Role of ADAM9 in viral RNA sensing and antiviral innate immunity

ADAM9 在病毒 RNA 传感和抗病毒先天免疫中的作用

基本信息

批准号：
10753041
负责人：
Michaela Ulrike Gack
金额：
$ 26.52万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-02 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10753041
关键词：
2019-nCoV Adaptor Signaling Protein Adult Affect Animal Model Antiviral Response Binding Biochemical Biological Biological Models Brain Cardiac Cell Nucleus Cell surface Cells Characteristics Chemosensitization Complex Coronavirus Coronavirus Infections Coxsackie Viruses Cytoplasmic Tail Cytosol Data Disintegrins Encephalitis Encephalomyocarditis virus Family Family Picornaviridae Filament Genes Genetic Transcription Grant Heart Human IRF3 gene Immune Immune system Immunologic Receptors Impairment Infection Inflammation Inflammatory Influenza A virus Innate Immune Response Integration Host Factors Interferon Activation Interferon Type I Interferon-beta Interferons Knock-out Knockout Mice Knowledge Lead Lentivirus Vector Link Mass Spectrum Analysis Measures Mediating Metalloproteases Mitochondria Mitochondrial Proteins Molecular Morbidity - disease rate Mus Myocarditis Natural Immunity Nucleic Acids Pathogenesis Pathway interactions Picornaviridae Infections Post-Translational Protein Processing Process Production Protein Dephosphorylation Protein Family Proteins RNA RNA Binding RNA Virus Infections RNA Viruses Regulation Research Personnel Role SH3 Domains STAT1 gene Sendai virus Series Signal Pathway Signal Transduction Surface TBK1 gene Tissues Tretinoin Ubiquitination Vesicular stomatitis Indiana virus Viral Viral Encephalitis Viral Genome Viral Pathogenesis Virus Virus Diseases Virus Replication Western Blotting Wild Type Mouse cell type cytosolic receptor detection platform experimental study helicase in vivo innate immune sensing knock-down melanoma member mortality mutant novel prevent protein activation protein protein interaction receptor reconstitution recruit response sensor single-cell RNA sequencing transcriptome sequencing viral RNA viral myocarditis

2019-nCoV Adaptor Signaling Protein Adult Affect Animal Model Antiviral Response Binding Biochemical Biological Biological Models Brain Cardiac Cell Nucleus Cell surface Cells Characteristics Chemosensitization Complex Coronavirus Coronavirus Infections Coxsackie Viruses Cytoplasmic Tail Cytosol Data Disintegrins Encephalitis Encephalomyocarditis virus Family Family Picornaviridae Filament Genes Genetic Transcription Grant Heart Human IRF3 gene Immune Immune system Immunologic Receptors Impairment Infection Inflammation Inflammatory Influenza A virus Innate Immune Response Integration Host Factors Interferon Activation Interferon Type I Interferon-beta Interferons Knock-out Knockout Mice Knowledge Lead Lentivirus Vector Link Mass Spectrum Analysis Measures Mediating Metalloproteases Mitochondria Mitochondrial Proteins Molecular Morbidity - disease rate Mus Myocarditis Natural Immunity Nucleic Acids Pathogenesis Pathway interactions Picornaviridae Infections Post-Translational Protein Processing Process Production Protein Dephosphorylation Protein Family Proteins RNA RNA Binding RNA Virus Infections RNA Viruses Regulation Research Personnel Role SH3 Domains STAT1 gene Sendai virus Series Signal Pathway Signal Transduction Surface TBK1 gene Tissues Tretinoin Ubiquitination Vesicular stomatitis Indiana virus Viral Viral Encephalitis Viral Genome Viral Pathogenesis Virus Virus Diseases Virus Replication Western Blotting Wild Type Mouse cell type cytosolic receptor detection platform experimental study helicase in vivo innate immune sensing knock-down melanoma member mortality mutant novel prevent protein activation protein protein interaction receptor reconstitution recruit response sensor single-cell RNA sequencing transcriptome sequencing viral RNA viral myocarditis

