Link between Retroelements, Ro and Interferon Biology in Lupus

狼疮中逆转录元素、Ro 和干扰素生物学之间的联系

• 批准号: 9378686
• 负责人: Keith B. Elkon
• 金额: $ 23.23万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-21 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9378686
• 关键词: Adjuvant; Antigen-Antibody Complex; Attenuated; Autoantibodies; Autoantigens; Autoimmune Diseases; Autoimmunity; B-Lymphocytes; Binding; Biology; Boron; Cell Line; Cell surface; Cells; Clinical; Congenital Heart Block; Cutaneous Lupus Erythematosus; DNA Damage; Data; Dendritic Cells; Disease; Disease susceptibility; Elements; Embryo; Employee Strikes; Environmental Risk Factor; Enzymes; Estrogens; Exanthema; Female; Fibroblasts; Genes; Genetic; Genetic Transcription; Hormones; Human; Immune signaling; Immunoprecipitation; In Vitro; Inbred MRL lpr Mice; Inflammation; Inflammatory; Interferon Type I; Interferon-beta; Interferons; Knockout Mice; Lead; Learning; Ligands; Link; Lupus; Mediating; Methods; Methylation; Methyltransferase; Mouse Strains; Mus; Mutation; Natural Immunity; Nucleic Acids; Pathogenesis; Pathway interactions; Patients; Photosensitivity; Prevention; Proteins; RNA; RNA Binding; Reporting; Research; Research Personnel; Retroelements; Retrotransposon; Role; SS-A antibodies; Short Interspersed Nucleotide Elements; Signal Pathway; Signal Transduction; Sjogren' s Syndrome; Skin; Stimulus; Systemic Lupus Erythematosus; TREX1 gene; Techniques; Testing; To autoantigen; Transcript; UV induced; Ultraviolet B Radiation; Ultraviolet Rays; Untranslated RNA; Work; base; cytokine; experience; experimental study; in vivo; in vivo Model; insight; knock-down; lupus-like; male; new technology; overexpression; programs; receptor; response; sensor; targeted treatment; therapy design; ultraviolet irradiation

Project Summary A variety of nucleic acid ligands have been implicated in the striking Type I Interferon (IFN-I) response observed in lupus and related diseases but which ligands and receptors drive disease are unclear. Amongst putative ligands are retroelements (RE), one class of which, the SINES, was recently shown to bind to the autoantigen, Ro60. Tying this observation to the surprising finding that Ro60 deficient mice get lupus, we will test an explanation for lupus pathogenesis that links disease susceptibility to Ro function. To test the hypothesis that an excess of SINES relative to Ro induces IFN-I and a lupus like disease, we propose: In Aim 1, we will identify the small non coding RNAs that bind to murine Ro60 and we will define the Ro recognition motifs (RRM) using a highly sensitive and specific immunoprecipitation technique called iCLIP. In addition, we will generate mouse embryonic fibroblasts (MEFS) that are deficient in Ro and in one of the key RNA sensors called RIG-I or MDA5 that stimulate IFN-I signaling. These experiments will identify the RNA ligand, motif(s) and corresponding RNA sensors responsible for cell autonomous IFN-I stimulation. In the second Aim, we will ask how Ro60 and RNA binding elements are regulated, in response to lupus relevant environmental triggers. First, using cell based approaches and the MEFS generated in Aim 1, we will de-repress SINES by inhibiting the methylase that is responsible for suppression of SINE transcription in mice. Next, we will expose cells to UVB radiation that has been shown to result in increased SINE expression. Finally, as prompted by our preliminary data, we will examine whether Estrogen (E2) upregulates SINES or reduces Ro expression. The experiments are expected to validate SINES as the key RNA ligand that provokes cell intrinsic IFN-I stimulation and test whether lupus related environmental factors alter expression of the RNA or protein. Following these experiments, we will examine the same environmental factors in Ro deficient mice. Since Ro deficient mice develop a lupus like autoimmunity for which there is no current explanation, we will explore the hypothesis that, when Ro is limiting or SINES are overexpressed, SINES stimulate IFN-I which in turn leads to a lupus like disease in the appropriate genetic background. We expect that UV light will increase IFN-I expression in the skin and that this finding will be markedly exacerbated in Ro deficient female mice. Anti-Ro antibodies are usually the first autoantibody to develop in human lupus – many years before the onset of clinical disease. They are also associated with congenital heart block and UV mediated skin rashes. Through understanding the roles of SINE elements and Ro in triggering specific IFN pathways in lupus, we should be able to explore the pathobiology with much greater insight in human SLE and devise approaches to targeted prevention and treatment.

项目摘要 多种核酸配体与显著的I型干扰素（IFN-1）应答有关 在狼疮和相关疾病中观察到，但哪种配体和受体驱动疾病尚不清楚。之间 假定的配体是逆向元素（RE），其中一类，西内斯，最近被证明与 自身抗原，Ro 60。将这一观察结果与Ro 60缺陷小鼠患狼疮的惊人发现联系起来，我们将 测试狼疮发病机制的解释，将疾病易感性与Ro功能联系起来。测试 假设相对于Ro过量的西内斯诱导IFN-1和狼疮样疾病，我们提出： 在目标1中，我们将鉴定与鼠Ro 60结合的小的非编码RNA，并且我们将定义Ro 识别基序（RRM），使用称为iCLIP的高灵敏度和特异性免疫沉淀技术。在 此外，我们将产生小鼠胚胎成纤维细胞（MEFS），这是缺乏Ro和在一个关键的 RNA传感器RIG-I或MDA 5刺激IFN-I信号。这些实验将鉴定出 配体、基序和负责细胞自主IFN-1刺激的相应RNA传感器。 在第二个目标中，我们将询问Ro 60和RNA结合元件是如何调节的， 狼疮相关的环境诱因首先，使用基于单元的方法和目标1中生成的MEFS， 我们将通过抑制负责抑制SINE转录的甲基化酶来去抑制西内斯 对小鼠接下来，我们将细胞暴露于UVB辐射，该辐射已被证明会导致SINE增加。 表情最后，根据我们的初步数据，我们将研究雌激素（E2）是否上调 西内斯或减少Ro表达。预计这些实验将验证西内斯作为关键的RNA配体， 激发细胞内在的IFN-Ⅰ刺激，并测试狼疮相关的环境因素是否改变 RNA或蛋白质。在这些实验之后，我们将在Ro中检查相同的环境因素 缺陷小鼠由于Ro缺陷小鼠发展出狼疮样自身免疫， 解释，我们将探讨的假设，当Ro是有限的或西内斯过表达，西内斯 刺激IFN-I，这反过来导致在适当的遗传背景下的狼疮样疾病。我们预计 紫外线会增加皮肤中IFN-I的表达，而这一发现在Ro中会明显加剧。 缺乏雌性小鼠。 抗Ro抗体通常是人类狼疮中产生的第一个自身抗体--很多年前 临床疾病的开始。它们也与先天性心脏传导阻滞和紫外线介导的皮肤有关 疹子通过了解SINE元件和Ro在狼疮中触发特异性IFN途径的作用， 我们应该能够更深入地了解人类SLE的病理生物学， 有针对性的预防和治疗。