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.

项目摘要/摘要 COVID-19大流行对全球范围内对健康和经济产生了破坏性的影响。还有 没有有效的治疗方法，共同199的发病机理的全部范围尚不清楚。尤其， SARS-COV-2宿主免疫监视的演变的机制仍然很少理解。我很感兴趣 在探索两个SARS-COV-2病毒因子时，可以阻止细胞翻译及其在病毒免疫中的作用 逃避。我发现SARS-COV-2非结构蛋白14（NSP14）抑制宿主翻译和 随后抑制了先天的免疫反应。此外，我的数据表明鸟嘌呤-N7- 需要NSP14的甲基转移酶（N7-mTase）活性才能抑制翻译，从而催化N7-- 5'帽鸟嘌呤的甲基鸟苷（M7G）修饰。但是，M7G修改如何限制细胞 翻译不清楚。此外，我们的数据还表明，NSP14抑制MHC-I分子的表达 在细胞表面上，这也取决于其N7-mTase活性。但是，NSP14是否抑制MHC-I 抗原表现和细胞毒性CD8+ T细胞反应需要进一步研究。我假设NSP14 在SARS-COV-2感染中诱导RNA M7G修饰，并关闭细胞翻译。我进一步 假设这种活动限制了MHC-I抗原呈现途径以逃避T细胞反应。 该提案中的中心对象是定义SARS-COV-2 NSP14的分子机制 抑制细胞翻译并增强我们对SARS-COV-2如何逃脱T细胞介导的理解 免疫。在AIM 1中，我将确定RNA M7G修饰在SARS-COV-2中翻译抑制中的作用 在AIM 2中，我将定义NSP14如何限制MHC-I抗原表现和T细胞反应。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 调查结果将发现新的策略，以开发新的抗病毒疗法为Covid-19，并帮助获得新的策略 了解Covid-19发病机理的生物学的见解。 我的职业目标是成为研究传染病病毒宿主相互作用的独立研究者 疾病，特别关注翻译调节和病毒免疫驱虫。拟议的K22赠款 将为我提供高级培训和技能，以建立执行拟议的特定专业知识 项目并在该领域独立，包括：1。免疫学和病毒学的专业知识，2。 转录修改和翻译法规研究以及3。领导的技能 独立研究实验室。为了实现这些目标，我组装了专业/科学咨询 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的委员会。