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

• 批准号: 10362439
• 负责人: SUSAN R ROSS
• 金额: $ 44.11万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 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/10362439
• 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; Erythrocytes; Family; Functional disorder; Genes; Glycoproteins; Goals; Human; Human Pathology; Image; Immune response; Immune system; In Vitro; Incidence; Infection; Integration Host Factors; Junin virus; Knockout Mice; Lead; Link; Lymphocytic choriomeningitis virus; Mediating; Modeling; Mus; Mutation; Old World Arenaviruses; PTPN6 gene; PTPNS1 gene; Pathogenicity; Pathway interactions; Patients; Phagocytosis; 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; Time; Tissue Extracts; Ubiquitination; Vaccines; Vesicular stomatitis Indiana virus; Viral; Viral Hemorrhagic Fevers; Virus; Virus Diseases; Work; Zika Virus; Zoonoses; cell type; cellular targeting; cytokine; experimental study; in vivo; insight; knock-down; macrophage; member; mortality; neoplastic cell; nervous system disorder; novel; novel strategies; novel therapeutic intervention; pathogenic virus; prevent; receptor; therapeutic target; therapeutically effective; trafficking; tumor; ubiquitin ligase

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.

新世界出血热沙粒病毒 (NWA)，例如胡宁病毒，通过啮齿动物传播 当病毒传染给人类时，会导致约 30% 的死亡率。 NWA 的机制 诱发疾病仍不确定，尽管它可能包括诱导高水平的细胞因子 由免疫系统感染的前哨细胞产生，导致内皮细胞和血小板 功能障碍和神经系统疾病。胡宁感染的幸存者发展出强大的体液 免疫反应，表明在感染后早期控制感染对于 病毒清除。尽管有效的胡宁病毒疫苗降低了疾病发病率， 这种情况以及其他已知和新型 NWA 的零星病例没有疫苗可预防 或者仍然存在有效的治疗方法。众所周知，进化枝 B 致病性 NWA 结合 转铁蛋白受体 1 和细胞表面的其他受体，但导致它们进入的步骤 来自酸性细胞区室的作用尚未明确。我们最近进行了 siRNA 使用带有致病性 Junín 糖蛋白的假型病毒进行筛选，目的是发现 参与进入的宿主基因可以作为治疗靶点。我们发现TRIM2，一个 三方基序家族的成员，其中包括宿主内在的众所周知的成员 防御病毒感染，限制 NWA 内吞进入细胞。通过探测 TRIM2 对于阻止 NWA 感染的其他宿主蛋白的相互作用组，我们发现 SIRPA，一种细胞 抑制巨噬细胞吞噬肿瘤、死细胞和红细胞的表面蛋白， 也减少感染。重要的是，与 TRIM2 不同，SIRPA 可以抑制多种人类感染 需要运输到酸性区室的病原病毒，包括 VSV、寨卡病毒、 LCMV 和埃博拉病毒以及 SARS-Cov-2 假病毒。我们的数据表明 TRIM2 和 SIRPA 发挥作用 在病毒进入/内化步骤。这些发现表明存在共同的机制 调节病毒的内吞作用和吞噬作用。 我们在此建议进一步研究病毒介导的内吞作用之间的重叠 以及体外、离体和体内吞噬作用的三个目标，这将 1) 研究 NWA 进入和吞噬途径； 2) 确定 TRIM2/SIRPA 抑制的位置 发生感染； 3）使用TRIM2、SIRPA等相关基因敲除小鼠来探究其作用 这些蛋白质在细胞类型特异性和具有复制能力的 NWA 的体内感染中的作用。在 除了提供 NWA 进入细胞的机制见解外，这些研究还 增加我们对宿主因素如何限制感染并可能导致 治疗干预的新方法。