Molecular Mechanisms of Filoviral-host Interactions

丝病毒-宿主相互作用的分子机制

• 批准号: 10555051
• 负责人: Gaya K. Amarasinghe
• 金额: $ 335.5万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-07 至 2028-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10555051
• 关键词: Academia; Actins; Address; Antibodies; Antibody Therapy; Biochemical; Biochemistry; Biological; Biological Assay; Biology; Cell Culture Techniques; Cell Line; Cells; Cellular biology; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Complex; Computer Assisted; Containment; Cryoelectron Microscopy; Data; Development; Disease; Disease Outbreaks; Distant; Ebola virus; Emerging Communicable Diseases; Ensure; Evaluation; Family; Family member; Filoviridae Infections; Filovirus; Funding; Genetic; Genetic Transcription; Goals; Growth; Immune response; Immunology; Industry; Infection; Integration Host Factors; Knockout Mice; Knowledge; Laboratories; Learning; Maps; Marburg Virus Disease; Marburgvirus; Mass Spectrum Analysis; Methods; Modeling; Molecular; Mutation; Nucleocapsid; Pathogenesis; Pathway Analysis; Pathway interactions; Periodicals; Phenotype; Physical condensation; Play; Predisposition; Productivity; Program Research Project Grants; Progress Reports; Protein-Protein Interaction Map; Proteins; Proteomics; Public Health; Publications; RNA Processing; Reagent; Records; Research; Research Personnel; Research Project Grants; Resource Sharing; Resources; Rodent; Role; Scientist; Set protein; System; Testing; Therapeutic; Therapeutic Intervention; Transgenic Organisms; Vaccines; Viral; Viral Proteins; Virion; Virus; Virus Assembly; Virus Diseases; Virus Replication; Virus-like particle; Work; Zoonoses; cell type; conditional knockout; data complexity; data sharing; emerging pathogen; experience; human disease; in vivo; innovation; insight; interdisciplinary approach; medical countermeasure; mouse model; new therapeutic target; novel; novel therapeutic intervention; pathogen; protein protein interaction; recruit; structural biology; targeted treatment; tomography; training opportunity; virology

Overall Project Summary/Abstract: Molecular mechanisms of filoviral-host interactions The family Filoviridae, which includes Ebola virus (EBOV) and Marburg virus (MARV), are zoonotic pathogens that cause outbreaks of severe human disease and require biosafety level 4 (BSL4) containment for study. Recent approval of a vaccine and antibody-based therapies against an EBOV represent progress towards medical countermeasures. However, the family is comprised of multiple antigenically distinct species, making identification of pan-filoviral therapeutic approaches desirable. Furthermore, the molecular mechanisms required for replication and pathogenesis are incompletely understood. Defining key filovirus-host interactions and the mechanisms by which they promote viral growth and disease will provide important insight into viral biology and suggest new therapeutic approaches. Existing data, including our own, have identified key host-viral interactions that likely play important roles in the pathogenesis of filovirus disease. Our overarching goal is to address this gap in knowledge by building and expanding upon the strong foundational knowledge on EBOV to define molecular mechanisms at the host-pathogen interface and to identify EBOV-specific and pan-filoviral interactions that contribute to pathogenesis. To achieve our goals, we have assembled a highly accomplished team with track records of effective synergistic collaboration and expertise ranging from molecular biochemistry, structural biology and mass spectrometry to cell biology, virology, and work at BSL4. In the current funding period, we identified multiple host pathways that impact EBOV infection and defined key interactions at the viral-host interface. In our proposed studies, we use a reductionist approach to define molecular mechanisms by biochemical and structural methods (Project 1; RP01), determine the cellular impact and contributions of viral proteins such as VP30 and VP24 in immune response, viral replication, assembly and egress (Project 2; RP02), and evaluate the impact of specific interactions with EBOV and MARV virus in cell culture and in vivo, including specific subnetworks that regulate filoviral entry and replication (Project 3; RP03). Recognizing the complexity of the data being generated we have recruited new expertise in proteomics and genetic network analysis to provide a deeper understanding of host-virus protein connectivity and interaction. These efforts will be further supported by two scientific cores, the Antibody and Reagent Development Core B and the BSL4/ABSL4 laboratory Core C. This work will be guided by an active Administrative Core A that will receive critical input from the Core A Advisory Group (CAAG) and the External Advisory Board (EAB). Each is comprised of preeminent scientists in academia and industry with strong productivity in emerging infectious diseases and immunology and significant advisory experience. Building on our productive initial work, we are poised to define a comprehensive host interaction network, validate regulatory mechanisms that drive viral infection, and identify targets for therapeutic intervention. Our unique innovative experimental framework and highly interactive scientific approach provides a blueprint to tackle other emerging and reemerging pathogens.

