Host factors and viral determinants mediating flavivirus NS1 tissue-specific endothelial dysfunction and vascular leak

介导黄病毒 NS1 组织特异性内皮功能障碍和血管渗漏的宿主因素和病毒决定因素

基本信息

批准号：
10417735
负责人：
Eva Harris
金额：
$ 68.63万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
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：
2022-04-18 至 2027-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10417735
关键词：
Address Affect Animal Model Binding Biochemical Genetics Biological Biological Markers Biological Response Modifiers Biology Blood Blood - brain barrier anatomy Blood Vessels Brazil Cathepsin L Cell Communication Cells Clathrin Clinical Complement Data Dengue Dengue Fever Deposition Disease Endocytosis Endothelial Cells Endothelium Enzymes Event Exhibits Family Fatal Outcome Flaviviridae Flavivirus Functional disorder Genetic Techniques Genomics Glycobiology Glycocalyx HIV Human In Vitro Individual Infection Integration Host Factors Intercellular Junctions Intervention Japanese Encephalitis Japanese encephalitis virus Lipoproteins Liver Mediating Medical Membrane Molecular Molecular Virology Mus Mutation Neuraminidase Nicaragua Nonstructural Protein Pathogenesis Pathogenicity Pathology Pathway interactions Patients Polysaccharides Process Proteins Public Health Role Sampling Serum Severity of illness Signal Pathway Signal Transduction Specificity Structural Biochemistry System Testing Tissues Tropism Variant Vietnam Viral Virus Virus Diseases Virus Replication West Nile virus Work Yellow Fever Yellow fever virus ZIKA Zika Virus brain endothelial cell burden of illness clinical investigation cytokine dimer endothelial dysfunction enzyme pathway heparanase human disease human pathogen improved in vivo mosquito-borne mouse model mutant neurotropic novel potential biomarker receptor severe dengue structural biology therapeutic target tissue tropism

项目摘要

Host factors and viral determinants mediating flavivirus NS1 tissue-specific endothelial dysfunction and vascular leak ABSTRACT The flavivirus (FV) genus contains medically important mosquito-borne human pathogens that cause a major global disease burden. While dengue (DENV), yellow fever (YFV), and Zika (ZIKV) viruses are systemic, and West Nile (WNV), Japanese encephalitis (JEV), and Zika viruses cause neurotropic infections, each FV can cause severe disease characterized in part by endothelial barrier dysfunction – the most classic example being vascular leak in severe dengue. This may result from overproduction of vasoactive cytokines as well as viral factors. The highly conserved FV non-structural protein 1 (NS1) is secreted from infected cells and circulates in the blood of infected humans. We and others have shown that FV NS1 can trigger endothelial barrier disruption in vitro and vascular leak in mice, independently from virus infection. In our current R01, we showed that endo- cytosis of FV NS1 into endothelial cells (ECs) followed by activation of key enzymes such as cathepsin L and heparanase leads to disruption of the endothelial glycocalyx layer (EGL) as well as mislocalization of intercellular junction proteins, both critical for maintaining endothelial barrier integrity. Interestingly, we found that FV NS1 proteins display exquisite tissue tropism, triggering EC dysfunction in vitro and in vivo in a manner reflecting tissue tropism and disease manifestations of each virus. While FV NS1 tissue tropism was determined by differential EC binding and internalization, downstream activation of key enzymes and signaling pathways required for pathogenesis appear to be conserved across FVs. However, host factors, viral determinants, and mechanisms mediating these processes are unknown. We hypothesize that distinct host factors on tissue- specific ECs mediate FV NS1 cell binding and internalization, leading to endothelial barrier dysfunction, virus dissemination, and different FV disease manifestations. In contrast, once a FV NS1 protein is internalized, we hypothesize that downstream steps of EC dysfunction are comparable – thus pointing the way to a pan-FV intervention. Here, we expand our previous work by identifying and characterizing host glycans, proteins, and NS1 determinants required for tissue-specific cell binding and internalization of NS1 in human ECs, mouse models, and clinical samples. We also define common mechanisms by which FV NS1 proteins trigger pathology. In Aim 1, we will identify and characterize host glycans and FV NS1 determinants required for differ- ential binding to tissue-specific ECs. In Aim 2, we will identify proteinaceous NS1 receptors required to initiate EC dysfunction and define mechanisms by which FV NS1 proteins mediate disruption of the EGL and intercellular junctions in tissue-specific ECs in vitro and in vivo. Aim 3 investigates the impact of FV NS1-mediated endothelial dysfunction on FV dissemination and pathogenesis in mouse models and human clinical samples from severe dengue and YF patients in Vietnam, Nicaragua and Brazil. This work is supported by experts in glycobiology, FV structural biology and biochemistry, vascular biology, FV pathogenesis and animal models, and clinical investigation and should identify biomarkers of severe FV disease and novel viral and host therapeutic targets.

