Gasdermin-D activation by SARS-CoV-2 triggers NET and mediate COVID-19 immunopathology
Camila M. Silva, Carlos W. Wanderley, Flávio P. Veras, Augusto V. Gonçalves, Mikhael Haruo Fernandes de Lima, Juliana E. Toller-Kawahisa, Giovanni Freitas Gomes, Daniele C. Nascimento, Valter Silva Monteiro, Isadora Marques Paiva, Cicero J. L. R. Almeida, Diego B. Caetité, Juliana Costa Silva, Maria Isabel Fernandes Lopes, Letícia Pastorelli Bonjorno, Marcela C Giannini, Natália B. do Amaral, Maíra Nilson Benatti, Rodrigo de Carvalho Santana, Luis Eduardo Alves Damasceno, Bruna Manuella Souza Silva, Ayda Henriques Schneider, Ícaro Maia Santos de Castro, Juan Carlo Santos e Silva, Amanda Pereira Vasconcelos, Tiago Tomazini Gonçalves, Sabrina Setembre Batah, Tamara Silva Rodrigues, Victor Ferreira Costa, Marjorie Cornejo Pontelli, Ronaldo B. Martins, Timna Varela Martins, Danillo Lucas Alves Espósito, Guilherme Cesar Martelossi Cebinelli, Benedito Antônio Lopes da Fonseca, Luiz Osório Leiria, Larissa D. Cunha, Eurico Arruda, Helder I. Nakaia, Alexandre Todorovic Fabro, Renê Donizeti Ribeiro de Oliveira, Dario S. Zamboni, Paulo Louzada‐Júnior, Thiago M. Cunha, José Carlos Farias Alves-Filho, Fernando Q. Cunha
Abstract
BACKGROUND: The release of neutrophil extracellular traps (NETs) is associated with inflammation, coagulopathy, and organ damage found in severe cases of COVID-19. However, the molecular mechanisms underlying the release of NETs in COVID-19 remain unclear. OBJECTIVES: We aim to investigate the role of the Gasdermin-D (GSDMD) pathway on NETs release and the development of organ damage during COVID-19. METHODS: We performed a single-cell transcriptome analysis in public data of bronchoalveolar lavage. Then, we enrolled 63 hospitalized patients with moderate and severe COVID-19. We analyze in blood and lung tissue samples the expression of GSDMD, presence of NETs, and signaling pathways upstreaming. Furthermore, we analyzed the treatment with disulfiram in a mouse model of SARS-CoV-2 infection. RESULTS: We found that the SARS-CoV-2 virus directly activates the pore-forming protein GSDMD that triggers NET production and organ damage in COVID-19. Single-cell transcriptome analysis revealed that the expression of GSDMD and inflammasome-related genes were increased in COVID-19 patients. High expression of active GSDMD associated with NETs structures was found in the lung tissue of COVID-19 patients. Furthermore, we showed that activation of GSDMD in neutrophils requires active caspase1/4 and live SARS-CoV-2, which infects neutrophils. In a mouse model of SARS-CoV-2 infection, the treatment with disulfiram inhibited NETs release and reduced organ damage. CONCLUSION: These results demonstrated that GSDMD-dependent NETosis plays a critical role in COVID-19 immunopathology and suggests GSDMD as a novel potential target for improving the COVID-19 therapeutic strategy.