SARS-CoV2 infection impairs the metabolism and redox function of cellular glutathione
Desirée Bartolini,
Anna Maria Stabile,
Sabrina Bastianelli,
Daniela Giustarini,
Sara Pierucci,
Chiara Busti,
Carmine Vacca,
Anna Gidari,
Daniela Francisci,
Roberto Castronari,
Antonella Mencacci,
Manlio Di Cristina,
Riccardo Focaia,
Samuele Sabbatini,
Mario Rende,
Antimo Gioiello,
Gabriele Cruciani,
Ranieri Rossi,
Francesco Galli
Affiliations
Desirée Bartolini
Department of Pharmaceutical Sciences, University of Perugia, Nutrition and Clinical Biochemistry Lab, Via Del Giochetto, Monteluce, Perugia, Italy; Department of Medicine and Surgery, Section of Human, Clinical and Forensic Anatomy, School of Medicine, University of Perugia, P.le Lucio Severi, 1, Sant’Andrea Delle Fratte, 06132, Perugia, Italy
Anna Maria Stabile
Department of Medicine and Surgery, Section of Human, Clinical and Forensic Anatomy, School of Medicine, University of Perugia, P.le Lucio Severi, 1, Sant’Andrea Delle Fratte, 06132, Perugia, Italy
Sabrina Bastianelli
Department of Medicine and Surgery, Clinic of Infectious Diseases, University of Perugia, Perugia, Italy
Daniela Giustarini
Department of Biotechnology, Chemistry and Pharmacy, University of Siena, Via A. Moro 2, I-53100, Siena, Italy
Sara Pierucci
Department of Medicine and Surgery, Clinic of Infectious Diseases, University of Perugia, Perugia, Italy
Chiara Busti
Department of Medicine and Surgery, Clinic of Infectious Diseases, University of Perugia, Perugia, Italy
Carmine Vacca
Department of Chemistry, Biology and Biotechnology, University of Perugia, Via Elce di Sotto 8, 06123, Perugia, Italy
Anna Gidari
Department of Medicine and Surgery, Clinic of Infectious Diseases, University of Perugia, Perugia, Italy
Daniela Francisci
Department of Medicine and Surgery, Clinic of Infectious Diseases, University of Perugia, Perugia, Italy
Roberto Castronari
Department of Medicine and Surgery, Microbiology Unit, University of Perugia, 06123, Perugia, Italy
Antonella Mencacci
Department of Medicine and Surgery, Microbiology Unit, University of Perugia, 06123, Perugia, Italy
Manlio Di Cristina
Department of Chemistry, Biology and Biotechnology, University of Perugia, Via Elce di Sotto 8, 06123, Perugia, Italy
Riccardo Focaia
Department of Chemistry, Biology and Biotechnology, University of Perugia, Via Elce di Sotto 8, 06123, Perugia, Italy
Samuele Sabbatini
Department of Medicine and Surgery, Medical Microbiology Section, University of Perugia, 06129, Perugia, Italy
Mario Rende
Department of Medicine and Surgery, Section of Human, Clinical and Forensic Anatomy, School of Medicine, University of Perugia, P.le Lucio Severi, 1, Sant’Andrea Delle Fratte, 06132, Perugia, Italy
Antimo Gioiello
Department of Pharmaceutical Sciences, University of Perugia, Nutrition and Clinical Biochemistry Lab, Via Del Giochetto, Monteluce, Perugia, Italy
Gabriele Cruciani
Department of Chemistry, Biology and Biotechnology, University of Perugia, Via Elce di Sotto 8, 06123, Perugia, Italy
Ranieri Rossi
Department of Biotechnology, Chemistry and Pharmacy, University of Siena, Via A. Moro 2, I-53100, Siena, Italy
Francesco Galli
Department of Pharmaceutical Sciences, University of Perugia, Nutrition and Clinical Biochemistry Lab, Via Del Giochetto, Monteluce, Perugia, Italy; Corresponding author. Applied Biochemistry and Nutrition Lab, Dept. of Pharmaceutical Sciences, University of Perugia.
Viral infections sustain their replication cycle promoting a pro-oxidant environment in the host cell. In this context, specific alterations of the levels and homeostatic function of the tripeptide glutathione have been reported to play a causal role in the pro-oxidant and cytopathic effects (CPE) of the virus. In this study, these aspects were investigated for the first time in SARS-CoV2-infected Vero E6 cells, a reliable and well-characterized in vitro model of this infection. SARS-CoV2 markedly decreased the levels of cellular thiols, essentially lowering the reduced form of glutathione (GSH). Such an important defect occurred early in the CPE process (in the first 24 hpi). Thiol analysis in N-acetyl-Cys (NAC)-treated cells and membrane transporter expression data demonstrated that both a lowered uptake of the GSH biosynthesis precursor Cys and an increased efflux of cellular thiols, could play a role in this context. Increased levels of oxidized glutathione (GSSG) and protein glutathionylation were also observed along with upregulation of the ER stress marker PERK. The antiviral drugs Remdesivir (Rem) and Nelfinavir (Nel) influenced these changes at different levels, essentially confirming the importance or blocking viral replication to prevent GSH depletion in the host cell. Accordingly, Nel, the most potent antiviral in our in vitro study, produced a timely activation of Nrf2 transcription factor and a GSH enhancing response that synergized with NAC to restore GSH levels in the infected cells. Despite poor in vitro antiviral potency and GSH enhancing function, Rem treatment was found to prevent the SARS-CoV2-induced glutathionylation of cellular proteins. In conclusion, SARS-CoV2 infection impairs the metabolism of cellular glutathione. NAC and the antiviral Nel can prevent such defect in vitro.
