The role of TRIM2 and SIRPA in New World Arenavirus entry

TRIM2 和 SIRPA 在新世界沙粒病毒进入中的作用

基本信息

批准号：
10625278
负责人：
SUSAN R ROSS
金额：
$ 49.38万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-23 至 2027-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10625278
关键词：
2019-nCoV Actins Aerosols Amino Acid Motifs Antiviral Response Antiviral Therapy Apoptosis Arenavirus Arenavirus Infections Argentina Binding Biological Process Blood Platelets C-terminal Cell Culture Techniques Cell Extracts Cell Proliferation Cell Surface Proteins Cell surface Cells Complex Cultured Cells Cytoskeleton Data Disease Ebola Endocytosis Endothelium Erythrocytes Family Functional disorder Genes Glycoproteins Goals Human Human Pathology Image Immune response Immune system In Vitro Incidence Infection Infection Control Integration Host Factors Junin virus Knockout Mice Link Lymphocytic choriomeningitis virus Macrophage Mediating Modeling Mus Mutation Old World Arenaviruses PTPN6 gene PTPNS1 gene Pathogenicity Pathway interactions Patients Phagocytosis Phagocytosis Inhibition Phosphoric Monoester Hydrolases Phosphorylation Primates Process Protein Family Proteins RNA Viruses RNA interference screen Rodent Role Sentinel Signal Transduction Signal Transduction Pathway Small Interfering RNA Specificity Survivors TFRC gene TRIM Motif Tacaribe Complex Viruses Tacaribe virus Testing Therapeutic Intervention Tissue Extracts Ubiquitination Vaccines Vesicular stomatitis Indiana virus Viral Viral Hemorrhagic Fevers Virus Virus Diseases Visualization Work Zika Virus Zoonoses cell type cellular targeting cytokine experimental study in vivo insight knock-down member mortality neoplastic cell nervous system disorder novel novel strategies novel therapeutic intervention pathogenic virus prevent receptor therapeutic target therapeutically effective trafficking transmission process tumor ubiquitin ligase viral transmission

项目摘要

New world hemorrhagic fever arenaviruses (NWAs), such as Junín virus, are rodent-transmitted viruses that cause ~30% mortality when they zoonose into humans. The mechanism by the NWAs induce disease is still not certain, although it likely includes induction of high levels of cytokines by infected sentinel cells of the immune system, leading to endothelia and thrombocyte dysfunction and neurological disease. Survivors of Junín infection develop strong humoral immune responses, suggesting that controlling infection at early times post-infection is critical for virus clearance. Although an effective Junín virus vaccine has decreased disease incidence, sporadic cases of this as well as the other known and novel NWAs for which there are no vaccines or effective therapeutics still occur. It is well-established that the clade B pathogenic NWAs bind to transferrin receptor 1 and other receptors on the cell surface, but the steps leading to their entry from an acidic cellular compartment are not well-determined. We recently performed a siRNA screen with pseudotyped viruses bearing a pathogenic Junín glycoprotein with the goal of finding host genes involved in entry that could serve as therapeutic targets. We found that TRIM2, a member of the tripartite motif family that includes well-known members of the host's intrinsic defense against viral infections, limits NWA endocytosis into cells. By probing the TRIM2 interactome for other host proteins that block NWA infection, we discovered that SIRPA, a cell surface protein that inhibits macrophage phagocytosis of tumor and dead cells and erythrocytes, also decreases infection. Importantly, SIRPA, unlike TRIM2, inhibits infection by various human pathogenic viruses that require trafficking to an acidic compartment, including VSV, Zika virus, LCMV and Ebola and SARS-Cov-2 pseudoviruses. Our data suggest that TRIM2 and SIRPA act at the viral entry/internalization step. These finding suggests that there are common mechanisms that regulate virus endocytosis and phagocytosis. We propose here to further investigate the overlap between virus-mediated endocytosis and phagocytosis in vitro, ex vivo and in vivo in three aims that will 1) investigate the overlap in the NWA entry and phagocytosis pathways; 2) determine where TRIM2/SIRPA inhibition of infection occurs; and 3) use TRIM2, SIRPA and other relevant knockout mice to probe the roles of these proteins in cell-type specific and in vivo infection by replication-competent NWAs. In addition to providing mechanistic insight into the entry of NWAs into cells, these studies have the potential of increasing our understanding as to how host factors limit infection and could lead to new approaches to therapeutic intervention.

新世界出血热轨迹（NWAS），例如junín病毒，是啮齿动物传播的当缩小人类时，导致约30％死亡率的病毒。 NWAS的机制诱导疾病仍然不确定，尽管它可能包括诱导高水平的细胞因子通过免疫系统的感染前哨细胞，导致内皮细胞和血小板细胞功能障碍和神经疾病。朱恩感染的幸存者发展出强烈的幽默免疫反应，表明感染后感染在感染后对病毒清除。尽管有效的junín病毒疫苗疾病事件减少，但零星的情况以及其他没有阴道的已知和新颖的NWA 或有效的疗法仍在发生。良好的表明，进化枝B致病性NWA结合转运蛋白受体1和细胞表面上的其他受体，但导致其进入的步骤从酸性细胞室中没有明确确定。我们最近进行了siRNA 带有病原性junín糖蛋白的伪型病毒的筛选，目的是查找参与进入的寄主基因，可以用作治疗靶标。我们发现Trim2，一个 Tripite Motif家族的成员，其中包括主持人的内在成员防御病毒感染，将NWA内吞作用限制为细胞。通过探测trim2 用于阻断NWA感染的其他宿主蛋白的Interactome，我们发现Sirpa是一个细胞表面蛋白抑制巨噬细胞吞噬的肿瘤，死细胞和红细胞的吞噬作用，还会减少感染。重要的是，与TRIM2不同，SIRPA抑制了各种人的感染需要运输到酸性室的病毒病毒，包括VSV，Zika病毒， LCMV和埃博拉病毒和SARS-COV-2伪病毒。我们的数据表明TRIM2和SIRPA ACT 在病毒进入/内部化步骤中。这些发现表明有常见的机制调节病毒内吞作用和吞噬作用。我们在这里建议进一步研究病毒介导的内吞作用之间的重叠体外吞噬作用，体内和体内三个目标，这将1）研究重叠 NWA进入和吞噬作用途径； 2）确定trim2/sirpa抑制的位置感染发生； 3）使用trim2，sirpa和其他相关敲除小鼠探测角色这些蛋白质在细胞型特异性和体内感染中通过复制能力的NWA。在除了提供对NWAS进入细胞的机械洞察力的洞察力，这些研究还具有对宿主因素如何限制感染的潜力，并可能导致治疗干预的新方法。