Evidence of SARS-CoV-2 Transcriptional Activity in Cardiomyocytes of COVID-19 Patients without Clinical Signs of Cardiac Involvement

  1. Gaetano Pietro Bulfamante
  2. Gianluca Lorenzo Perrucci
  3. Monica Falleni
  4. Elena Sommariva
  5. Delfina Tosi
  6. Carla Martinelli
  7. Paola Songia
  8. Paolo Poggio
  9. Stefano Carugo
  10. Giulio Pompilio

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Abstract

Aims: A considerable proportion of patients affected by coronavirus respiratory disease (COVID-19) develop cardiac injury. The viral impact in cardiomyocytes deserves, however, further investigations, especially in asymptomatic patients. Methods: We investigated for SARS-CoV-2 presence and activity in heart tissues of six consecutive COVID-19 patients deceased from respiratory failure showing no signs of cardiac involvement and with no history of heart disease. Cardiac autopsy samples were collected within 2 h after death, and then analysed by digital PCR, Western blot, immunohistochemistry, immunofluorescence, RNAScope, and transmission electron microscopy assays. Results: The presence of SARS-CoV-2 into cardiomyocytes was invariably detected in all assays. A variable pattern of cardiomyocyte injury was observed, spanning from absence of cell death and subcellular alterations hallmarks, to intracellular oedema and sarcomere ruptures. In addition, we found active viral transcription in cardiomyocytes, by detecting both sense and antisense SARS-CoV-2 spike RNA. Conclusions: In this autopsy analysis of patients with no clinical signs of cardiac involvement, the presence of SARS-CoV-2 in cardiomyocytes has been detected, determining variable patterns of intracellular damage. These findings suggest the need for cardiologic surveillance in surviving COVID-19 patients not displaying a cardiac phenotype.

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  1. ScreenIT

    SciScore for 10.1101/2020.08.24.20170175: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablesamples: Autopsies of 6 consecutive non selected patients involved in this study, 5 males and 1 female, aged 54-69 years (average age: 59.5 ± 5.9 years) with a diagnosis of COVID-19, confirmed by CT analysis (Suppl. Fig. 1) and polymerase chain reaction (PCR), were performed in a BSL3 equipped autopsy room at the Unit of Pathology of “ASST Santi Paolo e Carlo” of Milan.

    Table 2: Resources

    Antibodies
    SentencesResources
    Primary antibodies adopted for western blot analysis were specific for ACE2 (R&D System, Minneapolis, MN, USA), SARS-CoV-2 nucleoprotein (Sino Biological, Düsseldorfer, Germany), SARS-CoV-2 spike protein (GeneTex, Irvine, CA, USA), and GAPDH (Santa Cruz Biotechnology, Dallas, TX, USA).
    ACE2
    suggested: None
    SARS-CoV-2 spike protein
    suggested: None
    GAPDH
    suggested: None
    Primary antibodies adopted for immunohistochemistry and immunofluorescence were specific for SARS-CoV-2 nucleoprotein (Sino Biological, Düsseldorfer, Germany), SARS-CoV-2 spike protein (GeneTex, Irvine, CA, USA), and α-SARC (AbCam, Cambridge, UK).
    α-SARC
    suggested: None
    Software and Algorithms
    SentencesResources
    Data availability: The raw data are available in Zenodo, at https://dx.doi.org/10.5281/zenodo.3974891.
    Zenodo
    suggested: (ZENODO, RRID:SCR_004129)
    Images were acquired with the ChemiDoc system (Bio-Rad Laboratories, Hercules, CA, USA).
    Bio-Rad Laboratories
    suggested: (Bio-Rad Laboratories, RRID:SCR_008426)
    The histochemistry with phosphotungstic acid haematoxylin (PTAH) was performed according to manufacturer’s protocol (BioOptica, Milan, Italy).
    BioOptica
    suggested: None

    Results from OddPub: Thank you for sharing your data.

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    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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  2. Version published to 10.3390/biomedicines8120626
  3. Version published to 10.1101/2020.08.24.20170175v1 on medRxiv