Next Generation Resolution of Antiviral Gene Networks

抗病毒基因网络的下一代解决方案

• 批准号: 10461962
• 负责人: Dusan Bogunovic
• 金额: $ 66.2万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-22 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10461962
• 关键词: Address; Antiviral Agents; Biological Assay; COVID-19 outbreak; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Communicable Diseases; Communication; Complex; Cytometry; DNA Viruses; Data; Epithelial Cells; Feedback; Fibroblasts; Gene Expression Profile; Genes; Growth; Herpesvirus 1; Host Defense; Human; ISG15 gene; Immune; Individual; Infection; Influenza A virus; Inherited; Interferon Type I; Interferons; JAK1 gene; Journals; Laboratories; Laboratory Study; Mediating; Mediator of activation protein; Memory; Nature; Pathogenicity; Peripheral Blood Mononuclear Cell; Phenotype; Physiological; Predisposition; Proteins; RNA Viruses; Refractory; Regulation; Resistance; Resolution; Sampling; Secondary to; Signal Transduction; Syndrome; System; Techniques; Technology; Testing; Therapeutic; Time; Toxic effect; Vaccination; Vesicular stomatitis Indiana virus; Viral; Virus; Virus Diseases; Work; Yellow Fever; Zika Virus; airway epithelium; antiviral drug development; antiviral immunity; cell type; combinatorial; experimental study; gene function; gene network; high dimensionality; human model; imprint; in vitro Model; in vivo; new technology; next generation; novel; pandemic disease; pathogenic virus; programs; respiratory pathogen; response; side effect; single-cell RNA sequencing

SUMMARY As the primary mediators of the innate response to viral infection, type I interferons (IFN-I) establish an antiviral state in both infected and uninfected bystander cells through the induction of several hundreds of interferon stimulated genes (ISGs). The mechanisms by which the coordinated activities of these ISGs confer resistance to diverse viruses, and the regulatory circuits that modulate their expression remain poorly understood. Working with unique samples from individuals with hereditary syndromes of dysregulated IFN-I responsiveness, we have identified a collection of thirty ISGs that confer resistance to diverse viruses (increased protection against RNA and DNA viruses with both high and low pathogenic burden). We have also uncovered previously unappreciated negative feedback mechanisms of IFN-I signaling. We have found that these regulatory circuits, as well as ISG expression patterns, vary significantly across different cell types at steady state and upon IFN-I stimulation. Recent technological advances now enable us to explore these cooperative ISG antiviral functions and negative feedback mechanisms at unprecedented depth and resolution. In the studies proposed here, we will identify subsets of ISGs sufficient to confer broad protection against multiple viruses using a novel single cell RNA-Seq strategy. This approach provides the throughput required to conduct complex combinatorial experiments while maintaining the high resolution to test specific hypotheses. Results may offer new broad spectrum antiviral therapeutic strategies, which would be of particular value against emerging viral pathogens. We will also investigate in detail the mechanisms by which IFN-I signaling establishes a lasting imprint on cellular responsiveness to subsequent IFN-I stimulation. This recently described but incompletely characterized phenomenon likely has important implications for successive infectious challenges and viral susceptibilities. Combining a unique collection of clinical samples, cutting edge technologies, and diverse and complementary expertise in immunovirology from our two laboratories, these studies are expected to address long standing, fundamental questions in innate antiviral immunity, as well as to pioneer new directions for developing antiviral therapies.

总结 作为对病毒感染的先天性应答的主要介质，I型干扰素（IFN-I）建立了抗病毒的免疫调节系统。 通过诱导数百种干扰素， 刺激基因（ISG）。这些ISG的协调活动赋予抗性的机制 不同的病毒，以及调节其表达的调控电路仍然知之甚少。工作 通过从IFN-I反应性失调的遗传综合征患者中提取的独特样本，我们 鉴定了30个ISG的集合，这些ISG赋予对不同病毒的抗性（增加对RNA的保护 和具有高和低致病性负荷的DNA病毒）。我们还发现了以前未被重视的 IFN-I信号传导的负反馈机制。我们发现，这些调节电路，以及ISG 表达模式在稳态和IFN-1刺激后在不同细胞类型中显著变化。 最近的技术进步现在使我们能够探索这些合作ISG抗病毒功能和负作用。 以前所未有的深度和分辨率提供反馈机制。 在这里提出的研究中，我们将确定足以提供广泛保护的ISG子集， 病毒使用一种新的单细胞RNA-Seq策略。此方法提供了进行以下操作所需的吞吐量 复杂的组合实验，同时保持高分辨率，以测试特定的假设。结果 可能提供新的广谱抗病毒治疗策略， 新出现的病毒病原体。我们还将详细研究IFN-I信号建立的机制， 对随后的IFN-I刺激的细胞反应性的持久印记。这是最近描述的，但 不完全表征的现象可能对连续的感染性挑战具有重要意义 和病毒感染性。结合独特的临床样本收集，尖端技术， 我们的两个实验室在免疫病毒学方面具有多样和互补的专业知识，预计这些研究 解决先天性抗病毒免疫中长期存在的基本问题，并开拓新的方向 用于开发抗病毒疗法。