The Untranslated 3'End of SARS-CoV-2 RNA as a Determinant of Obesity-Accelerated Infectivity

SARS-CoV-2 RNA 的非翻译 3 末端是肥胖加速感染的决定因素

基本信息

批准号：
10689137
负责人：
PAUL L FOX
金额：
$ 40.25万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-14 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10689137
关键词：
2019-nCoV 3&apos Untranslated Regions ACE2 Acceleration Acute Adipocytes Adipose tissue Adult Affect Affinity Antisense RNA Antiviral Agents Binding Binding Proteins Biological Assay COVID-19 COVID-19 pandemic COVID-19 therapeutics Cell Surface Receptors Code Complex Effectiveness Elements Epidemic Epitopes Event Foundations Genome Genomics Hepatitis B Vaccines Hormones Human Hypersensitivity Immunocompromised Host Incidence Inflammatory Influenza B Virus Insulin Interferon Type II Knowledge Luciferases Messenger RNA Molecular Mutate Mutation Obesity Oligonucleotides Pathologic Pathway interactions Patients Polyproteins Population Proteins Proteomics RNA RNA-Binding Proteins RNA-Directed RNA Polymerase Regulation Regulatory Pathway Reporter Resistance Ribosomal Frameshifting Risk Factors Role SARS coronavirus SARS-CoV-2 inhibitor Series Stimulus Structural Protein Structure Syndrome Testing Therapeutic Therapeutic Agents Therapeutic Intervention Translational Repression Translations Untranslated RNA Untranslated Regions Vaccination Vaccines Viral Viral Genome Viral Nonstructural Proteins Viral Proteins Viral reservoir Virus Virus Assembly Virus Replication betacoronavirus cytokine demographics experimental study genomic RNA glutamyl-prolyl-tRNA synthetase improved inhibitor mRNA Translation mimetics new therapeutic target novel obese patients pandemic disease particle protein expression receptor targeted agent therapeutic RNA therapeutic development therapeutic target viral RNA

项目摘要

Project Summary/Abstract Obesity is an epidemic-scale problem in the U.S. affecting about 35% of the adult population, and is a major risk factor for the ongoing pandemic, COVID-19. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the RNA betacoronavirus that is the causative agent of COVID-19. Despite rapid progress in developing effective vaccines, progress in devising improved therapeutics has been slow, and there is an urgent need for anti-viral therapeutics for treatment of infected patients not protected by vaccines. Certain demographics are variably resistant to vaccination, for example, obesity markedly reduces effectiveness of several vaccines. Therapeutics can also be critical in the eventuality that mutations in the virus render the vaccine ineffective. The molecular events responsible for expression of SARS-CoV-2 proteins are known from studies of other betacoronaviruses; however, the regulatory pathways and pathological conditions determining their expression are poorly understood. The translation of viral RNA utilizes the host mRNA translation machinery, primarily regulated by specific RNA-binding proteins or complexes that bind sequence or structural elements in terminal non-coding regions. Importantly, the coding regions of SARS-CoV-2 genomic RNA and the ten subgenomic mRNAs (sgmRNAs) are likewise bordered by non-coding upstream and downstream regions, termed the 5'- leader and 3'-end sequences, respectively. A critical feature of the genome and the sgmRNAs of SARS-CoV-2 is that identical 5'-leader and 3'-end sequences are present in all. Thus, the terminal sequences represent novel, unexplored targets for interference with virus assembly and function. We have discovered a 39-nt sequence in the 3'-end of SARS-CoV-2 bearing structural similarity to the GAIT (interferon-gamma-activated inhibitor of translation) RNA element previously described by us. We show that glutamyl-prolyl tRNA synthetase (EPRS) a protein that binds the human GAIT element also binds the vGLE. Moreover, IFN-γ, a pro-inflammatory cytokine, and insulin, an obesity-induced hormone, markedly increases expression of a luciferase reporter bearing the intact vGLE. These results are the first to show a functional consequence of an RNA element in the 3'-end sequence of SARS-CoV-2. We hypothesize that binding of EPRS to the vGLE stimulates sgmRNA translation required for expression of structural and other SARS-CoV-2 proteins, and for programmed ribosomal frameshifting required for genome replication. We will test this hypothesis by pursuing two Specific Aims: In Aim 1 we elucidate the role of EPRS binding to the SARS-CoV-2 3'UTR vGLE in regulating viral replication and sgmRNA translation. In Aim 2 we develop RNA inhibitors that target EPRS/vGLE interactions and block viral protein expression. We anticipate these fundamental studies will provide the first information on the function of the 3'-end of SARS-CoV-2, and will provide a foundation for development of therapeutic agents to be used in combination with mechanistically distinct anti-viral agents targeting the vGLE and possibly emerging betacoronaviruses.

项目摘要/摘要在美国，肥胖是一个流行病的问题，影响了大约35％的成年人口，这是一个主要风险正在进行的大流行的因素，共同19。严重的急性呼吸综合症冠状病毒2（SARS-COV-2）为 RNA betacoronavirus是Covid-19的致病药物。尽管发展迅速有效的疫苗，改善治疗方面的进展一直很慢，迫切需要用于治疗未受疫苗保护的感染患者的抗病毒疗法。某些人口统计是例如，对疫苗的可变耐药性明显降低了几种疫苗的有效性。如果病毒的突变使疫苗无效，则治疗剂也可能至关重要。从其他的研究中知道，负责SARS-COV-2蛋白表达表达的分子事件 betacoronaviruses;但是，调节途径和病理状况决定其表达知之甚少。病毒RNA的翻译利用宿主mRNA翻译机制，主要由特定的RNA结合蛋白或结合终端中结合序列或结构元件的复合物调节非编码区域。重要的是，SARS-COV-2基因组RNA和十个亚基因组的编码区域 MRNA（SGMRNA）同样被非编码上游和下游区域所接加，称为5'-- 领导者和3'末端序列。 SARS-COV-2的基因组和SGMRNA的关键特征共同存在相同的5'-Leader和3'-End序列。那，终端序列代表新颖的，意外干扰病毒组装和功能的目标。我们发现了一个39-nt SARS-COV-2轴承结构相似性的3'-末端（干扰素 - γ激活）的结构相似性翻译的抑制剂）RNA元素先前由我们描述。我们表明谷氨酰基 - 丙酰tRNA合成酶（EPRS）结合人步态元件的蛋白质也结合了VGLE。此外，IFN-γ，一种促炎肥胖诱导的激素细胞因子和胰岛素明显增加荧光素酶报告基因的表达带有完整的VGLE。这些结果是第一个显示RNA元素在该功能后果的结果 SARS-COV-2的3'末端序列。我们假设EPR与VGLE的结合刺激了SGMRNA 表达结构和其他SARS-COV-2蛋白所需的翻译，以及用于编程的核糖体基因组复制所需的验证。我们将通过追求两个具体目标来检验这一假设：目的 1我们阐明了EPRS与SARS-COV-2 3'UTR VGLE结合在调节病毒复制中的作用和 SGMRNA翻译。在AIM 2中，我们开发针对EPRS/VGLE相互作用并阻止病毒的RNA抑制剂蛋白表达。我们预计这些基本研究将提供有关功能的第一个信息 SARS-COV-2的3'末端，将为开发用于使用的治疗剂的基础结合针对VGLE的机械不同的抗病毒剂和可能出现的 Betacoronaviruses。