Broad Spectrum Anti-viral Compounds Targeting the SKI Complex

项目摘要 RNA外泌体和SKI复合物是降解宿主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，使得RIGI和MDA 5传感器可以检测病毒 RNA不是宿主RNA。SKI复合物在遗传筛选中被鉴定为宿主蛋白 与流感病毒NS 1和MERS-CoV ORF 4a相互作用。我们能够证明 在细胞中敲低这些蛋白质会导致病毒复制减少， 干扰素刺激基因诱导，不管是否有病毒存在。滑雪 酵母中的复合物已经结晶，在模拟人体结构后，我们在计算机上 鉴定了可能与复合物WDR 61的一个成员结合的化合物。在细胞 培养实验中，我们从建模中确定的40种化合物中确定了4种化合物， 流感病毒、MERS-CoV和SARS-CoV复制。在本建议中，我们将确定 化合物和NS 1和ORF 4a对SKI复合物的作用机制。我们还将 启动体内研究，以评估SKI靶向质粒对 流感病毒和MERS-CoV小鼠模型。这项工作将验证一种新的宿主靶标， 针对SKI复合物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.