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.

项目概要 NOD2 是一种胞质模式识别受体，可感知细菌肽聚糖并对其做出反应， 激活小 Rho GTP 酶、内质网应激以及病毒、真菌和寄生虫感染。 NOD2 是 定位于细胞质中，并在激活后与质膜和内体结合，这是 对于 NOD2 功能至关重要。 NOD2 的激活导致 RIPK2 的募集，从而导致促炎 反应依赖于 NF-κB 和 MAPK 信号通路。 NOD2 限制细菌入侵 肠上皮表面与克罗恩病等炎症性肠病密切相关 疾病（CD）作为Nod2的功能丧失突变可以促进CD的发展和发病。而且， NOD2 具有广泛的抗病毒作用，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。矛盾的是，NOD2 促进小鼠柯萨奇病毒 B3 (CVB3) 感染。 我们发现NOD2和RIPK2在体外增强CVB3进入，并且NOD2在体内增强CVB3发病机制 体内。关于NOD2对CVB3感染的影响尚不清楚：（1）机制 NOD2 对 CVB3 有效复制的要求的基础，(2) NOD2 在复制过程中的新功能 CVB3进入和（3）NOD2对胃肠道中CVB3感染的初始和自然位点的影响 的感染。我们假设 CVB3 已经进化为取代 NOD2 以实现最佳病毒复制 胃肠道促进 CVB3 传播到其他器官，从而导致 CVB3- 增加 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 肠上皮细胞 (IEC) 培养感染模型以确定 NOD2 增强的机制 CVB3 进入细胞的过程。我们将研究 NOD2 是否影响 CVB3 与其受体 (CAR) 的结合，如果 NOD2 被招募到 CVB3 进入位点以及 CVB3 进入是否需要膜定位 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。目标2将 检查 NOD2 对 CVB3 感染的体内贡献。我们将确定哪些细胞谱系对于 NOD2 对小鼠模型中 CVB3 感染和发病机制的影响。由于 CVB3 是粪口传播的 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 Nod2 充足和缺乏的小鼠，使用干扰素 α/β 受体敲除 (Ifnar1-/-) 背景 使小鼠更容易经口敏感，以确定 NOD2 在肠道 CVB3 感染过程中的重要性。 总而言之，这些研究将明确 CVB3 如何利用先天免疫受体 NOD2。信息来自 拟议的研究可能会导致柯萨奇病毒感染疗法的开发，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 重点关注其他病毒感染。