­­­­­­­A type I IFN, prothrombotic hyperinflammatory neutrophil signature is distinct for COVID-19 ARDS­­­

  1. Leila Reyes
  2. Manuel A. Sanchez-Garcia
  3. Tyler Morrison
  4. Andy J. M. Howden
  5. Emily R. Watts
  6. Simone Arienti
  7. Pranvera Sadiku
  8. Patricia Coelho
  9. Ananda S. Mirchandani
  10. Ailiang Zhang
  11. David Hope
  12. Sarah K. Clark
  13. Jo Singleton
  14. Shonna Johnston
  15. Robert Grecian
  16. Azin Poon
  17. Sarah McNamara
  18. Isla Harper
  19. Max Head Fourman
  20. Alejandro J. Brenes
  21. Shalini Pathak
  22. Amy Lloyd
  23. Giovanny Rodriguez Blanco
  24. Alex von Kriegsheim
  25. Bart Ghesquiere
  26. Wesley Vermaelen
  27. Camila T. Cologna
  28. Kevin Dhaliwal
  29. Nik Hirani
  30. David H. Dockrell
  31. Moira K. B. Whyte
  32. David Griffith
  33. Doreen A. Cantrell
  34. Sarah R. Walmsley

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Abstract

Background: Acute respiratory distress syndrome (ARDS) is a severe critical condition with a high mortality that is currently in focus given that it is associated with mortality caused by coronavirus disease 2019 (COVID-19). Neutrophils play a key role in the lung injury characteristic of non-COVID-19 ARDS and there is also accumulating evidence of neutrophil mediated lung injury in patients who succumb to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

Methods: We undertook a functional proteomic and metabolomic survey of circulating neutrophil populations, comparing patients with COVID-19 ARDS and non-COVID-19 ARDS to understand the molecular basis of neutrophil dysregulation.

Results: Expansion of the circulating neutrophil compartment and the presence of activated low and normal density mature and immature neutrophil populations occurs in ARDS, irrespective of cause. Release of neutrophil granule proteins, neutrophil activation of the clotting cascade and upregulation of the Mac-1 platelet binding complex with formation of neutrophil platelet aggregates is exaggerated in COVID-19 ARDS. Importantly, activation of components of the neutrophil type I interferon responses is seen in ARDS following infection with SARS-CoV-2, with associated rewiring of neutrophil metabolism, and the upregulation of antigen processing and presentation. Whilst dexamethasone treatment constricts the immature low density neutrophil population, it does not impact upon prothrombotic hyperinflammatory neutrophil signatures.

Conclusions: Given the crucial role of neutrophils in ARDS and the evidence of a disordered myeloid response observed in COVID-19 patients, this work maps the molecular basis for neutrophil reprogramming in the distinct clinical entities of COVID-19 and non-COVID-19 ARDS.

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  1. Version published to 10.12688/wellcomeopenres.16584.2
  2. ScreenIT

    SciScore for 10.1101/2020.09.15.20195305: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board StatementConsent: Healthy donor and patient recruitment: Human peripheral venous blood was taken from healthy volunteers with written informed consent obtained from all participants prior to sample collection as approved by the University of Edinburgh Centre for Inflammation Research Blood Resource Management Committee (AMREC 15-HV-013).
    IRB: The collection of peripheral venous blood from patients diagnosed with COVID-19 and/or presenting with ARDS was approved by Scotland A Research Ethics Committee.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Cells were pelleted and blocked with Fc Receptor Blocking Solution followed by staining with anti-CD41 antibody (Biolegend) and counterstaining with propidium iodide (Biolegend) according to manufacturer’s guidelines.
    anti-CD41
    suggested: None
    Software and Algorithms
    SentencesResources
    Patient recruitment took place from April 2020 through January 2021 from The Royal Infirmary of Edinburgh, Scotland, UK through the ARDS Neut (20/SS/0002) and CASCADE (20/SS/0052) Study, with informed consent obtained by proxy.
    CASCADE
    suggested: (Cascade, RRID:SCR_005861)
    Compensation was performed using BD FACSDiva™ software version 8.0 and data analysed in FlowJo version 10.2.
    BD FACSDiva™
    suggested: (BD FACSDiva Software, RRID:SCR_001456)
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    PepMap nanoViper C18 column, 5 μm, 100 Å, Thermo Scientific) equilibrated in 0.1% trifluoroacetic acid (TFA).
    PepMap
    suggested: (BioWorks, RRID:SCR_014594)
    Fold changes and P-values were calculated in R utilising the bioconductor package LIMMA version 3.7(28).
    bioconductor
    suggested: (Bioconductor, RRID:SCR_006442)
    LIMMA
    suggested: (LIMMA, RRID:SCR_010943)
    Raw mass spectrometry data files and Spectronaut analysis files have been deposited to the ProteomeXchange (30) Consortium via the PRIDE (31) partner repository.
    ProteomeXchange
    suggested: (ProteomeXchange, RRID:SCR_004055)
    PRIDE
    suggested: (Pride-asap, RRID:SCR_012052)
    FACS of NDN and LDN from PMN and PBMC layers respectively were performed using BD FACSAria™ Fusion flow cytometer fitted with a 70 µm nozzle and running BD FACSDiva™ software version 8.0 (Beckton Dickinson).
    BD FACSAria™
    suggested: (BD FACSAria II Cell Sorter, RRID:SCR_018934)
    Fiji software was used to process the images (32).
    Fiji
    suggested: (Fiji, RRID:SCR_002285)
    Statistical analyses: Statistical tests were performed using Prism 9.00 software (GraphPad Software Inc).
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:
    A clear limitation of this study is the relatively small number of patients recruited. This is balanced against the detailed analysis we have been able to perform in these patient groups and the move towards functional dissection of neutrophil responses not previously captured in transcriptional data sets. Further understanding of the mechanisms which regulate aberrant neutrophil responses will likely be important in developing strategies to target the innate responses following infection with SARS-CoV-2 to enable an effective therapeutic arsenal for COVID-19 ARDS.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

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  3. Version published to 10.1101/2020.09.15.20195305v2 on medRxiv
  4. Version published to 10.1101/2020.09.15.20195305v1 on medRxiv