TMPRSS2-mediated SARS- CoV-2 uptake boosts innate immune activation, enhances cytopathology, and drives convergent virus evolution

• 成果类型: Article
• 署名作者: Qu, Bingqian; Miskey, Csaba; Goemer, Andre; Kleinert, Robin D. V.; Ibanez, Sara Calvo; Eberle, Regina; Ebenig, Aileen; Postmus, Dylan; Nocke, Maximilian K.; Herrmann, Maike; Itotia, Tabitha K.; Herrmann, Simon T.; Heinen, Natalie; Hoeck, Sebastian; Hastert, Florian D.; von Rhein, Christine; Schuermann, Christoph; Li, Xue; van Zandbergen, Ger; Widera, Marek; Ciesek, Sandra; Schnierle, Barbara S.; Tarr, Alexander W.; Steinmann, Eike; Goffinet, Christine; Pfaender, Stephanie; Locker, Jacomina Krijnse; Muehlebach, Michael D.; Todt, Daniel; Brown, Richard J. P.
• 署名单位: Paul Ehrlich Institute; Paul Ehrlich Institute; Ruhr University Bochum; Paul Ehrlich Institute; Liverpool School of Tropical Medicine; Free University of Berlin; Humboldt University of Berlin; Charite Universitatsmedizin Berlin; Paul Ehrlich Institute; University of Hamburg; University Medical Center Hamburg-Eppendorf; Ruprecht Karls University Heidelberg; Paul Ehrlich Institute; Johannes Gutenberg University of Mainz; Johannes Gutenberg University of Mainz; Goethe University Frankfurt; Goethe University Frankfurt Hospital; German Center for Infection Research; University of Nottingham; University of Nottingham; University of Nottingham; University of Lubeck; German Center for Infection Research
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-11158
• DOI: 10.1073/pnas.2407437121
• 发表日期: 2024-06-04
• 关键词: cell entry; tmprss2; sars-cov-2

摘要

The accessory protease transmembrane protease serine 2 (TMPRSS2) enhances severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) uptake into ACE2-expressing cells, although how increased entry impacts downstream viral and host processes remains unclear. To investigate this in more detail, we performed infection assays in engineered cells promoting ACE2-mediated entry with and without TMPRSS2 coexpression. Electron microscopy and inhibitor experiments indicated TMPRSS2-mediated cell entry was associated with increased virion internalization into endosomes, and partially dependent upon clathrin-mediated endocytosis. TMPRSS2 increased panvariant uptake efficiency and enhanced early rates of virus replication, transcription, and secretion, with variant-specific profiles observed. On the host side, transcriptional profiling confirmed the magnitude of infection-induced antiviral and proinflammatory responses were linked to uptake efficiency, with TMPRSS2-assisted entry boosting early antiviral responses. In addition, TMPRSS2-enhanced infections increased rates of cytopathology, apoptosis, and necrosis and modulated virus secretion kinetics in a variant-specific manner. On the virus side, convergent signatures of cell-uptake-dependent innate immune induction were recorded in viral genomes, manifesting as switches in dominant coupled Nsp3 residues whose frequencies were correlated to the magnitude of the cellular response to infection. Experimentally, we demonstrated that selected Nsp3 mutations conferred enhanced interferon antagonism. More broadly, we show that TMPRSS2 orthologues from evolutionarily diverse mammals facilitate panvariant enhancement of cell uptake. In summary, our study uncovers previously unreported associations, linking cell entry efficiency to innate immune activation kinetics, cell death rates, virus secretion dynamics, and convergent selection of viral mutations. These data expand our understanding of TMPRSS2's role in the SARS-CoV-2 life cycle and confirm its broader significance in zoonotic reservoirs and animal models.