Modulation of the cGAS/STING innate immune sensing pathway by arboviruses of human health interest

影响人类健康的虫媒病毒对 cGAS/STING 先天免疫传感通路的调节

• 批准号: 10012752
• 负责人: Lawrence Gabe Webb
• 金额: $ 4.39万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10012752
• 关键词: Aedes; Alphavirus; Americas; Antiviral Agents; Arboviruses; Area; Arthralgia; Arthropod Vectors; BCAR3 gene; Cells; Chikungunya virus; Chronic; Clinical; Complex; Computer Models; Culicidae; Cyclic GMP; DNA Polymerase II; Dendritic Cells; Dengue Infection; Dengue Vaccine; Dengue Virus; Detection; Development; Disease Outbreaks; Embryo; Encephalitis; Epidemic; Fever; Fibroblasts; Flavivirus; Future; Generations; Genes; Guillain-Barré Syndrome; Health; Hemorrhage; Hepatitis; Human; Immune; Incidence; Infection; Influenza; Innate Immune Response; Innate Immune System; Interferon Type I; Interferons; Laboratories; Link; Mediating; Meningoencephalitis; Methods; Microcephaly; Microscopy; Mitochondria; Molecular; Mus; Mutagenesis; Nonstructural Protein; Pathway interactions; Pattern; Peptide Hydrolases; Phosphorylation; Population; Primary Infection; Property; Proteins; Public Health; Publications; RNA; RNA Viruses; Reporter; Reporting; Research; Rest; Rice; Role; SH2D3C gene; STAT1 gene; STING agonists; Serotyping; Signal Transduction; Signaling Protein; Site; Small Interfering RNA; Stimulator of Interferon Genes; Study models; Symptoms; System; Temperature; Testing; Tretinoin; Universities; Vaccine Design; Vaccines; Viral; Viral Hemorrhagic Fevers; Viremia; Virus; Virus Diseases; Virus Replication; West Nile viral infection; West Nile virus; Work; Yellow fever virus; Zika Virus; base; chikungunya infection; falls; human pathogen; innate immune mechanisms; innate immune sensing; interest; knock-down; lens; member; monocyte; mosquito-borne; mouse model; novel; novel vaccines; overexpression; pathogen; protein protein interaction; prototype; response; screening; transmission process; vaccine development; vector; vector mosquito; viral detection

Project Summary Nearly half of the world’s population lives in areas where Aedes spp. mosquitos circulate, which can transmit several viruses of human health concern including dengue virus (DENV), Zika virus (ZIKV), yellow fever virus (YFV), West Nile virus (WNV), and chikungunya virus (CHIKV). There are no specific antivirals licensed for the treatment of these human pathogens and while the YFV vaccine provides hope for new vaccine development, there is still a need for the development of new or better vaccines for the rest of these viruses. These concerns came to the forefront during the 2015-2016 unprecedented epidemic of ZIKV in the Americas, when this virus was linked to cases of microcephaly and Guillain-Barré syndrome. For all viruses to replicate and spread, they must evade detection by the host’s innate immune system, in particular the type I interferon (type-I IFN) response. Previously, the Fernandez-Sesma laboratory has shown that the cGAS/STING innate immune sensing pathway can serve as a strong restriction factor for DENV, serotype-2, replication and that the NS2B3 protease complex and its components can antagonize both cGAS and STING. Additionally, work from the Rice laboratory at Rockefeller University has implicated that cGAS is an important restriction factor in controlling arbovirus replication. Furthermore, recent publications have illustrated that direct agonists of STING are able to potently restrict CHIKV replication. In this study, it is hypothesized that the cGAS/STING innate immune sensing pathway serves to restrict the replication of the specific human arboviral pathogens listed above and that these viruses have mechanisms of cGAS/STING antagonism. The primary objectives of this study are to determine the role of the cGAS/STING innate immune sensing pathway in antagonizing the replication of DENV, ZIKV, WNV, YFV, and CHIKV, and to identify novel viral antagonists of both cGAS and STING. By exogenous overexpression of viral components, Immunofluorescent microscopy, infection of primary cell systems, and type-I IFN reporter systems as well as computational modeling and siRNA knockdowns of the cGAS/STING and complementary pathways, this study will identify previously undiscovered arboviral antagonism of the cGAS/STING pathway, for the aforementioned arboviruses, and the restriction that both cGAS and STING impose upon the replication of these viruses. Information generated from this work has broad implications further challenging the dogma of investigating ssRNA viral infection through the lens of RNA pathogen associated molecular patterns (PAMPs) alone and expands that to include the study of viral antagonism of danger associated molecular patterns (DAMPs) induced during viral infection. Furthermore, this work can serve to inform the screening of virus specific antiviral compounds and aid in the design of vaccine prototypes.

项目摘要 世界上近一半的人口生活在伊蚊属的地区。蚊子四处传播， 几种人类健康问题病毒，包括登革热病毒（DENV）、寨卡病毒（ZIKV）、黄热病病毒 (YFV)、西尼罗河病毒（WNV）和基孔肯雅病毒（CHIKV）。没有特定的抗病毒药物许可用于 这些人类病原体的治疗，虽然YFV疫苗为新疫苗的开发提供了希望， 仍然需要开发针对其余这些病毒的新的或更好的疫苗。这些关切 在2015-2016年美洲前所未有的ZIKV流行期间， 与小头畸形和格林-巴利综合征病例有关。对于所有的病毒复制和传播，他们 必须逃避宿主先天免疫系统的检测，特别是I型干扰素（I型IFN） 反应此前，Fernando-Sesma实验室已经证明，cGAS/STING先天免疫 NS 2B 3是DENV血清型2复制的强限制因子， 蛋白酶复合物及其组分可以拮抗cGAS和STING两者。此外，Rice的工作 洛克菲勒大学的一个实验室暗示，cGAS是一个重要的限制因素， 虫媒病毒复制此外，最近的出版物已经说明STING的直接激动剂能够 有效地限制CHIKV复制。在这项研究中，假设cGAS/STING先天免疫 传感途径用于限制上述特定人虫媒病毒病原体的复制， 这些病毒具有cGAS/STING拮抗机制。本研究的主要目的是 确定cGAS/STING先天性免疫传感途径在拮抗肿瘤细胞复制中的作用。 DENV、ZIKV、WNV、YFV和CHIKV，并鉴定cGAS和STING两者的新型病毒拮抗剂。通过 外源性过表达病毒成分，免疫荧光显微镜，原代细胞感染 系统，和I型IFN报告系统，以及计算建模和siRNA敲低的干扰素， cGAS/STING和互补途径，这项研究将确定以前未发现的虫媒病毒 cGAS/STING途径的拮抗作用，以及对上述虫媒病毒的限制， cGAS和STING对这些病毒的复制施加影响。这项工作产生的信息 广泛的影响进一步挑战了通过RNA的透镜研究ssRNA病毒感染的教条 病原体相关分子模式（PAMPs），并将其扩展到包括病毒的研究 拮抗病毒感染期间诱导的危险相关分子模式（DAMP）。而且这 这项工作可以为筛选病毒特异性抗病毒化合物提供信息，并有助于设计疫苗。 原型