ABSTRACT

How severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infections engage cellular host pathways and innate immunity in infected cells remains largely elusive. We performed an integrative proteo-transcriptomics analysis in SARS-CoV-2 infected Huh7 cells to map the cellular response to the invading virus over time. We identified four pathways, ErbB, HIF-1, mTOR and TNF signaling, among others that were markedly modulated during the course of the SARS-CoV-2 infection in vitro. Western blot validation of the downstream effector molecules of these pathways revealed a dose-dependent activation of Akt, mTOR, S6K1 and 4E-BP1 at 24 hours post infection (hpi). However, we found a significant inhibition of HIF-1α through 24hpi and 48hpi of the infection, suggesting a crosstalk between the SARS-CoV-2 and the Akt/mTOR/HIF-1 signaling pathways. Inhibition of the mTOR signaling pathway using Akt inhibitor MK-2206 showed a significant reduction in virus production. Further investigations are required to better understand the molecular sequelae in order to guide potential therapy in the management of severe coronavirus disease 2019 (COVID-19) patients.

摘要

严重的急性呼吸系统综合症冠状病毒2（SARS-CoV-2）感染如何与细胞宿主途径和被感染细胞内的先天免疫力相互作用仍很遥远。我们在SARS-CoV-2感染的Huh7细胞中进行了综合蛋白转录组学分析，以绘制随时间推移对入侵病毒的细胞反应。我们确定了四种途径，ErbB，HIF-1，mTOR和TNF信号传导，以及在SARS-CoV-2体外感染过程中被显着调节的其他途径。这些途径的下游效应分子的蛋白质印迹验证表明，感染后24小时（hpi），Akt，mTOR，S6K1和4E-BP1呈剂量依赖性激活。但是，我们发现通过感染的24hpi和48hpi对HIF-1α有明显的抑制作用，这表明SARS-CoV-2与Akt / mTOR / HIF-1信号通路之间存在串扰。使用Akt抑制剂MK-2206抑制mTOR信号通路显示病毒产生显着减少。需要进一步的研究以更好地了解分子后遗症，以指导2019年严重冠状病毒病（COVID-19）患者管理中的潜在治疗方法。