Role of microRNAs in Coronavirus-induced lethal encephalitis

microRNA 在冠状病毒引起的致死性脑炎中的作用

• 批准号: 8190928
• 负责人: SONIA NAVAS-MARTIN
• 金额: $ 23.17万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8190928
• 关键词: 3' Untranslated Regions; Acute; Animal Model; Binding; Biological; Brain; CD8B1 gene; Cell physiology; Central Nervous System Diseases; Chronic; Computer Simulation; Coronavirus; Coronavirus Infections; Data; Disease; Disease Outcome; Encephalitis; Encephalomyelitis; Etiology; Experimental Models; Functional RNA; Gene Expression; Genetic Transcription; Human; Immune response; Immune system; In Vitro; Infection; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Interferon Type II; Interferon-beta; Interferons; Interleukin-1; Investigation; Lead; Lentivirus Vector; Leukocytes; Mediating; Meningoencephalitis; Messenger RNA; MicroRNAs; Microarray Analysis; Microglia; Molecular Profiling; Murine hepatitis virus; Mus; Natural Immunity; Neuraxis; Neuroglia; Neuropathogenesis; Neutrophil Infiltration; Outcome; Pathogenesis; Pathway interactions; Pattern; Play; Poly(A)+ RNA; Population; RNA Viruses; Regulation; Relative (related person); Reverse Transcriptase Polymerase Chain Reaction; Role; Severities; Small RNA; Subfamily lentivirinae; T-Lymphocyte; Time; Transcript; Translational Repression; Untranslated Regions; Validation; Viral; Viral Encephalitis; Virus; Virus Diseases; anakinra; base; chemokine; chronic demyelination; cytokine; in vivo; knock-down; mRNA Expression; mRNA Transcript Degradation; macrophage; neuroprotection; neurovirulence; novel; novel therapeutic intervention; pathogen; prevent; receptor; response; therapeutic development; viral RNA; virus host interaction; white matter

DESCRIPTION (provided by applicant): MicroRNAs (miRNAs) are small RNAs that regulate gene expression by translational repression and/or mRNA degradation through binding to the 3'UTR of target mRNAs. There is increasing evidence indicating that miRNAs play a major role in fundamental cellular processes, the regulation of the immune system, as well as in the onset and progression of many diseases. Although the role of miRNAs in virus-host interactions is emerging, there is little evidence that viral modulation of cellular miRNAs actually impacts neuro-inflammatory responses in vivo. Unraveling the roles of miRNAs in viral encephalitis in humans is challenging. The study of animal models of viral encephalitis is key to our understanding of how viruses drive disease in the CNS. Differences in the severity of viral encephalitis may reflect the differential ability of viruses to stimulate innate immune responses within the CNS. Coronaviruses (CoVs) are enveloped, positive-sense, single-stranded, polyadenylated RNA viruses. Different murine CoVs (Mouse Hepatitis Virus, MHV) isolates induce acute lethal encephalitis and encephalomyelitis associated with acute and chronic demyelination. Here, we will compare two strains differing markedly in their neurovirulence: MHV-JHM, a highly neurovirulent strain that produces lethal encephalitis; and MHV-A59, a mildly neurovirulent strain that induces acute meningoencephalitis and chronic. In preliminary studies, we have used miRNA microarray analysis and validation by real-time RT-PCR to provide evidence of the differential expression of selected cellular miRNAs in macrophages upon in vitro infection with MHV-A59 and MHV-JHM. Changes in the macrophage miRNAs profiles after murine CoVs infection were overlapping but distinct, and were observed early after infection in the absence of type I IFNs, IFN-?, and pro-inflammatory cytokine secretion. Our data suggest that MHV infection can modify the pattern of cellular miRNA expression. We have identified two miRNAs that are significantly up-regulated by the highly neurovirulent MHV-JHM (but not by MHV-A59). Interestingly, we also found that these JHM-up-regulated miRNAs potentially target the 3' UTR of IL-1 receptor antagonist (IL-1Ra), a cytokine involved in both neuroprotection and neuropathogenesis, as well as IFN?. We hypothesize that CoVs inducing different CNS disease outcome promote distinct miRNAs expression profiles in macrophages and microglia, and that virus specific changes in miRNAs expression contribute to the differences in encephalitis outcome by modulation of the inflammatory response. We propose to: 1) systematically identify miRNAs and mRNAs regulatory networks that correlate with lethal encephalitis progression in the infected brain; 2) Define the role of those JHM-up-regulated miRNAs (identified in preliminary studies and in Specific Aim 1) in IL-1Ra and IFN? induction by macrophages and glial cells ex vivo, as well as in vivo using antagomir-lentivirus vectors. Overall, there is significant translational potential in the proposed investigations since they will facilitate the development of therapeutic approaches to prevent and/or treat neuro-inflammatory diseases of viral etiology. PUBLIC HEALTH RELEVANCE: The relative contribution of microRNAs to virus-host interactions in the context of the CNS, and to neuroinflammatory responses and/or neuropathogenesis is poorly understood. Our experimental model will provide a distinctive opportunity to define the role of selected microRNAs in neuro-inflammatory responses that result in viral-induced lethal encephalitis, and could potentially lead to novel therapeutic approaches to prevent and/or treat acute neuro-inflammatory diseases of viral etiology.

