Broad Spectrum Anti-viral Compounds Targeting the SKI Complex

项目摘要 RNA外切体和SKI复合体是降解宿主RNA的主要细胞机器。这 在细胞中需要机器有几个原因，包括维持当前的RNA水平 并降低细胞质RNA的水平，以便RigI和MDA5传感器可以检测到病毒 非宿主核糖核糖核酸。SKI复合体在遗传筛查中被鉴定为宿主蛋白 它与流感NS1和MERS冠状病毒ORF4a相互作用。我们能够证明这一点 在细胞中击倒这些蛋白质会导致病毒复制减少和病毒数量增加 无论病毒是否存在，干扰素都能刺激基因诱导。滑雪板 酵母中的复合体已经结晶，在对人体结构进行建模后，我们在硅胶中 确定了可能与该复合体的成员WDR61结合的化合物。单元格内 培养实验中，我们从模型中鉴定出的40种化合物中鉴定出4种化合物，即 流感病毒、MERS冠状病毒和SARS冠状病毒复制。在这项提案中，我们将确定 化合物与NS1和ORF4a对SKI络合物的作用机理。我们还将 启动体内研究以评估SKI靶向载体的抗病毒效果 流感病毒和MERS冠状病毒小鼠模型。这项工作将验证一个新的主机目标和 针对滑雪复合体的化合物，作为广泛作用的抗病毒药物。

Project Summary The RNA Exosome and SKI complex are major cellular machines that degrade host RNA. This machine is required for several reasons in the cell including maintenance of current RNA levels and to reduce the level of cytoplasmic RNA such that the RIGI and MDA5 sensors can detect viral RNA other than host RNA. The SKI complex was identified in a genetic screen as a host protein that interacts with both Influenza NS1 and MERS-CoV ORF4a. We were able to demonstrate that knocking down these proteins in cells caused a reduction in viral replication and an increase in Interferon stimulated gene induction, irrespective of whether a virus was there or not. The SKI complex in yeast has been crystalized and upon modeling of the human structure, we in silico identified compounds that could potentially bind to a member of the complex, WDR61. In cell culture experiments, we identified 4 compounds from the 40 identified in the modeling, that block Influenza virus, MERS-CoV and SARS-CoV replication. In this proposal, we will determine the mechanism of action of the compounds and NS1 and ORF4a on the SKI complex. We will also initiate in vivo studies to evaluate the antiviral effectiveness of the SKI targeted plasmids on Influenza virus and MERS-CoV mouse models. This work will validate a novel host target and compounds directed at the SKI complex as broadly acting antivirals.