Intracellular nucleic acid detection in autoimmunity

自身免疫中的细胞内核酸检测

• 批准号: 10088370
• 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/10088370
• 关键词: ADAR1; Acute; Address; Adenosine; Alleles; Antiviral Agents; 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.

项目摘要/摘要 核酸的细胞内传感器对于宿主防御至关重要，但它们也会引起特定的人类 自身免疫性和自身炎症性疾病如果不仔细调节。在过去的15年中， 出现了范式，其中用于激活这些传感器的阈值是由细胞酶设定的 代谢或修饰内源性核酸，从而防止自我反应性。其中最有效的 酶是ADAR1，是一种RNA编辑酶，可脱氨腺苷在RNA中。人类的突变 编码ADAR1的ADAR基因引起Aicardi-Goutieres综合征，这种严重疾病为特征 I型干扰素的不适当产生（IFNS）。在以前的工作中，我们证明了Adar1是 MDA5-MAVS抗病毒防御途径的必不可少的调节剂。但是，确切的 ADAR1 RNA，MDA5激活和疾病之间的关系仍然不完全理解 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 对ADAR1生物学的理解和I型IFN响应的更广泛调节：敲击小鼠 在人类中最常见的Adar AGS等位基因的模型 活细胞中的MDA5激活，以及一种新的IFNB敲蛋白记者小鼠，以启用用于新型调节剂的屏幕 I型IFN响应。我们将使用这些工具来定义基于ADAR1控制的机制 疾病在体内，ADAR1 RNA编辑与MDA5激活之间的精确关系以及 原代细胞中的IFN调节。我们的研究将发现对ADAR1生物学的基本新见解，并将 确定可以通过治疗益处来操纵的新型靶标，并且与人类疾病直接相关。