Structure-function determination of the Hepatitis C Virus negative RNA strand???s

丙型肝炎病毒阴性 RNA 链的结构-功能测定

• 批准号: 8524889
• 负责人: Edward Anhoa Pham
• 金额: $ 3.23万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-24 至 2016-06-23
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8524889
• 关键词: 5' Untranslated Regions; Acylation; Adverse effects; Affect; Antiviral Agents; Beryllium; Binding; Biological; Biological Process; Cell Culture Techniques; Cells; Crystallography; Data; Elements; Foundations; Future; Genetic Transcription; Genome; Genomics; Goals; Gold; Hepatitis C virus; Hydroxyl Radical; Indium; Induced Mutation; Infection; Information Storage; Lead; Life Cycle Stages; Liver; Malignant neoplasm of liver; Maps; Mediating; Methods; MicroRNAs; Microfluidics; Modeling; Molecular Virology; Monitor; Mutation; Names; Nuclear Magnetic Resonance; Nucleic Acid Regulatory Sequences; Patients; Pegylated Interferon Alfa; Play; Polymerase; Primer Extension; Proteins; Quartz; RNA; RNA Binding; RNA Viruses; RNA chemical synthesis; Regulatory Element; Research; Resistance; Role; Site; Structure; Testing; Time; Translations; United States; Viral; Viral Proteins; Virus; Virus Replication; anti-hepatitis C; base; design; effective therapy; fluorophore; inhibitor/antagonist; liver transplantation; mutant; next generation; novel; public health relevance; screening; small molecule; standard of care; therapeutic target; tool; viral RNA

DESCRIPTION (provided by applicant): Over 150 million people are infected with hepatitis C virus (HCV) worldwide. Current standard of care (SOC) based on the foundation of pegylated alpha interferon is inadequate for many patients and associated with significant side effects. A better understanding of the molecular virology of HCV can lead to better rational design for newer and more effective therapies. So far, viral proteins have been the predominant focus of therapeutic targets. However, research has shown that the HCV RNA itself plays many important roles in the viral life cycle, beyond information storage. Specifically, the 5' untranslated region (UTR) of the HCV RNA and the corresponding region on the complementary negative strand, called the 3' term (-) - which is the site for initiation of progeny plus strand genomes - contain distinct RNA elements that are important for their regulatory functions. Using a novel RNA secondary structure mapping tool named sf-SHAPE (single fluorophore selective 2'-hydroxyl acylation analyzed by primer extension), I have determined the secondary structure of the HCV 5' UTR and have also shown that the 5' UTR secondary structure is altered by the presence of the miR-122, an abundant, liver-specific, micro RNA that has been shown to be required for HCV replication. However, much less is known about the structural details of the 3' term (-) and how they relate to this region's critical functions in th virus's life cycle. Since sf-SHAPE allows for rapid and accurate determination of RNA secondary structures, I propose here to study the RNA secondary structure of the HCV 3' term (-) in multiple biological contexts. Such information is critical to a better understanding of the HCV lif cycle, in aiding the rational design of potential next-generation anti-HCV therapies, and for future screening for inhibitors that might purposefully target essential RNA elements in the 3' term (-), thereby providing proof- of-concept for a potential novel class of RNA-targeting small molecules with application to HCV and RNA viruses in general.

描述（由申请人提供）：全世界有超过1.5亿人感染丙型肝炎病毒（HCV）。目前基于聚乙二醇化α干扰素基础的标准治疗（SOC）对许多患者来说是不够的，并伴有显著的副作用。更好地了解HCV的分子病毒学可以更好地合理设计更新和更有效的治疗方法。到目前为止，病毒蛋白一直是治疗靶点的主要焦点。然而，研究表明，HCV RNA本身在病毒生命周期中发挥着许多重要作用，而不仅仅是信息存储。具体地说，HCV RNA的5'非翻译区（UTR）和互补负链上的相应区域，称为3'端（-），这是子代起始的位点 正链基因组-包含对其调节功能重要的不同RNA元件。使用一种名为sf-SHAPE（通过引物延伸分析的单荧光团选择性2 '-羟基酰化）的新型RNA二级结构作图工具，我已经确定了HCV 5' UTR的二级结构，并且还表明5' UTR二级结构因miR-122的存在而改变，miR-122是一种丰富的肝脏特异性微小RNA，已被证明是HCV复制所需的。然而，关于3'端（-）的结构细节以及它们如何与该区域在病毒生命周期中的关键功能相关的知之甚少。由于sf-SHAPE可以快速准确地测定RNA二级结构，因此我建议在多种生物学背景下研究HCV 3'端（-）的RNA二级结构。这样的信息对于更好地理解HCV生命周期、帮助合理设计潜在的下一代抗HCV疗法以及对于将来筛选可能有目的地靶向3'端（-）中的必需RNA元件的抑制剂是至关重要的，从而为潜在的新型RNA靶向小分子提供概念验证，其通常应用于HCV和RNA病毒。