; epigenome; epigenomics; gene repression; genetic signature; genomic locus; histone modification; immune function; in vivo; innovation; loss of function; macrophage; mind control; mouse model; multiple sclerosis treatment; nervous system disorder; neuroinflammation; neurotoxic; neurovascular; novel; novel therapeutic intervention; novel therapeutics; oxidative damage; prevent; programs; promoter; single molecule; therapeutic candidate; therapeutic target; transcription factor

PROJECT SUMMARY / ABSTRACT Oxidative stress is a central part of innate immune-induced neurodegeneration in neurological disorders including multiple sclerosis (MS). However, the molecular mechanisms regulating oxidative stress gene circuits to promote neurotoxic immune responses remain poorly characterized. Emerging evidence supports a role for the epigenome in tightly regulating immune cell gene activity in MS. Yet, the epigenomic landscape and function in prooxidant, neurotoxic central nervous system (CNS) innate immune cells in MS remains unknown. Thus, discovery of drugs capable of selectively suppressing immune-driven neurodegeneration has been hindered by lack of molecular understanding of neurotoxic functions of CNS innate immune cells. The ultimate goal of this project is to define the regulatory landscape of prooxidant immune cells and identify mechanisms that translate epigenetic aberrations into innate-immune driven neurodegeneration for devising novel therapeutic interventions for MS. Our preliminary data discovered a molecular convergence of neurotoxic microglia and peripheral macrophages to a core oxidative stress gene signature in MS model. By applying an innovative experimental design with cutting-edge methods, this proposal aims to define the epigenetic and transcriptional components of oxidative stress-producing innate immune cells in a mouse model of neuroinflammation for MS through unbiased profiling of the open chromatin landscapes (Aim 1) and histone modifications (Aim 2). These molecular characterizations will identify key MS-related regulatory elements that will be functionally validated in vitro and in vivo with CRISPR interference assays (Aim 3). This project will provide a foundational epigenomic outlook on the molecular circuits governing prooxidant, neurotoxic immune responses in neuroinflammatory disease, and the research outcomes may reveal candidates for the development of new treatments for innate immune- mediated oxidative injury in MS and related conditions. The comprehensive training plan will enable the PI to achieve his career goal of launching a successful independent research laboratory dedicated to studying epigenomic mechanisms contributing to immune dysfunction in MS for targeted treatments. The MOSAIC UE5 mentoring, leadership, and diversity training will facilitate his transition to independence and enable the PI to enhance diversity in the biomedical workforce in the R00 phase and beyond. As a mentee in Dr. Katerina Akassoglou’s laboratory, a leader in neurovascular and immune mechanisms of MS pathogenesis, at the esteemed academic environment of Gladstone Institutes and University of California, San Francisco, the PI will obtain new training in functional epigenomics and CRISPR genome engineering during the K99 phase. The PI’s training and career development will be bolstered through an advisory committee of faculty with related expertise; and the PI’s participation in didactic activities such as coursework in epigenomics and seminars, will collectively allow the PI to complete this project and integrate these approaches for making meritorious contributions to the fields of MS and epigenomics in future independent research.

项目概要/摘要 氧化应激是神经系统疾病中先天免疫诱导的神经变性的核心部分 包括多发性硬化症（MS）。然而，调节氧化应激基因回路的分子机制 促进神经毒性免疫反应的机制仍知之甚少。新出现的证据支持 表观基因组在多发性硬化症中严格调节免疫细胞基因活性。然而，表观基因组景观和功能 中枢神经系统（CNS）先天免疫细胞中的促氧化剂、神经毒性物质在多发性硬化症中的作用尚不清楚。因此， 能够选择性抑制免疫驱动的神经变性的药物的发现受到了阻碍 缺乏对中枢神经系统先天免疫细胞的神经毒性功能的分子理解。此次活动的最终目的 该项目的目的是定义促氧化免疫细胞的调控格局，并确定转化的机制 表观遗传畸变导致先天免疫驱动的神经变性，以设计新的治疗干预措施 对于女士。我们的初步数据发现神经毒性小胶质细胞和外周细胞的分子融合 MS 模型中巨噬细胞对核心氧化应激基因特征的影响。通过应用创新实验 采用尖端方法设计，该提案旨在定义表观遗传和转录成分 通过无偏倚在多发性硬化症神经炎症小鼠模型中产生氧化应激的先天免疫细胞 开放染色质景观（目标 1）和组蛋白修饰（目标 2）的分析。这些分子 表征将确定与 MS 相关的关键调控元件，这些元件将在体外进行功能验证 体内 CRISPR 干扰测定（目标 3）。该项目将提供基本的表观基因组前景 神经炎症疾病中控制促氧化剂、神经毒性免疫反应的分子回路，以及 研究结果可能揭示开发先天免疫新疗法的候选者 介导多发性硬化症及相关病症中的氧化损伤。全面的培训计划将使 PI 能够 实现他的职业目标，即创办一个成功的独立研究实验室，致力于研究 导致多发性硬化症免疫功能障碍的表观基因组机制，用于靶向治疗。 UE5马赛克 指导、领导力和多元化培训将有助于他向独立过渡，并使 PI 能够 增强 R00 阶段及以后生物医学劳动力的多样性。作为卡特琳娜博士的学员 Akassoglou 的实验室是多发性硬化症发病机制的神经血管和免疫机制领域的领导者， 格拉德斯通研究所和加州大学旧金山分校享有盛誉的学术环境，PI 将 在 K99 阶段获得功能表观基因组学和 CRISPR 基因组工程方面的新培训。 PI 的 将通过一个由具有相关专业知识的教师组成的咨询委员会来支持培训和职业发展； PI 参与教学活动，例如表观基因组学课程和研讨会，将共同 允许 PI 完成该项目并整合这些方法，为 未来独立研究的 MS 和表观基因组学领域。