Emerging mechanisms of viral gene regulation from battles between host and SARS-CoV-2

宿主与 SARS-CoV-2 之间的战斗中病毒基因调控的新机制

• 批准号: 10725416
• 负责人: Chang Liu
• 金额: $ 46.89万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10725416
• 关键词: 2019-nCoV; 3' Untranslated Regions; 5' Untranslated Regions; Active Sites; Address; Affect; Affinity Chromatography; Antiviral Agents; Antiviral Response; Binding; Biological; Biological Models; Biological Process; Biology; Biophysics; COVID-19 pandemic; COVID-19 therapeutics; Cells; Color; Communicable Diseases; Complex; Coronavirus; Cryo-electron tomography; Cryoelectron Microscopy; DNA-Directed RNA Polymerase; Deposition; Development; Drug resistance; Economics; Future; GPRC5C gene; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Host Defense Mechanism; In Situ; Integration Host Factors; Interferons; Intervention; Kinetics; Knowledge; Life Cycle Stages; Link; Mass Spectrum Analysis; Mediating; Methyltransferase; Modification; Molecular; Outcome; Pathogenesis; Pathogenicity; Process; Proteins; RNA Biochemistry; RNA Polymerase Inhibitor; RNA Viruses; RNA chemical synthesis; RNA-Directed RNA Polymerase; Regulation; Replication-Associated Process; Replicon; Reporter; Research; Role; SARS-CoV-2 genome; Signal Pathway; Stream; Surface; System; Therapeutic; Time; Transcription Initiation; Transcription Process; Translations; Tretinoin; Vaccines; Viral; Viral Genes; Viral Pathogenesis; Virus; Virus Diseases; Virus Replication; Visualization; Work; arms race; biochemical tools; coronavirus therapeutics; design; experimental study; global health; human coronavirus; innovation; mutant; new therapeutic target; novel; novel coronavirus; novel strategies; novel therapeutics; pandemic disease; programs; reconstitution; reconstruction; research and development; sensor; single molecule; single-molecule FRET; structural biology; success; targeted agent; therapeutic development; therapeutically effective; tool; viral RNA; virology; virus host interaction

PROJECT SUMMARY/ABSTRACT Over the last two decades, three highly pathogenic human coronaviruses, including SARS-CoV-2, have emerged, arguing that future deadly pandemics of new or re-emerging coronaviruses and other RNA viruses are almost inevitable. The urgent need to treat these fatal infectious diseases has prioritized the discovery and development of novel effective antivirals. However, despite the unprecedented efforts to address the COVID-19 crisis, highly effective COVID-19 therapeutics are severely limited due to the lack of a mechanistic understanding of coronavirus replication and pathogenesis. Most of the current research and therapeutic development efforts focus on SARS-CoV-2 RNA polymerase (RdRp) or spike protein, but their success can be severely undermined by a unique SARS-CoV-2 proofreading mechanism that excises incorporated RdRp inhibitors or by the rapid emergence of drug-resistant spike protein mutants. Therefore, it is imperative to develop innovative antiviral strategies targeting distinct, and in some cases yet to be identified, essential components of the viral life cycle. The intricate virus-host interplay constitutes a sophisticated regulatory network that dictates the outcome of virus infection, affording promising opportunities to be explored for innovative antiviral therapeutics. However, current knowledge of coronavirus-host interactions is mostly limited to the virus-host arms race in activating or blocking the interferon-dependent signaling pathways or in competing for host translation machinery. By contrast, virus- host interplay in coronavirus gene expression and regulation is largely uncharted territory. This major gap greatly hinders the design of novel antiviral agents targeting these much-overlooked coronavirus-host interactions that are critical for viral survival and host antiviral responses. The central objectives of this proposal are to define novel molecular mechanisms and functional roles of multiple recently discovered SARS-CoV-2-host interactions in modulating the viral replication and transcription processes and to identify new SARS-CoV-2-host interactions linked to novel mechanisms of viral gene regulation. With a combination of structural biology, protein-RNA biochemistry, single-molecule biophysics, cell virology, and computational approaches, we will accomplish these tasks through three tightly interwoven aims. In Aim 1, we will determine the molecular and structural basis of these newfound interactions between host factors and the viral replication-transcription machinery. In Aim 2, we will delineate the biological functions of these crucial host factors in modulating viral replication and transcription. In Aim 3, we will develop an innovative capture-identification-visualization experimental pipeline to discover new host factors and novel viral replication-transcription-regulating mechanisms. Together, these studies will establish an innovative conceptual framework to study coronavirus-host interactions and reveal new targets for the development of novel anti-coronavirus therapeutics.

项目总结/摘要 在过去的二十年里，包括SARS-CoV-2在内的三种高致病性人类冠状病毒， 他们认为，未来新的或重新出现的冠状病毒和其他RNA病毒的致命大流行， 几乎是不可避免的治疗这些致命传染病的迫切需要优先考虑发现和 开发新的有效的抗病毒药物。然而，尽管为应对COVID-19做出了前所未有的努力， 危机，由于缺乏对机制的理解，高效的COVID-19治疗方法受到严重限制 冠状病毒复制和发病机理的研究。目前的大多数研究和治疗开发工作 目前，研究重点是SARS-CoV-2 RNA聚合酶（RdRp）或刺突蛋白，但它们的成功可能会受到严重破坏 通过一种独特的SARS-CoV-2校对机制，切除掺入的RdRp抑制剂，或通过快速的 耐药刺突蛋白突变体的出现。因此，开发创新的抗病毒药物势在必行 针对病毒生命周期的不同（在某些情况下尚待确定）基本组成部分的战略。 复杂的病毒-宿主相互作用构成了一个复杂的调控网络，决定了病毒的结果。 感染，提供有希望的机会，以探索创新的抗病毒治疗。但目前的 关于冠状病毒与宿主相互作用的知识大多局限于病毒与宿主在激活或阻断 干扰素依赖性信号通路或竞争宿主翻译机制。相反，病毒- 宿主在冠状病毒基因表达和调控中的相互作用在很大程度上是未知领域。这一重大差距大大 阻碍了针对这些被忽视的冠状病毒与宿主相互作用的新型抗病毒药物的设计， 对病毒存活和宿主抗病毒反应至关重要。本建议的主要目标是确定 新发现的SARS-CoV-2与宿主相互作用的分子机制和功能作用 在调节病毒复制和转录过程中，并确定新的SARS-CoV-2-宿主相互作用 与病毒基因调控的新机制有关。结合结构生物学，蛋白质-RNA 生物化学，单分子生物物理学，细胞病毒学和计算方法，我们将完成这些 通过三个紧密交织的目标。在目标1中，我们将确定 这些新发现的宿主因子和病毒复制-转录机制之间的相互作用。在目标2中， 将描述这些重要的宿主因子在调节病毒复制和转录中的生物学功能。 在目标3中，我们将开发创新的捕获-识别-可视化实验管道，以发现新的 宿主因子和新的病毒复制-转录-调节机制。这些研究将 建立创新的概念框架，研究冠状病毒与宿主的相互作用，并揭示新的靶点 用于开发新型抗冠状病毒疗法。