Roles for microRNA-122 and circular RNAs in flavivirus RNA amplification

microRNA-122 和环状 RNA 在黄病毒 RNA 扩增中的作用

• 批准号: 9912689
• 负责人: PETER SARNOW
• 金额: $ 54.95万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-15 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9912689
• 关键词: Acylation; Address; Adenosine; Affect; Antiviral Agents; Base Pairing; Binding Sites; Biochemical Genetics; Bioinformatics; Biological Assay; Biotinylation; Cells; Cirrhosis; Complex; Cytoplasm; DNA; DNA-Directed RNA Polymerase; Dengue Virus; Detection; Dicer Enzyme; Disease; Escape Mutant; Exonuclease; Flaviviridae; Flavivirus; GTP-Binding Protein alpha Subunits, Gs; Gene Expression Regulation; Genes; Genetic; Genome; Goals; Growth; Hepatitis C; Hepatitis C virus; Hepatocyte; Herpesviridae; Hydroxyl Radical; Individual; Infection; Lead; Life Cycle Stages; Liver; Maintenance; Malignant - descriptor; Mediating; Methodology; Methylation; MicroRNAs; Mission; Modification; Monitor; Mutation; Nucleic Acids; Nucleotides; Occupations; Oligonucleotides; Outcome; Pathogenesis; Pathway interactions; Patients; Peptides; Phase II Clinical Trials; Primer Extension; Process; Property; Proteins; Proteomics; Psoralens; Public Health; RNA; RNA amplification; RNA analysis; RNA methylation; RNA-Protein Interaction; Reader; Reagent; Research; Resistance; Resolution; Role; Serum; Site; Structure; Translating; Translation Initiation; Translations; United States National Institutes of Health; Untranslated RNA; Vaccines; Variant; Viral; Viral Genome; Viral Pathogenesis; Viral Physiology; Viral Proteins; Virus; West Nile virus; Zika Virus; base; circular RNA; genetic approach; innovation; member; mutant; novel; prevent; protein complex; tool; viral RNA; virus host interaction

New direct-acting antivirals against hepatitis C virus (HCV) cure the virus in most patients. However, these compounds are expensive and not available to most of the 170 million people that are worldwide infected with HCV. There are also no approved vaccines or anti-viral compounds for other members of the flaviviridae, such as Dengue virus, West Nile virus or Zika virus. Thus, identification of cellular genes that are essential for virus propagation is highly significant, because such targets are not regulated by the error-prone viral RNA polymerase and, therefore, offer a high barrier to resistance. The long-term goal of this application is to explore the mechanism by which certain noncoding RNAs, such as microRNAs and circular RNAs, display pro-viral or anti-viral activities. It is known that microRNA miR-122 attaches as an oligomeric complex to the 5’ end of the viral genome and protects it from degradation by host exonucleases. Curiously, escape mutants that harbor a single C3U mutation in the viral genome were detected in the serum of 122-antagomir treated patients. The overall objective of the first aim to is identify interactions of between C3U HCV genome with nucleic acids or proteins that allow the virus to persist when miR-122 abundance is reduced. The central hypothesis is that novel RNA-RNA or protein RNA interactions in the C3U HCV genome allow stabilization and expression of the viral genome in the cells during reduced miR-122 abundances. Novel cell-based protein biotinylation assays and approaches that detect tertiary RNA structures will be used to study RNA-protein and RNA-RNA interactions in wildtype and mutant viral RNAs in cultured liver cells. Steps in the viral life cycle that are modulated by such nucleic acid-protein interactions will be identified. The overall objective of the second aim is based upon the finding that the host cell-derived circular RNA (cRNAs) landscape is altered during HCV infection. The potential mechanisms by which cRNAs exert their pro- and antiviral functions will be explored. First, effects of cRNA-mediated sequestration of proteins and microRNAs on HCV RNA amplification will be examined. Because methylation of specific adenosines in cRNAs has been shown to induce translation initiation in cRNAs, the methylation status in cRNAs will be determined. The properties of methylated cRNAs to synthesize small peptides will be examined using genetic and proteomic approaches. Overall, the application details innovative paradigm-shifting concepts to study roles for noncoding RNAs in virus-host interactions, using novel detection methodologies. The rationale for this proposal is that noncoding RNAs, such as microRNAs and cRNAs, affect HCV pathogenesis. It has been shown that microRNAs can be targeted in HCV patients, resulting in loss of viral RNA abundance. Thus, targeting pro-viral cRNAs in the liver offers a novel antiviral strategy.

针对丙型肝炎病毒（HCV）的新直接作用抗病毒药在大多数患者中治愈该病毒。但是，这些 化合物很昂贵，而且在全球感染的1.7亿人中，大多数人都无法使用 HCV。也没有针对黄椎科动物的其他成员的批准疫苗或抗病毒化合物 作为登革热病毒，西尼罗河病毒或寨卡病毒。那是对病毒必不可少的细胞基因的鉴定 传播非常重要，因为此类靶标不受易易受错误的病毒RNA的调节 聚合酶，因此为抗性提供了很高的障碍。该应用程序的长期目标是探索 某些非编码RNA（例如microRNA和圆形RNA）的机制显示促病毒或 反病毒活动。众所周知，microRNA mir-122作为低聚复合物附着在5'的5'端 病毒基因组并保护其免受宿主外切酶的降解。奇怪的是，逃脱了藏有一个的突变体 在122-塔哥哥治疗的患者的血清中检测到病毒基因组中的单个C3U突变。 第一个目的的总体目标是确定C3U HCV基因组与核酸或 当miR-122抽象降低时，允许病毒持续存在的蛋白质。中心假设是 C3U HCV基因组中的新型RNA-RNA或蛋白RNA相互作用允许稳定和表达 在减少的miR-122丰度中，细胞中的病毒基因组。新型细胞基蛋白生物素化测定 以及检测三级RNA结构的方法将用于研究RNA-蛋白质和RNA-RNA 在培养的肝细胞中野生型和突变病毒RNA中的相互作用。病毒生命周期的步骤 通过这种核酸蛋白相互作用调节。第二个目标的总体目标是 基于发现在HCV期间改变了宿主细胞衍生的圆形RNA（CRNA）景观 感染。将探索CRNA发挥促病毒功能的潜在机制。 首先，CRNA介导的蛋白质和microRNA对HCV RNA扩增的影响将是 检查。因为已显示CRNA中特定腺苷的甲基化已诱导翻译 在CRNA中的启动，将确定CRNA中的甲基化状态。甲基化crnas的特性 将使用遗传和蛋白质组学方法检查综合小宠物。总体而言，应用程序 详细介绍创新的范式转移概念，以研究非编码RNA在病毒宿主相互作用中的作用， 使用新颖的检测方法。该提议的理由是未编码的RNA，例如 microRNA和CRNA，影响HCV发病机理。已经显示MicroRNA可以靶向HCV 患者，导致病毒RNA丰度丧失。那是针对肝脏中的促病毒CRNA提供的小说 抗病毒策略。