Circular RNAs in HSV1 infection and antiviral immunity

HSV1感染和抗病毒免疫中的环状RNA

• 批准号: 10495211
• 负责人: Hannah Marion Burgess
• 金额: $ 6.73万
• 依托单位: UNIVERSITY OF SURREY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-24 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10495211
• 关键词: Alleles; Antiviral Response; Binding; Biology; Cells; Chemicals; Data; Disease; Double Stranded DNA Virus; Double-Stranded RNA; Endoribonucleases; Exons; Fibroblasts; Gene Expression; Genetic Transcription; Genetic Translation; Genome; Goals; Growth; Herpes Simplex Infections; Herpesviridae; Host Defense; Human; Immediate-Early Proteins; Immunity; Infection; Innate Immune Response; Knowledge; Lead; Link; Medical; MicroRNAs; Molecular; Monitor; Pathogenicity; Pathway interactions; Pattern; Phosphotransferases; Population; Primary Infection; Protein Biosynthesis; Proteins; RNA; RNA Sequences; RNA Viruses; RNA-Binding Proteins; Regulator Genes; Reproduction; Ribonucleases; Role; Shapes; Small RNA; Testing; Viral; Viral Proteins; Virion; Virus; Virus Diseases; Virus Replication; Work; antiviral immunity; base; circular RNA; design; human pathogen; inhibitor; mRNA Decay; mutant; novel; pathogen; preservation; prevent; protein protein interaction; response; transcriptome sequencing

Project Summary Circular (circ) RNAs are a recently described class of RNAs generated by backsplicing of adjacent exons, whose expression is altered in several disease states and some of the few viral infections in which they have been studied. Functionally, circRNAs rarely direct protein synthesis but have been linked to the sequestration of microRNAs and RNA binding proteins (RNA-BPs). CircRNAs were recently implicated in controlling the activation of double-stranded (ds) RNA-activated eIF2α kinase PKR by providing an abundant pool of inhibitory short duplex RNA sequences. Bound inactive PKR can be rapidly mobilized by if RNase L is activated, resulting in endoribonucleolytic cleavage of circRNAs. Generated by both RNA virus replication and dsDNA virus transcription from opposing genome strands, dsRNA is a potent pathogen associated molecular pattern (PAMP) whose recognition by PKR and the OAS/RNase L pathway leads to an abrupt defensive shutdown of mRNA translation to thwart viral protein synthesis. In addition to possible virus and circRNA -specific interactions, the potential of circRNAs to calibrate dsRNA responses therefore suggests a broad significance to virus biology. However, precisely how the infected cell circRNA population responds to herpes simplex type 1 (HSV1) infection and impacts virus reproduction remains unknown and represents a significant knowledge gap. The overall goal of this proposal is to understand how circRNAs contribute to HSV1 infection and antiviral immunity. Our preliminary data in HSV1-infected cells indicate that circRNAs are protected from degradation by the viral endoribonuclease vhs and are in fact selectively upregulated through a mechanism dependent on the viral immediate early protein ICP27. We also find that PKR activation is dependent on RNase L function, suggesting that circRNAs are important inhibitors of PKR in HSV1 infection that must be degraded for a full antiviral response. These data are consistent with the novel hypothesis that HSV1 actively manipulates this class of host RNAs to evade cell intrinsic antiviral defenses and preserve protein synthesis and viral replication. This proposal will be divided into two aims to fully understand 1) how HSV1 shapes the infected cell circRNAome and 2) the functional impact of these alterations. These studies will reveal how circRNAs are manipulated by HSV1 and their importance in infection and immunity to a medically important human pathogen. Ultimately, this work could lead to new strategies for treating a wide variety of viral infections and have significant implications for our understanding of infection, host defenses and circRNA biology.

项目概要 环状 (circ) RNA 是最近描述的一类通过相邻外显子反向剪接生成的 RNA， 它们的表达在几种疾病状态和一些少数病毒感染中发生改变 被研究过。从功能上讲，circRNA很少指导蛋白质合成，但与隔离有关 microRNA 和 RNA 结合蛋白 (RNA-BP)。 CircRNA 最近被认为参与控制 通过提供丰富的抑制库来激活双链 (ds) RNA 激活的 eIF2α 激酶 PKR 短双链体 RNA 序列。如果 RNase L 被激活，结合的非活性 PKR 可以被快速动员， 导致 circRNA 的核糖核酸内切裂解。由RNA病毒复制和双链DNA产生 病毒从相反的基因组链转录，dsRNA 是一种有效的病原体相关分子模式 (PAMP)，其被 PKR 和 OAS/RNase L 通路识别导致突然的防御性关闭 mRNA 翻译阻止病毒蛋白质合成。除了可能的病毒和circRNA特异性 因此，circRNA 校准 dsRNA 反应的潜力表明其具有广泛的意义 病毒生物学。然而，受感染的细胞 circRNA 群体究竟如何对 1 型单纯疱疹做出反应 (HSV1) 感染及其对病毒繁殖的影响仍然未知，存在重大的知识差距。 该提案的总体目标是了解 circRNA 如何促进 HSV1 感染和 抗病毒免疫力。我们在 HSV1 感染细胞中的初步数据表明 circRNA 受到保护 由病毒核糖核酸内切酶 vhs 降解，实际上通过一种机制选择性上调 依赖于病毒立即早期蛋白 ICP27。我们还发现 PKR 激活取决于 RNase L 功能，表明 circRNA 是 HSV1 感染中 PKR 的重要抑制剂，必须 降解以获得完全的抗病毒反应。这些数据与 HSV1 主动 操纵这类宿主 RNA 来逃避细胞内在的抗病毒防御并保护蛋白质合成 和病毒复制。该提案将分为两个目标，以充分理解 1）HSV1 如何塑造 受感染细胞的circRNAome和2）这些改变的功能影响。这些研究将揭示如何 circRNA 由 HSV1 操纵，及其在医学上重要的感染和免疫中的重要性 人类病原体。最终，这项工作可能会带来治疗多种病毒感染的新策略 对我们理解感染、宿主防御和 circRNA 生物学具有重要意义。