NOD2 promotes coxsackievirus entry and pathogenesis

NOD2促进柯萨奇病毒进入和发病机制

基本信息

批准号：
10407068
负责人：
Arina Marijke Keestra-Gounder
金额：
$ 19.44万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-18 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10407068
关键词：
Actins Address Affect Area Binding Cell Culture Techniques Cell Lineage Cell membrane Cells Central Nervous System Diseases Coxsackie Viruses Coxsackievirus Infections Crohn&apos s disease Cytomegalovirus Cytoskeleton Cytosol Data Development Disease Dynamin Endocytosis Endoplasmic Reticulum Endosomes Enteral Enterovirus Enterovirus 71 Epithelial Cells Family Picornaviridae Feces Foundations Future Gastrointestinal tract structure Heart Human Human poliovirus Immunologic Receptors In Vitro Infection Inflammatory Bowel Diseases Inflammatory Response Influenza A virus Insulin-Dependent Diabetes Mellitus Interferon-alpha Intestines Knock-out Knowledge Lead Mediating Medical Membrane Membrane Proteins Mitogen-Activated Protein Kinases Modeling Mus Mycoses Myelogenous Myocarditis Nuclear Nucleotides Oral Organ Pancreas Parasitic infection Pathogenesis Pathway interactions Pattern recognition receptor Peptidoglycan Phosphotransferases Proteins Publishing RIPK2 gene Reporting Research Resistance Respiratory syncytial virus Rhinovirus Role Route Signal Pathway Signal Transduction Site Surface Testing Therapeutic Intervention Tissues Vesicular stomatitis Indiana virus Viral Virulence Virus Virus Diseases Virus Receptors Virus Replication Work adenovirus receptor commensal microbes endoplasmic reticulum stress enteric pathogen experimental study gastrointestinal infection in vivo intestinal epithelium intraperitoneal loss of function mutation mouse model novel nucleotide receptor oral infection pathogen protein function receptor recruit rho GTP-Binding Proteins therapeutic development uptake

Actins Address Affect Area Binding Cell Culture Techniques Cell Lineage Cell membrane Cells Central Nervous System Diseases Coxsackie Viruses Coxsackievirus Infections Crohn&apos s disease Cytomegalovirus Cytoskeleton Cytosol Data Development Disease Dynamin Endocytosis Endoplasmic Reticulum Endosomes Enteral Enterovirus Enterovirus 71 Epithelial Cells Family Picornaviridae Feces Foundations Future Gastrointestinal tract structure Heart Human Human poliovirus Immunologic Receptors In Vitro Infection Inflammatory Bowel Diseases Inflammatory Response Influenza A virus Insulin-Dependent Diabetes Mellitus Interferon-alpha Intestines Knock-out Knowledge Lead Mediating Medical Membrane Membrane Proteins Mitogen-Activated Protein Kinases Modeling Mus Mycoses Myelogenous Myocarditis Nuclear Nucleotides Oral Organ Pancreas Parasitic infection Pathogenesis Pathway interactions Pattern recognition receptor Peptidoglycan Phosphotransferases Proteins Publishing RIPK2 gene Reporting Research Resistance Respiratory syncytial virus Rhinovirus Role Route Signal Pathway Signal Transduction Site Surface Testing Therapeutic Intervention Tissues Vesicular stomatitis Indiana virus Viral Virulence Virus Virus Diseases Virus Receptors Virus Replication Work adenovirus receptor commensal microbes endoplasmic reticulum stress enteric pathogen experimental study gastrointestinal infection in vivo intestinal epithelium intraperitoneal loss of function mutation mouse model novel nucleotide receptor oral infection pathogen protein function receptor recruit rho GTP-Binding Proteins therapeutic development uptake