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项目摘要

Project Abstract Picornavirus infections are a leading cause of viral encephalitis and myocarditis in humans. These viral infections can cause substantial inflammatory changes in the brain and heart and lead to significant morbidity and mortality. The immune system detects picornavirus infections, and other single-stranded RNA (ssRNA) viruses, via DEAD/H-box (DDX) helicases that sense cytosolic viral ssRNA and initiate the protective interferon (IFN) response. Two DDX helicases critical in this process are retinoic acid-inducible gene I (RIG-I) and melanoma differentiation-associated gene 5 (MDA5). Both RIG-I and MDA5 are members of the RIG-I-like receptor (RLR) family of RNA-sensing helicases. RIG-I is known to sense negative-sense ssRNA (–ssRNA) viruses (e.g., Sendai virus or vesicular stomatitis virus), while MDA5 is the major sensor for RNA from picornaviruses and other positive-sense ssRNA (+ssRNA) viruses including SARS-CoV2 and other coronaviruses. How MDA5 is activated and regulated is currently not well known, which is in striking contrast to RIG-I for which the activating mechanisms have been elucidated in detail. To study the pathogenesis of myocarditis and encephalitis, researchers have used a prototypical member of the picornavirus family, encephalomyocarditis virus (EMCV). Using EMCV, we demonstrated an important role for the A disintegrin and metalloproteinase protein (ADAM9) in viral pathogenesis. We discovered that mice lacking ADAM9 rapidly succumb to EMCV infection without mounting the characteristic IFN response seen in wild-type mice. Our data indicate a novel role for ADAM9 in viral RNA-induced IFN production through the DDX helicase MDA5. In EMCV infection, MDA5 is the sensor that recognizes viral RNA and initiates a signaling cascade that leads to activation of the mitochondrial antiviral signaling (MAVS) pathway and subsequent IFN production. IFN production is triggered when the viral genome interacts with nucleic acid sensors in the host cell to activate downstream pathways. The host IFN response is crucial to protect the host by limiting virus replication. We hypothesize that ADAM9’s role in viral-induced IFN production is mediated through the MDA5-MAVS pathway. The role of ADAM9 in regulating the IFN response to diverse RNA viruses, including picornaviruses and coronaviruses, will be defined in terms of its effects on the ability of MDA5 to interact with viral RNA and its effect on protein-protein interactions and post-translational modifications of MAVS pathway adapters and effector proteins. Through these experiments, we will define new pathways of IFN activation and better define the pathogenesis of RNA viruses in an animal model system for studying encephalitis and myocarditis.

项目摘要 PICORNAVIRUS感染是人类病毒性脑炎和心肌炎的主要原因。这些病毒感染会引起大脑和心脏的重大炎症变化，并导致明显的发病率和死亡率。免疫系统检测到Picornavirus感染和其他单链RNA（SSRNA）病毒，通过死亡/H-box（DDX）解旋酶，感知胞质病毒ssRNA并启动受保护的干扰素（IFN）响应。在此过程中至关重要的两个DDX解旋酶是视黄酸诱导基因I（RIG-I）和黑色素瘤分化相关基因5（MDA5）。 rig-i和mda5都是类似rig-i的成员 RNA感应解酶的受体（RLR）家族。众所周知，rig-i会感觉到负义ssRNA（–ssRNA）病毒（例如，仙台病毒或囊泡性气孔病毒），而MDA5是RNA的主要传感器 Picornaviruses和其他阳性ssRNA（+SSRNA）病毒，包括SARS-COV2和其他冠状病毒。 MDA5如何激活和调节目前尚不清楚，这与形成鲜明对比 RIG-1已详细阐明了激活机制。为了研究心肌炎和脑炎的发病机理，研究人员使用了典型 Picornavirus家族的成员，脑膜炎病毒（EMCV）。使用EMCV，我们证明了一个 A崩解蛋白和金属蛋白酶蛋白（ADAM9）在病毒发病机理中的重要作用。我们发现缺乏ADAM9的小鼠迅速屈服于EMCV感染而没有安装特征在野生型小鼠中看到的IFN反应。我们的数据表明ADAM9在病毒RNA诱导的IFN中起着新的作用通过DDX解旋酶MDA5生产。在EMCV感染中，MDA5是识别病毒RNA并启动信号级联的传感器导致线粒体抗病毒信号传导（MAV）途径的激活以及随后的IFN产生。当病毒基因组与宿主细胞中的核酸传感器相互作用以激活时，触发了IFN产生下游路径。宿主IFN响应对于通过限制病毒复制来保护宿主至关重要。我们假设ADAM9在病毒诱导的IFN产生中的作用是通过MDA5-MAVS途径介导的。 ADAM9在控制IFN对潜水RNA病毒的反应中的作用，包括Picornavirus和冠状病毒将根据其对MDA5与病毒RNA相互作用及其相互作用的影响而定义对MAVS途径适配器的蛋白质蛋白质相互作用和翻译后修饰的影响效应蛋白。通过这些实验，我们将定义IFN激活的新途径并更好地定义用于研究脑炎和心肌炎的动物模型系统中RNA病毒的发病机理。