Intracellular nucleic acid detection in autoimmunity

自身免疫中的细胞内核酸检测

• 批准号: 10402241
• 负责人: Daniel B Stetson
• 金额: $ 61.02万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-15 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10402241
• 关键词: ADAR1; Acute; Address; Adenosine; Alleles; Autoimmune; Autoimmunity; Biology; CRISPR screen; Cells; Disease; Double-Stranded RNA; Enzymes; Epitopes; Genes; Host Defense; Human; Impairment; Interferon Type I; Interferon-beta; Interferons; Knock-in; Knock-in Mouse; Ligands; Mus; Mutation; Natural Immunity; Nucleic Acids; Pathogenesis; Pathology; Pathway interactions; Patients; Process; Production; RNA; RNA Editing; Regulation; Reporter; Resolution; Role; Series; Signal Pathway; System; Therapeutic; Work; autoinflammatory; cell type; disease mechanisms study; genome-wide; human disease; in vivo; insight; mortality; mouse model; mutant; novel; nucleic acid detection; postnatal; prevent; pseudotoxoplasmosis syndrome; response; sensor; tool

Project Summary/Abstract Intracellular sensors of nucleic acids are essential for host defense, but they can also cause specific human autoimmune and autoinflammatory diseases if they are not carefully regulated. Over the past 15 years, a paradigm has emerged in which thresholds for activation of these sensors are set by cellular enzymes that metabolize or modify endogenous nucleic acids, thus preventing self-reactivity. The most potent of these enzymes is ADAR1, an RNA editing enzyme that deaminates adenosines in RNA. Mutations in the human ADAR gene that encodes ADAR1 cause Aicardi-Goutieres Syndrome, a severe disease characterized by inappropriate production of type I interferons (IFNs). In previous work, we demonstrated that ADAR1 is an essential and specific regulator of the MDA5-MAVS pathway of antiviral defense. However, the precise relationships between ADAR1 RNA, MDA5 activation, and disease remain incompletely understood because of the lack of tools to study this process in vivo. We have generated three new mouse models to advance our understanding of ADAR1 biology and the broader regulation of the type I IFN response: a knockin mouse model of the most common ADAR AGS allele in humans, a MDA5-HA epitope tag knockin mouse to track MDA5 activation in live cells, and a new Ifnb knockin reporter mouse to enable screens for novel regulators of the type I IFN response. We will use these tools to define the mechanisms that underlie ADAR1-controlled disease in vivo, the precise relationship between ADAR1 RNA editing and MDA5 activation, and the scope of IFN regulation in primary cells. Our studies will uncover fundamental new insights into ADAR1 biology and will identify novel targets that can be manipulated for therapeutic benefit, with direct relevance to human disease.

项目概要/摘要 细胞内核酸传感器对于宿主防御至关重要，但它们也可能导致特定的人类 如果不仔细调节，就会发生自身免疫性疾病和自身炎症性疾病。在过去的 15 年里， 已经出现了这样的范例，其中这些传感器的激活阈值是由细胞酶设定的 代谢或修饰内源核酸，从而防止自身反应。其中最有力的 ADAR1是一种RNA编辑酶，可以使RNA中的腺苷脱氨。人类的突变 编码 ADAR1 的 ADAR 基因会导致 Aicardi-Goutieres 综合征，这是一种严重疾病，其特征为 I 型干扰素 (IFN) 的生产不当。在之前的工作中，我们证明了 ADAR1 是一个 抗病毒防御 MDA5-MAVS 途径的重要和特异性调节剂。然而，准确的 ADAR1 RNA、MDA5 激活和疾病之间的关系仍不完全清楚，因为 缺乏在体内研究这一过程的工具。我们已经生成了三种新的鼠标模型来推进我们的 了解 ADAR1 生物学和 I 型 IFN 反应的更广泛调节：敲入小鼠 人类最常见的 ADAR AGS 等位基因模型，MDA5-HA 表位标签敲入小鼠进行追踪 活细胞中的 MDA5 激活，以及一种新的 Ifnb 敲入报告小鼠，可用于筛选新型调节因子 I 型干扰素反应。我们将使用这些工具来定义 ADAR1 控制的机制 体内疾病、ADAR1 RNA 编辑和 MDA5 激活之间的精确关系以及范围 原代细胞中的干扰素调节。我们的研究将揭示 ADAR1 生物学的基本新见解，并将 确定可操纵以获得治疗效果的新靶点，这些靶点与人类疾病直接相关。