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

基本信息

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
项目类别：
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&apos Untranslated Regions 5&apos 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

项目摘要

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.

项目摘要/摘要在过去的二十年中，包括SARS-COV-2在内的三个高致病性人冠状病毒具有出现了，认为新的或重新出现的冠状病毒和其他RNA病毒的未来致命大流行是几乎不可避免。迫切需要治疗这些致命的传染病，将发现优先考虑开发新的有效抗病毒药。但是，尽管为解决Covid-19做出了前所未有的努力由于缺乏机械理解，危机，高效的Covid-19治疗学受到严重限制冠状病毒复制和发病机理。当前的大多数研究和治疗性开发工作专注于SARS-COV-2 RNA聚合酶（RDRP）或SPIKE蛋白，但它们的成功可能会严重破坏通过一种独特的SARS-COV-2校对机制，该机制掺入了RDRP抑制剂或快速耐药峰值蛋白突变体的出现。因此，必须开发创新的抗病毒针对病毒生命周期的不同的基本组成部分，针对不同的策略，在某些情况下尚未确定。复杂的病毒宿主相互作用构成了一个复杂的调节网络，该网络决定了病毒的结果感染，为创新的抗病毒疗法提供了有希望的机会。但是，当前对冠状病毒 - 宿主相互作用的知识主要仅限于激活或阻止的病毒宿主武器竞赛依赖干扰素的信号通路或竞争主机翻译机械。相比之下，病毒 - 冠状病毒基因表达和调节中的宿主相互作用在很大程度上是未知的领域。这个主要差距很大阻碍了针对这些备受掩盖的冠状病毒宿主相互作用的新型抗病毒剂的设计对于病毒生存至关重要，宿主抗病毒反应。该提议的主要目标是定义多个最近发现的SARS-COV-2-host相互作用的新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-host相互作用与病毒基因调控的新机制有关。结构生物学，蛋白-RNA的结合生物化学，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完成的任务。在AIM 1中，我们将确定分子和结构基础宿主因子与病毒复制转录机械之间的这些新发现的相互作用。在AIM 2中，我们将描述这些至关重要的宿主因素在调节病毒复制和转录时的生物学功能。在AIM 3中，我们将开发一个创新的捕获识别 - 可视化实验管道，以发现新的宿主因素和新型病毒复制转录调节机制。这些研究将在一起建立一个创新的概念框架来研究冠状病毒 - 宿主相互作用并揭示新目标为了开发新型抗癌病毒疗法。