总体项目总结/摘要：丝状病毒-宿主相互作用的分子机制 丝状病毒科（Filoviridae）包括埃博拉病毒（Ebola virus，EBOV）和马尔堡病毒（Marburg virus，MARV 导致严重人类疾病爆发，需要生物安全4级（BSL 4）控制进行研究。 最近批准的针对EBOV的疫苗和基于抗体的疗法代表了朝着 医学对策然而，该家族由多个抗原性不同的物种组成， 鉴定所需的泛丝状病毒治疗方法。此外，所需的分子机制 对复制和发病机制的了解还不完全。定义关键的丝状病毒-宿主相互作用和 它们促进病毒生长和疾病的机制将为病毒生物学和 提出了新的治疗方法。现有的数据，包括我们自己的数据，已经确定了关键的宿主-病毒相互作用 可能在丝状病毒病的发病机制中起重要作用。我们的首要目标是解决这个问题 通过建立和扩展关于EBOV的强大基础知识， 宿主-病原体界面的分子机制，并确定EBOV特异性和泛丝状病毒相互作用 导致发病机制。为了实现我们的目标，我们组建了一个非常成功的团队， 有效的协同合作和专业知识的跟踪记录，从分子生物化学，结构， 生物学和质谱到细胞生物学，病毒学，并在BSL 4工作。在目前的融资期内，我们 确定了影响EBOV感染的多种宿主途径，并确定了病毒-宿主之间的关键相互作用 接口.在我们提出的研究中，我们使用还原论的方法来定义分子机制， 生物化学和结构方法（项目1; RP 01），确定细胞的影响和贡献的病毒 免疫应答、病毒复制、装配和排出中的蛋白质如VP 30和VP 24（项目2; RP 02）， 并评估与EBOV和MARV病毒在细胞培养和体内的特异性相互作用的影响，包括 调节丝状病毒进入和复制的特定亚网络（项目3; RP 03）。认识到 我们已经招募了蛋白质组学和遗传网络分析方面的新专业知识， 提供宿主病毒蛋白质连接和相互作用的更深入的了解。这些努力将进一步 由两个科学核心支持，抗体和试剂开发核心B和BSL 4/ABSL 4 实验室核心C。这项工作将由一个积极的行政核心A指导，该核心A将接受以下方面的重要投入： 核心A咨询小组（CAAG）和外部咨询委员会（EAB）。每一个都由杰出的 学术界和工业界的科学家在新兴传染病和免疫学方面具有强大的生产力， 重要的咨询经验。在我们富有成效的初步工作的基础上，我们准备确定一个全面的 宿主相互作用网络，验证驱动病毒感染的调节机制，并确定 治疗干预我们独特的创新实验框架和高度互动的科学 该方法为应对其他新出现和重新出现的病原体提供了蓝图。

项目成果

Filovirus Strategies to Escape Antiviral Responses.

• DOI: 10.1007/82_2017_13
• 发表时间: 2017
• 期刊: Current topics in microbiology and immunology
• 作者: Olejnik J;Hume AJ;Leung DW;Amarasinghe GK;Basler CF;Mühlberger E
• 通讯作者: Mühlberger E

Human IFIT3 Modulates IFIT1 RNA Binding Specificity and Protein Stability.

• DOI: 10.1016/j.immuni.2018.01.014
• 发表时间: 2018-03-20
• 期刊: Immunity
• 影响因子: 32.4
• 作者: Johnson B;VanBlargan LA;Xu W;White JP;Shan C;Shi PY;Zhang R;Adhikari J;Gross ML;Leung DW;Diamond MS;Amarasinghe GK
• 通讯作者: Amarasinghe GK

Ebola Virus Replication Stands Out.

埃博拉病毒复制脱颖而出。

• DOI: 10.1016/j.tim.2019.05.004
• 发表时间: 2019
• 期刊: Trends in microbiology
• 影响因子: 15.9
• 作者: Wang,Wenjie;Wu,Chao;Amarasinghe,GayaK;Leung,DaisyW
• 通讯作者: Leung,DaisyW

A Novel Proximity Biotinylation Assay Based on the Self-Associating Split GFP1-10/11.

一种基于自缔约的分裂GFP1-10/11的新型接近生物素化测定法。

• DOI: 10.3390/proteomes8040037
• 发表时间: 2020-12-02
• 期刊: Proteomes
• 影响因子: 3.3
• 作者: Kesari AS;Aryal UK;LaCount DJ
• 通讯作者: LaCount DJ

When your cap matters: structural insights into self vs non-self recognition of 5' RNA by immunomodulatory host proteins.

• DOI: 10.1016/j.sbi.2016.02.001
• 发表时间: 2016-02
• 期刊: Current opinion in structural biology
• 影响因子: 6.8
• 作者: Leung DW;Amarasinghe GK
• 通讯作者: Amarasinghe GK