Cytosolic proliferating cell nuclear antigen (PCNA) orchestrates neutrophil hyperactivation in COVID-19
Abstract
Neutrophils are central mediators of the hyperinflammatory response in severe SARS-CoV-2 infection. We report elevated cytosolic levels of proliferating cell nuclear antigen (PCNA) in neutrophils from patients with severe and critical COVID-19, correlating with enhanced NADPH oxidase–dependent reactive oxygen species (ROS) generation and neutrophil extracellular trap (NET) formation. Using T2AA, a small-molecule inhibitor of the PCNA scaffold, we demonstrate potent suppression of NADPH oxidase activation and NET release, particularly in response to SARS-CoV-2 RNA. Mechanistically, we identify a previously unrecognized interaction between PCNA and the heterodimeric S100A8/S100A9 (calprotectin), predominantly enriched in CD16 high CD62L low neutrophils expanded during COVID-19. PCNA binds the dimeric S100A8/S100A9 complex mediated via S100A8 subunit with micromolar affinity, and this interaction is abrogated by tetramerization, suggesting regulation by intracellular calcium. Disruption of this complex by T2AA inhibited ROS production in an S100A8/S100A9-dependent manner, implicating calprotectin as a functional regulator of neutrophil activation. In a betacoronavirus mouse model, T2AA treatment attenuated lung inflammation, reduced NET and calprotectin levels, and shifted pulmonary neutrophils away from hyperactivated and immunosuppressive phenotypes, consistent with immune reprogramming toward resolution. These findings establish cytosolic PCNA as a central scaffold in neutrophil hyperactivation during COVID-19 and highlight its pharmacological disruption as a promising host-directed strategy to limit inflammation and prevent organ damage.
Article Details
Journal Info
Proceedings of the National Academy of Sciences
National Academy of Sciences
Authors (46)
Rodrigo de Oliveira Formiga
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Lucie Pesenti
Université Paris Cité, CNRS UMR 8104, Inserm U1016
François Chable de la Héronnière
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Maha Zohra Ladjemi
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Darko Stojkov
Shida Yousefi
Institute of Pharmacology, University of Bern
Philippe Frachet
Université Grenoble Alpes, CNRS, Institute of Structural Biology (IBS) - UMR 5075
Lisa Krafft
Institute of Immunology, University of Münster
Laura Tiberio
Department of Molecular and Translational Medicine, University of Brescia
Daniela Bosisio
Department of Molecular and Translational Medicine, University of Brescia
Muriel Andrieu
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Souganya Many
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Vaarany Karunanithy
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Karine Bailly
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Théo Dhôte
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Giovanni Saraceni-Tasso
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Manon Castel
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Christophe Rousseau
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Marick Rodrigues Starick
Department of Pharmacology, Federal University of Santa Catarina
Edroaldo Lummertz da Rocha
Laboratory of Systems Biology, Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina
Emilia Puig Lombardi
Bioinformatics Core Platform, INSERM Unit 1163, Imagine Institute, Université Paris Cité
Cicero José Luíz dos Ramos Almeida
Center for Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo
Anderson dos Santos Ramos
Center for Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo
Fernando Queiroz Cunha
Center for Research in Inflammatory Diseases, Ribeirão Preto Medical School, University of São Paulo
Jose Carlos Alves-Filho
Natália Ribeiro Cabacinha Nóbrega
Department of Morphology, Institute of Biological Sciences, Federal University of Minas Gerais
Matheus Rodrigues Gonçalves
Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais
Celso Martins Queiroz-Junior
Department of Morphology, Institute of Biological Sciences, Federal University of Minas Gerais
Viviane Lima Batista
Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais
Mauro Martins Teixeira
Department of Biochemistry and Immunology, Institute of Biological Sciences, Federal University of Minas Gerais
Vanessa Granger
Assistance Publique–Hôpitaux de Paris (AP-HP), Department of Immunology, Bichat Hospital
Sylvie Chollet-Martin
Assistance Publique–Hôpitaux de Paris (AP-HP), Department of Immunology, Bichat Hospital
Luc De Chaisemartin
Assistance Publique–Hôpitaux de Paris (AP-HP), Department of Immunology, Bichat Hospital
Luc Mouthon
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Anne Hosmalin
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Margarita Hurtado-Nedelec
Department of Immunology and Hematology, Functional Unit of Immune Dysfunctions, Hôpitaux Universitaires Paris Nord Val-de-Seine, Bichat Hospital
Clémence Martin
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Fernando Spiller
Department of Pharmacology, Federal University of Santa Catarina
Hans-Uwe Simon
Institute of Pharmacology, University of Bern
Nicolas Tamassia
Department of Medicine, Section of General Pathology, University of Verona
Marco Antonio Cassatella
Department of Medicine, Section of General Pathology, University of Verona
Frédéric Pène
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Thomas Vogl
Institute of Immunology, University of Münster
Pierre-Regis Burgel
Université Paris Cité, CNRS UMR 8104, Inserm U1016
Vivian Vasconcelos Costa
Department of Morphology, Institute of Biological Sciences, Federal University of Minas Gerais
Véronique Witko-Sarsat
Université Paris Cité, CNRS UMR 8104, Inserm U1016