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.

概括 作为对病毒感染的先天反应的主要介体，I型干扰素（IFN-I）建立了抗病毒 通过诱导数百个干扰素的感染和未感染旁观者细胞中的状态 刺激基因（ISG）。这些ISG赋予抵抗力的协调活动的机制 对于各种病毒，调节其表达的调节回路仍然很众所周知。在职的 有了来自IFN-I反应失调的遗传性综合症患者的独特样本，我们有 确定了三十个ISG的集合，这些ISG赋予了对不同病毒的抗性（增加了RNA的保护 以及具有高病原体负担和低致病负担的DNA病毒）。我们还发现了以前未欣赏的 IFN-I信号传导的负反馈机制。我们发现这些监管电路以及ISG 表达模式在稳态和IFN-I刺激下在不同的细胞类型之间有显着变化。 现在的技术进步现在使我们能够探索这些合作的ISG抗病毒功能和负面功能 前所未有的深度和分辨率的反馈机制。 在此处提出的研究中，我们将确定ISG的子集足以对多重保护进行广泛的保护 使用新型的单细胞RNA-seq策略的病毒。这种方法提供了进行所需的吞吐量 复杂的组合实验，同时保持高分辨率以检验特定假设。结果 可能会提供新的广谱抗病毒治疗策略，这将特别有价值 新兴的病毒病原体。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烙印。这是最近描述的，但是 未完全表征的现象可能对连续感染挑战具有重要意义 和病毒敏感性。结合独特的临床样本，尖端技术和 从我们的两个实验室中的免疫病毒学多元化和补充专业知识，这些研究是可以预期的 解决长期以来的，先天抗病毒免疫的基本问题，以及开拓新方向 用于开发抗病毒疗法。