宿主因素和病毒决定剂介导黄素NS1组织特异性内皮功能障碍和血管叶抽象的黄病毒（FV）属含有医学上重要的蚊子传播的人类病原体，引起主要的病原体全球疾病伯恩。风扇（DENV），黄热病（YFV）和Zika（Zikv）病毒是全身性的，并且西尼罗河（WNV），日本脑炎（JEV）和寨卡病毒引起神经感染，每个FV都可以导致部分由内皮屏障功能障碍部分特征的严重疾病 - 最经典的例子是严重风扇的血管泄漏。这可能是由于血管活性细胞因子和病毒的过量产生因素。高度组成的FV非结构蛋白1（NS1）是从感染细胞中分泌的，并在感染人类的血。我们和其他人表明，FV NS1可以触发内皮屏障破坏小鼠体外和血管泄漏，独立于病毒感染。在当前的R01中，我们表明 FV NS1的细胞增生到内皮细胞（EC），然后激活关键酶，例如组织蛋白酶L和肝素酶导致内皮糖脂层（EGL）的破坏以及细胞间的错误定位连接蛋白，既对维持内皮屏障完整性至关重要。有趣的是，我们发现FV NS1 蛋白质以反映的方式在体外和体内触发EC功能障碍表现出独特的组织向上主义每种病毒的组织向上主义和疾病表现。虽然FV NS1组织向逆转由差异EC结合和内在化，关键酶的下游激活和信号通路发病机理所要求的似乎是在FV上配置的。但是，宿主因素，病毒决定者和介导这些过程的机制尚不清楚。我们假设组织上的不同宿主因素 - 特定EC介导FV NS1细胞结合和内在化，导致内皮屏障功能障碍，病毒传播和不同的FV疾病表现。相反，一旦FV NS1蛋白被内化，我们假设EC功能障碍的下游步骤是可比的 - 从而指出了pan-fv的道路干涉。在这里，我们通过识别和表征宿主聚糖，蛋白质，扩展我们以前的工作 NS1决定了人类EC中NS1的组织特异性细胞结合和内在化所需的所需模型和临床样品。我们还定义了FV NS1蛋白触发的常见机制病理。在AIM 1中，我们将识别和表征宿主聚糖，而FV NS1确定了不同 - 与组织特异性EC的整个结合。在AIM 2中，我们将确定启动所需的蛋白质NS1受体 EC功能障碍和定义FV NS1蛋白介导EGL和细胞间破坏的机制组织特异性EC在体外和体内的连接。 AIM 3研究FV NS1介导的内皮的影响小鼠模型中FV传播和发病机理的功能障碍以及严重的人类临床样本越南，尼加拉瓜和巴西的登革热和YF患者。这项工作得到了糖生物学专家，FV的支持结构生物学和生物化学，血管生物学，FV发病机理和动物模型以及临床研究并应确定严重FV疾病以及新型病毒和宿主治疗靶标的生物标志物。