Understanding the Molecular Basis of Translation Inhibition by SARS-CoV-2 NSP14 and its Role in SARS-CoV-2 Immune Evasion

了解 SARS-CoV-2 NSP14 翻译抑制的分子基础及其在 SARS-CoV-2 免疫逃避中的作用

• 批准号: 10427688
• 负责人: Chun Chieh Hsu
• 金额: $ 15.75万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-22 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10427688
• 关键词: 2019-nCoV; Advisory Committees; Affect; Antigen Presentation; Antigen Presentation Pathway; Antiviral Agents; Antiviral Response; Bioinformatics; Biology; CD8-Positive T-Lymphocytes; COVID-19; COVID-19 pandemic; COVID-19 pathogenesis; COVID-19 therapeutics; Cell surface; Cellular Immunity; Cessation of life; Communicable Diseases; Data; Detection; Development; Enzymes; Etiology; Genes; Goals; Grant; Guanine; Guanosine; Head; Health; Histocompatibility Antigens Class I; Immune Evasion; Immune response; Immunobiology; Immunologic Surveillance; Immunology; In Vitro; Infection; Innate Immune Response; Interferon Type I; Interferons; Knowledge; Laboratory Research; Major Histocompatibility Complex; Manuscripts; Mediating; Messenger RNA; Methylation; Methyltransferase; Modification; Molecular; Nonstructural Protein; Play; Positioning Attribute; Protein Biosynthesis; Proteins; Publications; Publishing; RNA; Research; Research Personnel; Role; SARS-CoV-2 infection; SARS-CoV-2 inhibitor; SARS-CoV-2 pathogenesis; Surface; System; T cell response; T-Lymphocyte; Training; Translational Regulation; Translational Repression; Translations; Viral; Virus; Virus Diseases; antiviral drug development; antiviral immunity; blocking factor; career; cytotoxic CD8 T cells; design; effective therapy; global health; improved; insight; interest; interferon antagonist; mRNA Stability; mRNA Translation; novel vaccines; pathogenic virus; posttranscriptional; prevent; response; skills; therapeutic target; virology; virus host interaction

PROJECT SUMMARY/ABSTRACT The COVID-19 pandemic has had a devastating worldwide impact on health and economy. There is still no effective treatment, and the full extent of COVID-19 pathogenesis remains unclear. In particular, mechanisms for SARS-CoV-2 evasion of host immune surveillance remain poorly understood. I am interested in exploring both SARS-CoV-2 viral factors that block cellular translation and their roles in viral immune evasion. I have discovered that SARS-CoV-2 nonstructural protein 14 (NSP14) inhibits host translation and subsequently suppresses the innate immune response. Furthermore, my data suggest that the guanine-N7- methytransferase (N7-MTase) activity of NSP14 is required to inhibit translation, which catalyzes N7- methylguanosine (m7G) modification at 5’ cap guanosine. However, how m7G modification restricts cellular translation is unclear. Moreover, our data also showed that NSP14 inhibits the expression of MHC-I molecules on the cell surface and this also depends on its N7-MTase activity. However, whether NSP14 dampens MHC-I antigen presentation and cytotoxic CD8+ T cell responses requires further study. I hypothesize that NSP14 induces RNA m7G modification in SARS-CoV-2 infection and that shuts down cellular translation. I further hypothesize that such activity restricts the MHC-I antigen presentation pathway to escape T cell responses. The central objectives in this proposal are to define the molecular mechanism by which SARS-CoV-2 NSP14 inhibits cellular translation and enhance our understanding of how SARS-CoV-2 escapes T cell-mediated immunity. In Aim 1, I will determine the role of RNA m7G modification in translation inhibition in SARS-CoV-2 infection. In Aim 2, I will define how NSP14 restricts MHC-I antigen presentation and T cell responses. These two aims will address how SARS-CoV-2 infection can affect T cell-mediated immunity. I expect that our findings will uncover new strategies to develop new antiviral therapeutics for COVID-19 and help gain new insights into understanding the biology of COVID-19 pathogenesis. My career goal is to become an independent investigator studying virus-host interaction of infectious diseases, with a special focus on translational regulation and viral immune evasion. The proposed K22 grant will provide me with advanced training and skills to build specific expertise necessary to execute the proposed project and become independent in this field, including expertise in: 1. Immunology and Virology, 2. post- transcriptional modification and translational regulation research, and 3. skills necessary to head an independent research laboratory. To achieve these goals, I have assembled a Professional/Scientific Advisory Committee consisting of experts in SARS-CoV-2 Virology, Immunobiology, RNA Biology, and Bioinformatics.

项目总结/摘要 COVID-19疫情对全球健康及经济造成毁灭性影响。还有 没有有效的治疗方法，COVID-19发病机制的完整程度仍不清楚。特别是， SARS-CoV-2逃避宿主免疫监视的机制仍然知之甚少。我感兴趣 在探索SARS-CoV-2病毒因子阻断细胞翻译及其在病毒免疫中的作用方面， 逃避我发现SARS-CoV-2非结构蛋白14（NSP 14）抑制宿主翻译， 随后抑制先天免疫反应。此外，我的数据表明，鸟嘌呤-N7- NSP 14的甲基转移酶（N7-MTase）活性是抑制翻译所必需的，N7-MTase催化N7-甲基转移酶（N7-MTase）活性。 甲基鸟苷（m7 G）修饰。然而，m7 G修饰如何限制细胞 翻译不清楚。此外，我们的数据还表明，NSP 14抑制MHC-I分子的表达， 在细胞表面，这也取决于其N7-MTase活性。然而，NSP 14是否抑制MHC-I， 抗原呈递和细胞毒性CD 8 + T细胞应答需要进一步研究。我假设NSP 14 在SARS-CoV-2感染中诱导RNA m7 G修饰并关闭细胞翻译。我进一步 假设这种活性限制了MHC-I抗原呈递途径以逃避T细胞应答。 该提案的中心目标是确定SARS-CoV-2 NSP 14 抑制细胞翻译，并增强我们对SARS-CoV-2如何逃避T细胞介导的 免疫力在目的1中，我将确定RNA m7 G修饰在SARS-CoV-2翻译抑制中的作用 感染在目标2中，我将定义NSP 14如何限制MHC-I抗原呈递和T细胞应答。这些 两个目标将解决SARS-CoV-2感染如何影响T细胞介导的免疫。我希望我们的 研究结果将揭示新的策略，以开发新的抗病毒治疗COVID-19，并帮助获得新的 深入了解COVID-19发病机制的生物学。 我的职业目标是成为一名独立的研究者，研究感染性疾病的病毒-宿主相互作用。 疾病，特别关注翻译调节和病毒免疫逃避。K22计划 将为我提供高级培训和技能，以建立执行拟议的 项目，并成为独立在这一领域，包括专业知识：1。免疫学和病毒学，2。后 转录修饰和翻译调控研究; 3.领导一个 独立研究实验室。为了实现这些目标，我组建了一个专业/科学咨询小组， 委员会由SARS-CoV-2病毒学、免疫生物学、RNA生物学和生物信息学专家组成。