Pulmonary infection by SARS-CoV-2 induces senescence accompanied by an inflammatory phenotype in severe COVID-19: possible implications for viral mutagenesis

Konstantinos Evangelou; Dimitris Veroutis; Koralia Paschalaki; Periklis G Foukas; Nefeli Lagopati; Marios Dimitriou; Angelos Papaspyropoulos; Bindu Konda; Orsalia Hazapis; Aikaterini Polyzou; Sophia Havaki; Athanassios Kotsinas; Christos Kittas; Athanasios G Tzioufas; Laurence de Leval; Demetris Vassilakos; Sotirios Tsiodras; Barry R Stripp; Argyris Papantonis; Giovanni Blandino; Ioannis Karakasiliotis; Peter J Barnes; Vassilis G Gorgoulis

doi:10.1183/13993003.02951-2021

Pulmonary infection by SARS-CoV-2 induces senescence accompanied by an inflammatory phenotype in severe COVID-19: possible implications for viral mutagenesis

Eur Respir J. 2022 Aug 18;60(2):2102951. doi: 10.1183/13993003.02951-2021. Print 2022 Aug.

Authors

Konstantinos Evangelou^{1

2}, Dimitris Veroutis^{1

3

2}, Koralia Paschalaki^{4

2}, Periklis G Foukas⁵, Nefeli Lagopati^{1

3}, Marios Dimitriou^{5

6}, Angelos Papaspyropoulos^{1

3}, Bindu Konda⁷, Orsalia Hazapis¹, Aikaterini Polyzou¹, Sophia Havaki¹, Athanassios Kotsinas¹, Christos Kittas^{1

8}, Athanasios G Tzioufas⁹, Laurence de Leval¹⁰, Demetris Vassilakos¹, Sotirios Tsiodras^{11

12}, Barry R Stripp⁷, Argyris Papantonis^{13

14}, Giovanni Blandino¹⁵, Ioannis Karakasiliotis⁶, Peter J Barnes^{4

16}, Vassilis G Gorgoulis^{17

3

8

18

19

16}

Affiliations

¹ Molecular Carcinogenesis Group, Dept of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
² Contributed equally.
³ Biomedical Research Foundation, Academy of Athens, Athens, Greece.
⁴ National Heart and Lung Institute, Imperial College London, London, UK.
⁵ 2nd Dept of Pathology, Attikon University Hospital, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
⁶ Laboratory of Biology, Dept of Medicine, Democritus University of Thrace, Alexandroupolis, Greece.
⁷ Lung and Regenerative Medicine Institutes, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
⁸ Faculty of Health and Medical Sciences, University of Surrey, Guildford, UK.
⁹ Dept of Pathophysiology, Medical School, National and Kapodistrian University of Athens, Athens, Greece.
¹⁰ Institute of Pathology, Lausanne University Hospital, Lausanne, Switzerland.
¹¹ 4th Dept of Internal Medicine, Attikon University Hospital, University of Athens Medical School, Athens, Greece.
¹² Hellenic Centre for Disease Control and Prevention, Athens, Greece.
¹³ Translational Epigenetics Group, Institute of Pathology, University Medical Center Göttingen, Göttingen, Germany.
¹⁴ Center for Molecular Medicine, University of Cologne, Cologne, Germany.
¹⁵ Oncogenomic and Epigenetic Unit, IRCCS, Regina Elena National Cancer Institute, Rome, Italy.
¹⁶ P.J. Barnes and V.G. Gorgoulis contributed equally to this article as lead authors and supervised the work.
¹⁷ Molecular Carcinogenesis Group, Dept of Histology and Embryology, Medical School, National and Kapodistrian University of Athens, Athens, Greece [email protected].
¹⁸ Faculty Institute for Cancer Sciences, Manchester Academic Health Sciences Centre, University of Manchester, Manchester, UK.
¹⁹ Center for New Biotechnologies and Precision Medicine, Medical School, National and Kapodistrian University of Athens, Athens, Greece.

Abstract

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection of the respiratory system can progress to a multisystemic disease with aberrant inflammatory response. Cellular senescence promotes chronic inflammation, named senescence-associated secretory phenotype (SASP). We investigated whether coronavirus disease 2019 (COVID-19) is associated with cellular senescence and SASP.

Methods: Autopsy lung tissue samples from 11 COVID-19 patients and 43 age-matched non-COVID-19 controls with similar comorbidities were analysed by immunohistochemistry for SARS-CoV-2, markers of senescence and key SASP cytokines. Virally induced senescence was functionally recapitulated in vitro, by infecting epithelial Vero-E6 cells and a three-dimensional alveosphere system of alveolar type 2 (AT2) cells with SARS-CoV-2 strains isolated from COVID-19 patients.

Results: SARS-CoV-2 was detected by immunocytochemistry and electron microscopy predominantly in AT2 cells. Infected AT2 cells expressed angiotensin-converting enzyme 2 and exhibited increased senescence (p16^INK4A and SenTraGor positivity) and interleukin (IL)-1β and IL-6 expression. In vitro, infection of Vero-E6 cells with SARS-CoV-2 induced senescence (SenTraGor), DNA damage (γ-H2AX) and increased cytokine (IL-1β, IL-6, CXCL8) and apolipoprotein B mRNA-editing (APOBEC) enzyme expression. Next-generation sequencing analysis of progenies obtained from infected/senescent Vero-E6 cells demonstrated APOBEC-mediated SARS-CoV-2 mutations. Dissemination of the SARS-CoV-2-infection and senescence was confirmed in extrapulmonary sites (kidney and liver) of a COVID-19 patient.

Conclusions: We demonstrate that in severe COVID-19, AT2 cells infected by SARS-CoV-2 exhibit senescence and a proinflammatory phenotype. In vitro, SARS-CoV-2 infection induces senescence and inflammation. Importantly, infected senescent cells may act as a source of SARS-CoV-2 mutagenesis mediated by APOBEC enzymes. Therefore, SARS-CoV-2-induced senescence may be an important molecular mechanism of severe COVID-19, disease persistence and mutagenesis.

MeSH terms

COVID-19*
Cellular Senescence
Cytokines / metabolism
Humans
Inflammation
Interleukin-6
Lung / metabolism
Mutagenesis
Phenotype
SARS-CoV-2*

Substances

Cytokines
Interleukin-6