Wu Kan Xing, Yogarajah Thinesshwary, Loe Marcus Wing Choy, Kaur Parveen, Lee Regina Ching Hua, Mok Chee Keng, Wong Yi Hao, Phuektes Patchara, Yeo Li Sze, Chow Vincent T K, Tan Yong Wah, Chu Justin Jang Hann
Lee Kong Chian School of Medicine, Nanyang Technological University, Singapore 308232, Singapore.
Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore 117597, Singapore.
Acta Pharm Sin B. 2023 May;13(5):2039-2055. doi: 10.1016/j.apsb.2023.03.015. Epub 2023 Mar 18.
Positive-sense RNA viruses modify intracellular calcium stores, endoplasmic reticulum and Golgi apparatus (Golgi) to generate membranous replication organelles known as viral factories. Viral factories provide a conducive and substantial enclave for essential virus replication concentrating necessary cellular factors and viral proteins in proximity. Here, we identified the vital role of a broad-spectrum antiviral, peruvoside in limiting the formation of viral factories. Mechanistically, we revealed the pleiotropic cellular effect of Src and PLC kinase signaling cyclin-dependent kinase 1 signaling leads to Golgi-specific brefeldin A-resistance guanine nucleotide exchange factor 1 (GBF1) phosphorylation and Golgi vesiculation by peruvoside treatment. The ramification of GBF1 phosphorylation fosters GBF1 deprivation consequentially activating downstream antiviral signaling by dampening viral factories formation. Further investigation showed signaling of ERK1/2 pathway cyclin-dependent kinase 1 activation leading to GBF1 phosphorylation at Threonine 1337 (T1337). We also showed 100% of protection in peruvoside-treated mouse model with a significant reduction in viral titre and without measurable cytotoxicity in serum. These findings highlight the importance of dissecting the broad-spectrum antiviral therapeutics mechanism and pave the way for consideration of peruvoside, host-directed antivirals for positive-sense RNA virus-mediated disease, in the interim where no vaccine is available.
正链RNA病毒会修饰细胞内的钙储存、内质网和高尔基体，以生成被称为病毒工厂的膜性复制细胞器。病毒工厂为病毒的关键复制提供了一个有利且充实的区域，它将必要的细胞因子和病毒蛋白聚集在附近。在此，我们确定了一种广谱抗病毒药物——黄夹次苷在限制病毒工厂形成方面的重要作用。从机制上来说，我们揭示了Src和PLC激酶信号传导的多效性细胞效应——细胞周期蛋白依赖性激酶1信号传导导致高尔基体特异性抗布雷菲德菌素A鸟嘌呤核苷酸交换因子1（GBF1）磷酸化以及经黄夹次苷处理后高尔基体囊泡化。GBF1磷酸化的后果是导致GBF1缺失，从而通过抑制病毒工厂的形成激活下游抗病毒信号传导。进一步研究表明，ERK1/2途径的信号传导——细胞周期蛋白依赖性激酶1激活导致GBF1在苏氨酸1337（T1337）处磷酸化。我们还表明，在经黄夹次苷处理的小鼠模型中，其保护率达100%，病毒滴度显著降低，且血清中无明显细胞毒性。这些发现凸显了解析广谱抗病毒治疗机制的重要性，并为在尚无疫苗可用的过渡阶段将黄夹次苷作为针对正链RNA病毒介导疾病的宿主导向型抗病毒药物的考虑铺平了道路。
