Antiviral Mechanisms of 2-5A Dependent RNase L

2-5A 依赖性 RNase L 的抗病毒机制

• 批准号: 7652264
• 负责人: Robert H. Silverman
• 金额: $ 36.75万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7652264
• 关键词: Antiviral Agents; Antiviral Response; Apoptotic; Biological Assay; Cell Culture System; Cell Survival; Cells; Cleaved cell; Dominant-Negative Mutation; Double-Stranded RNA; Enzymes; Event; Fluorescence Resonance Energy Transfer; Future; GDF15 gene; Genes; Genetic Transcription; Growth; Health; Host Defense; Human; Immune; Interferon Type I; Interferons; Investigation; Knockout Mice; Knowledge; Lead; Ligase; Link; Measures; Mediator of activation protein; Mitogen-Activated Protein Kinase 3; Molecular; Mus; Pathway interactions; Pattern; Proteins; Protocols documentation; Relative (related person); Research Personnel; Ribonucleases; Role; Signal Pathway; Signal Transduction; Signaling Protein; Structure-Activity Relationship; Surface Plasmon Resonance; System; Vertebrates; Viral; Virus; Virus Replication; Wild Type Mouse; Work; base; biological adaptation to stress; cytokine; cytotoxicity; in vivo; inhibitor/antagonist; macrophage; member; neurotensin mimic 2; neutralizing antibody; novel; oligoadenylate; pathogen; programs; receptor; research study; small molecule

DESCRIPTION (provided by applicant): RNase L is a principal mediator of the innate antiviral response and is thus critically important for human health. Virus replication in higher vertebrates is restrained by interferons (IFN) that cause cells to transcribe genes encoding antiviral proteins, including the 2'-5' oligoadenylate synthetases (OAS). The viral pathogen associated molecular pattern (PAMP), double-stranded RNA (dsRNA), activates OAS to produce 5'- phosphorylated, 2', 5'-linked oligoadenylates (2-5A) whose function is to stimulate RNase L. The OAS-RNase L system thus constitutes a classical innate immune pathway that rapidly responds to PAMPs (here, dsRNA) to produce a broadly active antiviral response. In our prior studies performed under this protocol, we cloned RNase L, knocked it out in mice, established its antiviral and apoptotic activities in vivo, and determined that it induces transcriptional signaling pathways to antiviral genes. Our future GOALS are to probe fundamental events and biologic endpoints surrounding RNase L that impact on viral lifecycles. Our HYPOTHESIS is that RNase L inhibits viral replication by cleaving viral and cellular RNA, initiating a stress-response pathway and signaling expression of antiviral genes. Our Specific Aims are to: (1) optimize novel small molecular 2-5A mimics that activate RNase L and determine their effects on cell viability and viral growth; (2) determine how 2-5A activation of RNase L signals transcription of antiviral and immunoregulatory genes, including investigations into the involvement of dsRNA signaling factors and MAP kinases; and (3) determine the fundamental basis for the antiviral activity of RNase L in vivo in wild type and RNase L-null mice by measuring viral induction of type I IFNs, determining the impact of RNase L-induced IFNs on viral replication and the effects of 2-5A and 2-5A mimics on IFN induction and viral replication. The experiments described in this PROPOSAL will investigate how a major antiviral pathway is stimulated by small molecule activators of a ubiquitous and potent antiviral enzyme (RNase L). The knowledge to be gained may thus lead to strategies for controlling important human viral pathogens. Therefore, there are cogent and health-related justifications for these studies.

描述（由申请人提供）：RNase L是先天性抗病毒反应的主要介质，因此对人类健康至关重要。病毒在高等脊椎动物中的复制受到干扰素（IFN）的抑制，干扰素导致细胞转录编码抗病毒蛋白的基因，包括2 '-5'寡腺苷酸合成酶（OAS）。病毒病原体相关分子模式（PAMP），双链RNA（dsRNA），激活OAS产生5 '-磷酸化的，2'，5 '-连接的寡腺苷酸（2-5A），其功能是刺激RNA酶L。因此，OAS-RNase L系统构成了经典的先天免疫途径，其快速响应PAMP（此处为dsRNA）以产生广泛活性的抗病毒应答。在我们之前的研究中，我们克隆了RNase L，在小鼠中将其敲除，建立了其体内抗病毒和凋亡活性，并确定其诱导抗病毒基因的转录信号通路。我们未来的目标是探索影响病毒生命周期的RNase L周围的基本事件和生物学终点。我们的假设是RNase L通过切割病毒和细胞RNA，启动应激反应途径和抗病毒基因的信号表达来抑制病毒复制。我们的具体目标是：（1）优化激活RNase L的新型小分子2-5A模拟物，并确定它们对细胞活力和病毒生长的影响;（2）确定RNase L的2-5A激活如何信号传导抗病毒和免疫调节基因的转录，包括对dsRNA信号传导因子和MAP激酶的参与的研究;和（3）通过测量I型IFN的病毒诱导，确定RNase L在野生型和RNase L缺失小鼠中体内抗病毒活性的基本基础，确定RNA酶L诱导的IFN对病毒复制的影响以及2-5A和2-5A模拟物对IFN诱导和病毒复制的影响。本提案中描述的实验将研究主要的抗病毒途径如何被普遍存在且有效的抗病毒酶（RNase L）的小分子活化剂刺激。因此，所获得的知识可能会导致控制重要的人类病毒病原体的战略。因此，这些研究有令人信服的健康相关理由。