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 I型I型干扰素（IFN-I）反应中暗示了多种核酸配体 在狼疮和相关疾病中观察到，但哪些配体和接收器驱动疾病尚不清楚。中间 假定的配体是恢复元素（re），最近证明了一类，罪恶是与罪相结合的 Autoantigen，RO60。将这一观察结果与令人惊讶的发现，RO60缺乏老鼠得到狼疮，我们将 测试狼疮发病机理的解释，该发病机理将疾病易感性与RO功能联系起来。测试 假设相对于RO诱导IFN-I和像狼疮这样的罪恶过多，我们提出： 在AIM 1中，我们将确定与Murine RO60结合的小型非编码RNA，我们将定义RO 使用称为ICLIP的高度敏感和特定的免疫沉淀技术的识别基序（RRM）。在 此外，我们还将生成不足RO和一个键之一的小鼠胚胎成纤维细胞（MEF） RNA传感器称为RIG-I或MDA5，刺激IFN-I信号传导。这些实验将识别RNA 配体，基序（S）和相应的RNA传感器负责细胞自主IFN-I刺激。 在第二个目标中，我们将询问如何调节RO60和RNA结合元件，以响应于 狼疮相关的环境触发器。首先，使用基于单元的方法和AIM 1中生成的MEF， 我们将通过抑制负责抑制正弦转录的甲基化酶来取消抑制罪 在老鼠中。接下来，我们将使细胞暴露于已显示导致正弦增加的UVB辐射 表达。最后，正如我们的初步数据提示的那样，我们将检查雌激素（E2）是否上调 罪或降低RO表达。预计该实验将验证罪恶为关键的RNA配体 引起细胞固有的IFN-I刺激，并测试狼疮相关的环境因素是否改变了 RNA或蛋白质。在这些实验之后，我们将检查RO中相同的环境因素 由于缺乏RO的小鼠会产生像自动免疫这样的狼疮 说明，我们将探讨以下假设：当ro限制或罪过过表达时， 刺激IFN-I，进而导致在适当的遗传背景下类似狼疮的疾病。我们期望 紫外线会增加皮肤中的IFN-I表达，并且该发现将在RO中明显加剧 雌性小鼠不足。 抗RO抗体通常是人类狼疮中第一个开发的自身抗体 - 多年以前 临床疾病的发作。它们还与先天性心脏块和紫外线介导的皮肤有关 皮疹。通过了解正弦元素和RO在触发狼疮中特定的IFN途径中的作用， 我们应该能够在人类的SLE和设计方法中探索病理生物学 进行针对性的预防和治疗。