NOD2 promotes coxsackievirus entry and pathogenesis

NOD2促进柯萨奇病毒进入和发病机制

• 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
• 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' 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

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，可以感知细菌的肽聚糖并对其做出反应， 激活的小Rho GTP酶、内质网应激以及病毒、真菌和寄生虫感染。NOD2是 定位在胞浆中，激活后与质膜和内小体结合，这是 对于NOD2功能来说是必不可少的。NOD2的激活导致RIPK2的募集，从而导致促炎 反应依赖于NF-B和MAPK信号通路。NOD2对细菌入侵的限制 肠道上皮细胞表面与炎症性肠病(如克罗恩病)密切相关 疾病(CD)是指NOD2功能缺失突变，可促进CD的发生发展。此外， NOD2对巨细胞病毒、呼吸道合胞病毒、水泡性口炎病毒具有广泛的抗病毒作用 和甲型流感病毒感染。矛盾的是，NOD2促进了柯萨奇病毒B3(CVB3)在小鼠体内的感染。 我们发现，NOD2和RIPK2在体外促进CVB3进入，而NOD2促进CVB3在体外的致病作用。 活着。以下是关于NOD2对CVB3感染的影响的未知之处：(1)机制 基于NOD2对高效CVB3复制的需求，(2)NOD2在 CVB3进入和(3)NOD2对CVB3在胃肠道、起始部位和自然部位感染的影响 感染的可能性。我们假设CVB3已经进化为篡夺NOD2，以便在 胃肠道促进CVB3扩散到其他器官，从而导致CVB3- 诱导发病机制。我们将通过追求两个目标来检验我们的假设。对于目标1，我们将使用体外细胞 肠上皮细胞(IEC)培养感染模型研究NOD2增强机制 CVB3进入细胞的比例。我们将调查NOD2是否影响CVB3与其受体(CAR)的结合，如果 NOD2被招募到CVB3进入的部位以及是否需要膜定位才能进入CVB3 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。目标2将 检测NOD2在体内对CVB3感染的贡献。我们将确定什么细胞谱系对 NOD2在小鼠模型中对CVB3感染和发病机制的影响。由于CVB3是一种粪便-口腔传播 在传播到其他器官之前先感染肠道的病毒，我们将在我们的 使用干扰素α/β受体敲除(Ifnar1-/-)背景的NOD2充足和缺陷小鼠 使小鼠对口服更敏感，以确定NOD2在肠道CVB3感染中的重要性。 总之，这些研究将确定CVB3如何利用先天免疫受体NOD2。信息来自 这项拟议的研究可能会导致柯萨奇病毒感染的治疗方法的发展，并可能 其他肠道病毒，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。