SARS-CoV-2 and Autophagy

SARS-CoV-2 和自噬

• 批准号: 10174059
• 负责人: William T Jackson
• 金额: $ 42.49万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-14 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10174059
• 关键词: 2019-nCoV; Autophagocytosis; Autophagosome; Biological Assay; COVID-19; COVID-19 pandemic; Cells; Cellular Membrane; Cellular biology; Chloroquine; Common Cold; Complement; Complex; Coronavirus; Coronavirus Infections; Data; Disease; Disease Outbreaks; Endosomes; Environment; Estrogen receptor positive; Family; Family Picornaviridae; Generations; Knowledge; Life Cycle Stages; Lipids; Lysosomes; Membrane; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Monitor; Nonstructural Protein; Paper; Pathway interactions; Play; Proteins; RNA replication; Respiratory distress; Role; SNAP receptor; Severe Acute Respiratory Syndrome; Site; Small Interfering RNA; Testing; Vesicle; Viral Genome; Virus; Virus Replication; Work; experience; expression cloning; human coronavirus; inhibitor/antagonist; member; new therapeutic target; paralogous gene; prevent; public health priorities; recruit; targeted treatment; tool; viral RNA; viral genomics; virus host interaction

PROJECT SUMMARY/ABSTRACT Coronaviruses are major causes of disease worldwide, from the common cold to outbreaks of strains causing severe and sometimes lethal respiratory distress, including SARS, MERS, and the current outbreak of COVID- 19 caused by SARS-CoV-2. It has been observed over the last two decades that multiple coronaviruses interact with components of the autophagy pathway to rearrange cellular membranes and generate sites for RNA replication. In this application, we propose to test the interactions of SARS-CoV-2 with the autophagy pathway. We will rapidly assess whether SARS-CoV-2 has similar interactions with the autophagy pathway as other coronaviruses, then extend the field by identifying how these interactions interfere with viral replication, providing novel therapeutic targets. We will carry this strategy out through three Aims: In the first, we will identify requirements from the early autophagy pathway for SARS-CoV-2 replication. Data from coronavirus studies suggest that the autophagy protein LC3 is recruited to generate double membraned vesicles for virus genomic RNA replication. Other data suggest LC3 is utilized through the ER-associated degradation (ERAD) machinery to generate replication vesicles. We will determine which, if either, of these two pathways is used by SARS-CoV-2. In the second aim, we will determine the role of vesicle acidification in SARS-CoV-2 replication. There is strong evidence that acidification of vesicles plays a role in multiple steps of CoV replication, and we will identify the role of vesicle acidification in SARS-CoV-2 replication. In the third aim, we will identify the specific roles of SARS-CoV-2 non-structural proteins in induction of the autophagic machinery. We have identified previous studies of expression of non-structural proteins in coronavirus and will use this to guide our study of SARS-CoV-2 proteins. These Aims complement existing studies of autophagy in RNA viral life cycles by the Jackson Lab while leveraging the existing expertise of the Frieman Lab in studying cell biology of coronavirus infections. By completing these Aims, we will understand whether SARS-CoV-2 interacts with the autophagy pathway in a similar or unique fashion compared to other coronaviruses, and extend the coronavirus field by identifying specific host-virus interactions that may be targeted for COVID-19 therapies.

项目摘要/摘要 冠状病毒是世界范围内疾病的主要原因，从普通感冒到引起 严重的、有时是致命的呼吸窘迫，包括SARS、MERS和目前爆发的COVID- 19例由SARS-CoV-2引起。在过去的二十年里，人们观察到多种冠状病毒 与自噬途径的组件相互作用，重新排列细胞膜并为其生成位点 RNA复制。在这个应用中，我们建议测试SARS-CoV-2与自噬的相互作用 路径。我们将迅速评估SARS-CoV-2是否与自噬途径具有类似的相互作用 其他冠状病毒，然后通过识别这些相互作用如何干扰病毒复制来扩展领域， 提供新的治疗靶点。我们将通过三个目标来实施这一战略：第一，我们将 确定SARS-CoV-2复制的早期自噬途径的要求。冠状病毒数据 研究表明，自噬蛋白lc3被招募来为病毒产生双膜泡。 基因组RNA复制。其他数据表明，通过内质网相关降解(ERAD)利用了LC3 产生复制小泡的机械。我们将确定这两条路径中的哪条路径由 SARS-CoV-2。在第二个目标中，我们将确定囊泡酸化在SARS-CoV-2复制中的作用。 有强有力的证据表明，囊泡的酸化在冠状病毒复制的多个步骤中发挥了作用，我们 将确定囊泡酸化在SARS-CoV-2复制中的作用。在第三个目标中，我们将确定 SARS-CoV-2非结构蛋白在自噬机制诱导中的特殊作用。我们有 确认了以前在冠状病毒中表达非结构蛋白的研究，并将利用这一点来指导我们的 SARS-CoV-2蛋白的研究这些目的是对现有关于RNA病毒生命周期中自噬的研究的补充 杰克逊实验室在利用弗里曼实验室现有的专业知识研究细胞生物学的同时 冠状病毒感染。通过完成这些目标，我们将了解SARS-CoV-2是否与 与其他冠状病毒相比，自噬途径类似或独特，并扩展了 通过识别可能成为新冠肺炎疗法目标的特定主机-病毒相互作用，我们可以在冠状病毒领域开展研究。