宿主激酶在宿主细胞周期、先天免疫信号传导、对病毒感染的应激反应和炎症中发挥至关重要的作用。以往的研究表明，冠状病毒特异性地靶向激酶级联以逆转宿主细胞对感染的反应，并依赖于宿主激酶活性对病毒蛋白进行磷酸化以增强复制。鉴于已经获得FDA批准用于治疗肿瘤、纤维化和其他人类疾病的激酶抑制剂数量众多，它们成为对抗新型冠状病毒感染的一类有吸引力的化合物。为了进一步了解β冠状病毒感染对宿主激酶组的响应，我们采用多重抑制性珠质色谱质谱技术（MIB-MS），分别对中东呼吸综合征冠状病毒（MERS-CoV）和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）感染的人肺上皮细胞系进行了分析。我们的MIB-MS分析揭示了MERS-CoV感染后mTOR和MAPK信号传导级联的激活，以及SARS-CoV-2感染后MAPK、PI3K和mTOR信号传导的激活。通过化学基因组筛选，我们发现临床相关的PI3K/mTOR抑制剂以纳摩尔浓度能够抑制冠状病毒的复制，类似于直接作用的抗病毒药物。该研究为鉴定广谱的宿主靶向治疗手段以减少β冠状病毒的复制奠定了基础，并可在未来的疫情爆发和流行中快速进行再利用。本研究产生的蛋白质组学、磷酸蛋白组学和MIB-MS数据可在Proteomics Identification Database（PRIDE）资料库项目标识符PXD040897和PXD040901下获得。

Host kinases play essential roles in the host cell cycle, innate immune signaling, the stress response to viral infection, and inflammation. Previous work has demonstrated that coronaviruses specifically target kinase cascades to subvert host cell responses to infection and rely upon host kinase activity to phosphorylate viral proteins to enhance replication. Given the number of kinase inhibitors that are already FDA approved to treat cancers, fibrosis, and other human disease, they represent an attractive class of compounds to repurpose for host-targeted therapies against emerging coronavirus infections. To further understand the host kinome response to betacoronavirus infection, we employed multiplex inhibitory bead mass spectrometry (MIB-MS) following MERS-CoV and SARS-CoV-2 infection of human lung epithelial cell lines. Our MIB-MS analyses revealed activation of mTOR and MAPK signaling following MERS-CoV and SARS-CoV-2 infection, respectively. SARS-CoV-2 host kinome responses were further characterized using paired phosphoproteomics, which identified activation of MAPK, PI3K, and mTOR signaling. Through chemogenomic screening, we found that clinically relevant PI3K/mTOR inhibitors were able to inhibit coronavirus replication at nanomolar concentrations similar to direct-acting antivirals. This study lays the groundwork for identifying broad-acting, host-targeted therapies to reduce betacoronavirus replication that can be rapidly repurposed during future outbreaks and epidemics. The proteomics, phosphoproteomics, and MIB-MS datasets generated in this study are available in the Proteomics Identification Database (PRIDE) repository under project identifiers PXD040897 and PXD040901.