PDF(Pubmed)
Abstract：
Pyroptosis, a pro-inflammatory programmed cell death, has been implicated in the pathogenesis of coronavirus disease 2019 and other viral diseases. Gasdermin family proteins (GSDMs), including GSDMD and GSDME, are key regulators of pyroptotic cell death. However, the mechanisms by which virus infection modulates pyroptosis remain unclear. Here, we employed a mCherry-GSDMD fluorescent reporter assay to screen for viral proteins that impede the localization and function of GSDMD in living cells. Our data indicated that the main protease NSP5 of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) blocked GSDMD-mediated pyroptosis via cleaving residues Q29 and Q193 of GSDMD. While another SARS-CoV-2 protease, NSP3, cleaved GSDME at residue G370 but activated GSDME-mediated pyroptosis. Interestingly, respiratory enterovirus EV-D68-encoded proteases 3C and 2A also exhibit similar differential regulation on the functions of GSDMs by inactivating GSDMD but initiating GSDME-mediated pyroptosis. EV-D68 infection exerted oncolytic effects on human cancer cells by inducing pyroptotic cell death. Our findings provide insights into how respiratory viruses manipulate host cell pyroptosis and suggest potential targets for antiviral therapy as well as cancer treatment.IMPORTANCEPyroptosis plays a crucial role in the pathogenesis of coronavirus disease 2019, and comprehending its function may facilitate the development of novel therapeutic strategies. This study aims to explore how viral-encoded proteases modulate pyroptosis. We investigated the impact of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and respiratory enterovirus D68 (EV-D68) proteases on host cell pyroptosis. We found that SARS-CoV-2-encoded proteases NSP5 and NSP3 inactivate gasdermin D (GSDMD) but initiate gasdermin E (GSDME)-mediated pyroptosis, respectively. We also discovered that another respiratory virus EV-D68 encodes two distinct proteases 2A and 3C that selectively trigger GSDME-mediated pyroptosis while suppressing the function of GSDMD. Based on these findings, we further noted that EV-D68 infection triggers pyroptosis and produces oncolytic effects in human carcinoma cells. Our study provides new insights into the molecular mechanisms underlying virus-modulated pyroptosis and identifies potential targets for the development of antiviral and cancer therapeutics.
摘要：
焦亡,促炎的程序性细胞死亡，与2019年冠状病毒病和其他病毒性疾病的发病机理有关。Gasdermin家族蛋白(GSDMs),包括GSDMD和GSDME，是焦转细胞死亡的关键调节因子。然而,病毒感染调节焦亡的机制尚不清楚.这里,我们采用mCherry-GSDMD荧光报告基因检测方法来筛选阻碍GSDMD在活细胞中定位和功能的病毒蛋白.我们的数据表明，严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的主要蛋白酶NSP5通过切割GSDMD的残基Q29和Q193阻断了GSDMD介导的焦亡。而另一种SARS-CoV-2蛋白酶，NSP3在残基G370处切割GSDME,但激活GSDME介导的焦亡。有趣的是,呼吸道肠道病毒EV-D68编码的蛋白酶3C和2A也通过灭活GSDMD但启动GSDME介导的焦亡而对GSDM的功能表现出相似的差异调节。EV-D68感染通过诱导细胞凋亡对人癌细胞产生溶瘤作用。我们的发现提供了有关呼吸道病毒如何操纵宿主细胞焦亡的见解，并为抗病毒治疗和癌症治疗提供了潜在的靶标。IMPORTANCEPyroptosis在2019冠状病毒病的发病机理中起着至关重要的作用，理解其功能可能有助于开发新的治疗策略。本研究旨在探讨病毒编码的蛋白酶如何调节焦亡。我们研究了严重急性呼吸综合征冠状病毒2（SARS-CoV-2）和呼吸道肠道病毒D68（EV-D68）蛋白酶对宿主细胞焦亡的影响。我们发现SARS-CoV-2编码的蛋白酶NSP5和NSP3使gasderminD（GSDMD）失活，但引发gasderminE（GSDME）介导的焦亡，分别。我们还发现另一种呼吸道病毒EV-D68编码两种不同的蛋白酶2A和3C，它们选择性触发GSDME介导的焦亡，同时抑制GSDMD的功能。基于这些发现,我们进一步注意到,EV-D68感染引发人癌细胞的焦亡并产生溶瘤作用.我们的研究为病毒调节的焦亡的分子机制提供了新的见解，并确定了抗病毒和癌症疗法发展的潜在靶标。
