Roles for hepatitis C virus-derived circular RNAs in infected cells

丙型肝炎病毒衍生的环状 RNA 在受感染细胞中的作用

• 批准号: 10442607
• 负责人: PETER SARNOW
• 金额: $ 19.68万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10442607
• 关键词: Address; Affect; Amber; Antiviral Agents; Antiviral Response; Autologous; Back; Binding Sites; Biochemical; Bioinformatics; Biological; Biological Assay; Biological Markers; C-terminal; CRISPR/Cas technology; Catalytic Domain; Cell Nucleus; Cells; Cirrhosis; Code; Cytoplasm; Differentiation and Growth; Disease; Ectopic Expression; Endoplasmic Reticulum; Eukaryotic Cell; Exonuclease; Flaviviridae; Fluorescent in Situ Hybridization; Gene Expression; Generations; Genetic; Genetic Transcription; Genome; Goals; Growth; Hepatitis C virus; Hepatocyte; Human; Immune response; Immunoprecipitation; Individual; Infection; Internal Ribosome Entry Site; Kinetics; Label; Laboratories; Lead; Ligase; Liver; Malignant - descriptor; Mass Spectrum Analysis; Mission; Monitor; Movement; Nuclear; Nuclear RNA; Nucleotides; Oligonucleotides; Open Reading Frames; Organoids; Outcome; Pathogenesis; Patients; Peptide Hydrolases; Peptides; Persons; Polymerase; Predisposition; Process; Production; Proteins; Public Health; RNA; RNA Splicing; RNA Viruses; RNA replication; Research; Resistance; Ribosomal RNA; Ribosomes; Rice; Role; Small Interfering RNA; Terminator Codon; Testing; Transfer RNA; Translations; United States National Institutes of Health; Viral; Virus; Virus Diseases; cell growth; circular RNA; combat; endonuclease; endoplasmic reticulum stress; gene function; genomic RNA; inhibitor; innovation; mRNA Precursor; member; migration; novel; novel virus; patient population; polypeptide; prevent; response; tRNA Ligase; viral RNA

While novel direct-acting antivirals (DAA) against hepatitis C virus (HCV), such as anti-polymerase and anti- proteinase compounds are effective in many patients, these treatments are still very expensive and not available to most of the 170 million people world-wide infected with HCV. In addition, it is not known whether every patient population will respond to the current DAA treatments. Thus, it is significant to continue to search for new compounds that target both HCV and viral host susceptibility factors to combat virus infection. An astonishing recent discovery made by our laboratory revealed that the HCV genomic RNA is processed during viral infection to yield hundreds of different virus-derived circular RNAs (circRNAs). These circRNAs are generated from all parts of the 10,000-nucleotide viral RNA genome, including circRNAs that contain the viral internal ribosome entry site (IRES). It is hypothesized that these circRNAs present a novel class of viral RNA species that likely display novel functions in infected and uninfected bystander cells. Because these viral circRNAs are predicted to be long-lived they could also have important roles as biomarkers. Thus, exploring the pro- and anti-viral effects of the HCV-derived circular RNAs is a significant venue of research and will point to new Achilles’ heels in RNA viruses. The long-term goals of this application are to explore the roles for the identified viral circRNAs in pro- and anti-viral responses. Two specific aims are proposed to accomplish these goals. First, the mechanism by which the viral circRNAs are generated is being investigated. The hypothesis is that HCV subverts a cellular splicing mechanism that modulates the unfolded protein response in the endoplasmic reticulum. Secondly, it will be examined whether highly abundant circRNAs and IRES-containing circRNAs have functional roles on HCV gene expression in infected cells. The functions of IRES-circRNA translation products in the viral infectious cycle will be examined both in cultured liver cells and human liver organoids that can be infected with autologous HCV. The expected outcomes of this application will address fundamental aspects about the functions of novel circular HCV-derived RNAs in the viral infectious cycle. This proposed research is innovative because it will examine whether novel virus-derived circRNAs can be targeted in antiviral approaches in RNA virus-infected cells.

虽然针对丙型肝炎病毒 (HCV) 的新型直接作用抗病毒药物 (DAA)，例如抗聚合酶和抗蛋白酶化合物对许多患者有效，但这些治疗方法仍然非常昂贵，并且全世界 1.7 亿 HCV 感染者中的大多数人无法获得这些治疗方法。此外，尚不清楚是否每个患者群体都会对当前的 DAA 治疗产生反应。因此，继续寻找同时针对HCV和病毒宿主易感因子的新化合物来对抗病毒感染具有重要意义。我们实验室最近取得的一项惊人发现表明，HCV 基因组 RNA 在病毒感染过程中被加工，产生数百种不同的病毒衍生环状 RNA (circRNA)。这些 circRNA 由 10,000 个核苷酸的病毒 RNA 基因组的所有部分生成，包括含有病毒内部核糖体进入位点 (IRES) 的 circRNA。据推测，这些 circRNA 代表了一类新型病毒 RNA 种类，它们可能在受感染和未受感染的旁观细胞中表现出新的功能。由于这些病毒 circRNA 预计寿命较长，因此它们也可以作为生物标志物发挥重要作用。因此，探索 HCV 来源的环状 RNA 的促病毒和抗病毒作用是一个重要的研究领域，并将指出 RNA 病毒的新致命弱点。该应用的长期目标是探索已识别的病毒 circRNA 在促病毒和抗病毒反应中的作用。为了实现这些目标，提出了两个具体目标。首先，正在研究病毒circRNA的产生机制。该假设认为，HCV 颠覆了调节内质网中未折叠蛋白反应的细胞剪接机制。其次，将检查高丰度的 circRNA 和含有 IRES 的 circRNA 是否对感染细胞中的 HCV 基因表达具有功能作用。 IRES-circRNA 翻译产物在病毒感染周期中的功能将在培养的肝细胞和可感染自体 HCV 的人类肝类器官中进行检查。该应用的预期结果将解决新型环状 HCV 衍生 RNA 在病毒感染周期中的功能的基本问题。这项拟议的研究具有创新性，因为它将检验新型病毒衍生的 circRNA 是否可以作为 RNA 病毒感染细胞的抗病毒方法的靶标。

项目成果

Virus-derived circular RNAs populate hepatitis C virus-infected cells.

• DOI: 10.1073/pnas.2313002121
• 发表时间: 2024-02
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Qian M Cao;Pakpoom Boonchuen;Tzu-Chun Chen;Shaohua Lei;Kunlaya Somboonwiwat;P. Sarnow