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项目摘要

Project Summary NOD2 is a cytosolic pattern recognition receptor that senses and responds to bacterial peptidoglycan, activated small Rho GTPases, endoplasmic reticulum stress and viral, fungal and parasitic infections. NOD2 is localized in the cytosol, and upon activation, associates with the plasma membrane and endosomes, which is essential for NOD2 function. Activation of NOD2 results in recruitment of RIPK2 leading to a pro-inflammatory response dependent on NF-B and MAPK signaling pathways. NOD2 restriction of bacterial invasion at the intestinal epithelial surface has been strongly implicated in inflammatory bowel diseases such as Crohn's disease (CD) as loss-of-function mutations in Nod2 can promote the development and onset of CD. Moreover, NOD2 is broadly antiviral as it restricts cytomegalovirus, respiratory syncytial virus, vesicular stomatitis virus and influenza A virus infections. Paradoxically, NOD2 promoted coxsackievirus B3 (CVB3) infection in mice. We found that NOD2 and RIPK2 enhance CVB3 entry in vitro, and NOD2 increases CVB3 pathogenesis in vivo. The following are unknown regarding the impact of NOD2 on CVB3 infection: (1) the mechanism underlying the requirement of NOD2 for efficient CVB3 replication, (2) the novel function for NOD2 during CVB3 entry and (3) the effect of NOD2 on CVB3 infection in the gastrointestinal tract, the initial and natural site of infection. We hypothesize that CVB3 has evolved to usurp NOD2 for optimal viral replication in the gastrointestinal tract to promote CVB3 dissemination to other organs, which causes increased CVB3- induced pathogenesis. We will test our hypothesis by pursuing two aims. For Aim 1, we will use in vitro cell culture infection models in intestinal epithelial cells (IEC) to determine the mechanism for NOD2 enhancement of CVB3 entry into cells. We will investigate whether NOD2 impacts CVB3 binding to its receptor (CAR), if NOD2 is recruited to sites of CVB3 entry and whether membrane localization is required for CVB3 to enter cells. We will also assess whether NOD2 affects CVB3 uptake by influencing the cell cytoskeleton. Aim 2 will examine the in vivo contribution of NOD2 on CVB3 infection. We will identify what cell lineage is critical for the impact of NOD2 on CVB3 infection and pathogenesis in a mouse model. As CVB3 is a fecal-oral transmitted virus that infects the intestine prior to spreading to other organs, we will use an oral infection model in our Nod2-sufficient and -deficient mice, using an Interferon alpha/beta receptor knock-out (Ifnar1-/-) background to render mice more orally susceptible, to ascertain the importance of NOD2 during intestinal CVB3 infection. Altogether, these studies will define how CVB3 exploits the innate immune receptor NOD2. Information from the proposed research could lead to the development of therapeutics for coxsackievirus infections and possibly other enteroviruses, will help us understand host intestinal influences on CVB3 and will propel future studies focused on other viral infections.

项目摘要 NOD2是一种胞质模式识别受体，感知并响应细菌肽聚糖，激活的小鼠体，内质网应激以及病毒，真菌和寄生虫感染。 nod2是局部在细胞质中，激活后，与质膜和内体相关联， NOD2功能必不可少的。 NOD2的激活导致RIPK2的募集导致促炎性响应取决于NF-B和MAPK信号通路。 NOD2限制细菌入侵肠上皮表面在炎症性肠病（例如克罗恩）中强烈暗示疾病（CD）作为NOD2的功能丧失突变可以促进CD的发育和发作。而且， NOD2是巨大的抗病毒，因为它限制了巨细胞病毒，呼吸综合病毒，囊泡气孔病毒病毒和影响病毒感染。矛盾的是，NOD2在小鼠中促进了Coxsackievievirus B3（CVB3）感染。我们发现NOD2和RIPK2在体外增强了CVB3进入，NOD2增加了CVB3发病机理体内。关于NOD2对CVB3感染的影响尚不清楚：（1）机制 NOD2对有效CVB3复制的需求，（2）NOD2的新功能 CVB3进入和（3）NOD2对胃肠道的CVB3感染的影响，初始和自然部位感染。我们假设CVB3已演变为篡夺NOD2，以在最佳的病毒中复制胃肠道以促进CVB3向其他器官传播，这导致CVB3-- 诱导的发病机理。我们将通过追求两个目标来检验我们的假设。对于AIM 1，我们将使用体外细胞肠上皮细胞（IEC）中的培养感染模型，以确定NOD2增强的机制 CVB3进入细胞。我们将调查NOD2是否影响CVB3与其接收器（CAR）结合（是否） NOD2被招募到CVB3条目的站点，以及CVB3是否需要膜定位细胞。我们还将评估NOD2是否通过影响细胞骨架来影响CVB3的摄取。 AIM 2意志检查NOD2对CVB3感染的体内贡献。我们将确定哪种细胞谱系对 NOD2对小鼠模型中CVB3感染和发病机理的影响。因为CVB3是一种粪便的传播在传播到其他器官之前感染肠道的病毒，我们将在我们的口腔感染模型中使用使用干扰素α/beta受体敲除（IFNAR1 - / - ）背景为NOD2充足和缺陷小鼠使小鼠口服更容易受到影响，以确定肠道CVB3感染期间NOD2的重要性。总的来说，这些研究将定义CVB3如何利用先天免疫受体NOD2。来自拟议的研究可能会导致Coxsackievivirus感染理论的发展，并可能其他肠病毒将帮助我们了解宿主肠道对CVB3的影响，并将推动未来的研究专注于其他病毒感染。