Next Generation Resolution of Antiviral Gene Networks

摘要 作为对病毒感染的先天反应的主要介质，I型干扰素(IFN-I)建立了一种抗病毒 通过诱导数百种干扰素在感染和未感染的旁观者细胞中的状态 刺激基因(ISG)。这些ISG的协调活动产生抗性的机制 对不同的病毒，以及调节它们表达的调控电路仍然知之甚少。劳作 通过来自患有干扰素-I反应失调遗传综合征的个体的独特样本，我们有 确定了一个由30个ISG组成的集合，这些ISG具有对各种病毒的抵抗力(增强了对RNA的保护 以及具有高和低致病负担的DNA病毒)。我们还发现了以前未被赏识的 干扰素-I信号的负反馈机制。我们发现，这些调节电路以及ISG 在稳定状态下和在干扰素-I刺激下，不同类型的细胞的表达模式有显著差异。 最近的技术进步现在使我们能够探索这些合作的ISG抗病毒功能和阴性 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ISG的子集足以提供广泛的保护，以抵御多重 使用新的单细胞RNA-SEQ策略的病毒。此方法提供了执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，以检验特定的假设。结果 可能会提供新的广谱抗病毒治疗策略，这将对预防 新出现的病毒病原体。我们还将详细研究干扰素-I信号建立的机制 对随后的干扰素-I刺激的细胞反应性的持久印记。这是最近描述的，但 不完全描述的现象可能对连续的传染性挑战具有重要影响 以及对病毒的易感性。结合了独特的临床样本集合、尖端技术和 我们两个实验室在免疫病毒学方面的不同和互补的专业知识，这些研究是可望进行的 解决先天抗病毒免疫中长期存在的基本问题，以及开拓新的方向 用于开发抗病毒疗法。

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.