描述（由申请人提供）：microRNA（miRNA）是小的RNA，通过翻译抑制和/或mRNA降解通过与靶标mRNA的3'UTR结合来调节基因表达。越来越多的证据表明，miRNA在基本细胞过程，免疫系统的调节以及许多疾病的发作和进展中起着重要作用。尽管miRNA在病毒宿主相互作用中的作用正在出现，但几乎没有证据表明细胞miRNA的病毒调节实际上会影响体内神经炎症反应。揭开miRNA在人类病毒脑炎中的作用是具有挑战性的。对病毒脑炎动物模型的研究是我们了解病毒如何驱动中枢神经系统疾病的关键。病毒脑炎严重程度的差异可能反映了病毒刺激中枢神经系统中先天免疫反应的差异能力。冠状病毒（COV）被包裹，阳性，单链，聚腺苷酸化的RNA病毒。不同的鼠COV（小鼠肝炎病毒，MHV）分离出与急性和慢性脱髓鞘有关的急性致死性脑炎和脑脊髓炎。在这里，我们将比较两种菌株的神经动力毒气：MHV-JHM，一种高度神经性肺部菌株，会产生致命的脑炎；和MHV-A59，一种轻度神经性肺部菌株，可诱导急性脑膜脑炎和慢性。在初步研究中，我们已经使用miRNA微阵列分析和实时RT-PCR验证来提供巨噬细胞中选定细胞miRNA在巨噬细胞中的差异表达的证据，该表达在体外感染了MHV-A59和MHV-JHM后。鼠Covs感染后巨噬细胞miRNA轮廓的变化是重叠但截然不同的，并且在没有I型IFN，IFN-？和促炎性细胞因子分泌的情况下感染了早期。我们的数据表明，MHV感染可以改变细胞miRNA表达的模式。我们已经确定了两种由高度神经性肺部MHV-JHM显着上调的miRNA（但不是由MHV-A59）。有趣的是，我们还发现，这些JHM-UP调节的miRNA可能靶向IL-1受体拮抗剂（IL-1RA）的3'UTR，这是一种参与神经保护和神经病变的细胞因子，以及IFN？我们假设诱导不同中枢神经系统疾病结果的COV促进了巨噬细胞和小胶质细胞中不同的miRNA表达谱，而miRNAS表达的病毒特异性变化通过调节炎症反应的调节导致脑炎结局的差异。我们建议：1）系统地识别与感染大脑中致死性脑炎进展相关的miRNA和mRNA调节网络； 2）在IL-1RA和IFN中定义了那些JHM-UP调节的miRNA（在初步研究和特定目标中鉴定）的作用？巨噬细胞和神经胶质细胞的诱导，以及使用Antagomir-Lentivirus载体的体内诱导。总体而言，拟议的研究中具有巨大的转化潜力，因为它们将促进预防和/或治疗病毒病因的神经炎症性疾病的治疗方法的发展。 公共卫生相关性：在中枢神经系统的背景下，microRNAS对病毒宿主相互作用的相对贡献，以及神经炎症反应和/或神经发病的相对贡献。我们的实验模型将提供一个独特的机会，以定义选定的microRNA在神经炎症反应中的作用，从而导致病毒引起的致死性脑炎，并有可能导致预防和/或治疗病毒病理学的新型治疗方法来预防和/或治疗急性神经炎性疾病。