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

基本信息

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
项目类别：
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

项目摘要

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.

新型直接作用的抗丙型肝炎病毒（HCV）抗病毒药物可以治愈大多数患者的病毒。化合物价格昂贵，全球 1.7 亿感染者中的大多数人都无法获得 HCV 也没有批准用于黄病毒科其他成员的疫苗或抗病毒化合物，例如例如登革热病毒、西尼罗河病毒或寨卡病毒，因此，鉴定病毒所必需的细胞基因。传播非常重要，因为这些目标不受容易出错的病毒 RNA 的调节聚合酶，因此提供了很高的耐药性屏障。该应用的长期目标是探索。某些非编码 RNA（例如 microRNA 和环状 RNA）显示前病毒或病毒的机制已知 microRNA miR-122 作为寡聚复合物附着在 5' 端。奇怪的是，病毒基因组并保护其免受宿主核酸外切酶的降解。在 122-antagomir 治疗患者的血清中检测到病毒基因组中的单个 C3U 突变。第一个目标的总体目标是确定 C3U HCV 基因组与核酸之间的相互作用或当 miR-122 丰度减少时，病毒能够持续存在的蛋白质核心假设是： C3U HCV 基因组中新颖的 RNA-RNA 或蛋白质 RNA 相互作用允许稳定和表达新型基于细胞的蛋白质生物素化测定期间细胞中的病毒基因组。检测RNA三级结构的方法将用于研究RNA-蛋白质和RNA-RNA 培养的肝细胞中野生型和突变型病毒 RNA 的相互作用。第二个目标的总体目标是确定这种核酸-蛋白质相互作用的调节。基于 HCV 期间宿主细胞来源的环状 RNA (cRNA) 景观发生改变的发现我们将探讨 cRNA 发挥其促病毒和抗病毒功能的潜在机制。首先，cRNA 介导的蛋白质和 microRNA 隔离对 HCV RNA 扩增的影响将是因为 cRNA 中特定腺苷的甲基化已被证明可以诱导翻译。 cRNA 中的起始，cRNA 中的甲基化状态将被确定。总体而言，将使用遗传和蛋白质组学方法来检查合成小肽的应用。详细介绍了研究非编码 RNA 在病毒-宿主相互作用中的作用的创新范式转换概念，使用新颖的检测方法这一提议的基本原理是非编码RNA，例如 microRNA 和 cRNA 影响 HCV 发病机制已经表明 microRNA 可以靶向 HCV。因此，针对肝脏中的前病毒 cRNA 提供了一种新的方法。抗病毒策略。