NOD2 promotes coxsackievirus entry and pathogenesis

NOD2促进柯萨奇病毒进入和发病机制

• 批准号: 10285845
• 负责人: Arina Marijke Keestra-Gounder
• 金额: $ 23.33万
• 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 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10285845
• 关键词: Actins; Address; Affect; Antiviral Agents; 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 NOD 2是一种细胞溶质模式识别受体，可感知并响应细菌肽聚糖， 活化的小Rho GTP酶、内质网应激和病毒、真菌和寄生虫感染。NOD 2是 定位于胞质溶胶中，并且在活化时，与质膜和内体缔合， NOD 2的功能。NOD 2的激活导致RIPK 2的募集，导致促炎性细胞因子的产生。 反应依赖于NF-κ B B和MAPK信号通路。NOD 2限制细菌侵入 肠上皮表面与炎症性肠病如克罗恩病密切相关 Nod 2的功能缺失突变可促进CD的发展和发作。此外，委员会认为， NOD 2是广泛的抗病毒，因为它限制巨细胞病毒，呼吸道合胞病毒，水泡性口炎病毒， 和甲型流感病毒感染。NOD 2可促进小鼠柯萨奇病毒B3（CVB 3）感染。 我们发现NOD 2和RIPK 2在体外增强了CVB 3的进入，并且NOD 2在体外增加了CVB 3的致病性。 vivo.关于NOD 2对CVB 3感染的影响，以下是未知的：（1）机制 NOD 2对CVB 3有效复制的潜在需求，（2）NOD 2在CVB 3复制过程中的新功能， NOD 2对CVB 3在胃肠道中感染的影响，即初始和自然部位 感染我们假设，CVB 3已经进化到取代NOD 2，以实现最佳的病毒复制。 胃肠道促进CVB 3传播到其他器官，这导致CVB 3- 诱发发病机制。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 在肠上皮细胞（IEC）中培养感染模型以确定NOD 2增强的机制 CVB 3进入细胞。我们将研究NOD 2是否影响CVB 3与其受体（CAR）的结合，如果 NOD 2被募集到CVB 3进入的位点，以及CVB 3进入是否需要膜定位 细胞我们还将评估NOD 2是否通过影响细胞骨架来影响CVB 3摄取。目标2将 检测NOD 2对CVB 3感染的体内作用。我们将确定什么样的细胞谱系对于 小鼠模型中NOD 2对CVB 3感染和发病机制的影响。由于CVB 3是一种粪-口传播的病毒， 在传播到其他器官之前感染肠道的病毒，我们将在我们的研究中使用口腔感染模型。 使用干扰素α/β受体敲除（Ifnar 1-/-）背景， 使小鼠更加口服敏感，以确定NOD 2在肠道CVB 3感染期间的重要性。 总之，这些研究将确定CVB 3如何利用先天免疫受体NOD 2。信息从 这项拟议中的研究可能会导致柯萨奇病毒感染的治疗方法的发展， 其他肠道病毒，将有助于我们了解宿主肠道对CVB 3的影响，并将推动未来的研究 专注于其他病毒感染