Pericyte-specific vascular expression of SARS-CoV-2 receptor ACE2 – implications for microvascular inflammation and hypercoagulopathy in COVID-19

  1. Liqun He
  2. Maarja Andaloussi Mäe
  3. Lars Muhl
  4. Ying Sun
  5. Riikka Pietilä
  6. Khayrun Nahar
  7. Elisa Vázquez Liébanas
  8. Malin Jonsson Fagerlund
  9. Anders Oldner
  10. Jianping Liu
  11. Guillem Genové
  12. Lei Zhang
  13. Yuan Xie
  14. Stefanos Leptidis
  15. Giuseppe Mocci
  16. Simon Stritt
  17. Ahmed Osman
  18. Andrey Anisimov
  19. Karthik Amudhala Hemanthakumar
  20. Markus Räsänen
  21. Olivier Mirabeau
  22. Emil Hansson
  23. Johan Björkegren
  24. Michael Vanlandewijck
  25. Klas Blomgren
  26. Taija Mäkinen
  27. Xiao-Rong Peng
  28. Thomas D. Arnold
  29. Kari Alitalo
  30. Lars I Eriksson
  31. Urban Lendahl
  32. Christer Betsholtz

Reviewed by

Read the full article

Listed in

Log in to save this article

Abstract

Accumulating clinical observations implicate vascular inflammation as an underlying cause of coagulopathy in severely ill COVID-19 patients and it was recently suggested that SARS-CoV-2 virus particles infect endothelial cells. Here, we show that endothelial cells do not express angiotensin-converting enzyme-2 (ACE2), the SARS-CoV-2 receptor. Instead, pericytes and microvascular smooth muscle cells express ACE2 in an organotypic manner. Pericyte deficiency leads to increased endothelial expression and release of Von Willebrand factor and intravascular platelet and fibrin aggregation, suggesting that pericytes limit endothelial pro-thrombotic responses. That pericytes and not endothelial cells express ACE2 may provide important clues to the pathology of COVID-19, as pericytes are normally shielded behind an endothelial barrier and may get infected only when this barrier is compromised by COVID-19 risk factors.

Article activity feed

  1. Version published to 10.1101/2020.05.11.088500v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2020.05.11.088500: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementAnimal protocols were approved by either the Uppsala Ethical Committee on Animal Research ( permit numbers C224/12 , C115/15 , C111515/16) , or by the Stockholm/Linköping Ethical Committee on Animal Research ( permit ID 729) .RandomizationExpression of selected known pericyte specific markers in 23 Ace2-positive endothelial cells ( top ) and 23 randomly selected Ace2-negative endothelial cells ( bottom) .Blindingnot detected.Power Analysisnot detected.Sex as a biological variableThe ICU patients had a median age of 63 years (range 4073), the majority being male (90%) and having one or more risk factors prior to admission (85%).Cell Line AuthenticationFocusing on the brain cortex , where vascular arterio-venous identities can readily be identified through morphology and marker expression , we found ACE2-positive cells fitting the expected location and marker expression of pericytes and microvascular VSMCs ( Fig 1D and Suppl Fig 2) .

    Table 2: Resources

    Antibodies
    SentencesResources
    We also assessed a human glioblastoma drop-seq 10x scRNAseq dataset consisting of 69,125 endothelial and immune cells sorted using anti-CD31 antibodies ( Zhang et al , unpublished) .
    anti-CD31
    suggested: None
    IF staining of heart tissue from Pdgfrb-GFP reporter mice using antibodies against ACE2 and CD31 .
    antibodies against ACE2
    suggested: (GeneTex Cat# GTX123624, AB_11177358)
    IF staining of mouse cortex vasculature in Pdgfbret/+ and Pdgfbret/ret mice with antibodies against ( B ) VWF , PDGFRB and ACTA2 , and ( C ) PODXL , PDGFRB and VWF) . a , arterioles . v , venules .
    ACTA2
    suggested: None
          <div style="margin-bottom:8px">
        <div><b>VWF</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Software and Algorithms</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">7 Institute of Genomics and Multiscale Biology, Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, NY, US 8 Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia 9 Clinical Gene Networks AB, Stockholm, Sweden Cardiovascular, Renal and Metabolism (CVRM), IMED Biotech Unit, AstraZeneca BioPharmaceuticals R&D, Gothenburg, Sweden 11 Department of Pediatric oncology, Karolinska University Hospital, Stockholm, Sweden 12 Department of Pediatrics, University of California San Francisco, USA 13 Department of Cell and Molecular Biology, Karolinska Institutet, Sweden 14 Department of Medicine Huddinge, Karolinska Institutet, Sweden Abstract Accumulating clinical observations suggest pathogenesis beyond viral pneumonia and its secondary consequences in COVID-19 patients.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>AstraZeneca BioPharmaceuticals</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">For integration of different datasets , the integration workflow “Reciprocal PCA” in the Seurat package was implemented , which integrated overall datasets using the mutual nearest neighbor ( MNN ) cell pairs that shared a common set of molecular features in their PCA spaces .</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Seurat</b></div>
        <div>suggested: (SEURAT, <a href="https://scicrunch.org/resources/Any/search?q=SCR_007322">SCR_007322</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Their expression profiles in Ace2-positive and Ace2-negative cells were compared in a random selection of equal numbers of cells , and the heat map results were visualized with pheatmap package ( version 1.0.12 ) in R software.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>pheatmap</b></div>
        <div>suggested: (pheatmap, <a href="https://scicrunch.org/resources/Any/search?q=SCR_016418">SCR_016418</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">All confocal images are represented as maximum intensity projections and were adjusted for brightness and contrast using Fiji v1.52p and Adobe Photoshop CC 2019</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Fiji</b></div>
        <div>suggested: (Fiji, <a href="https://scicrunch.org/resources/Any/search?q=SCR_002285">SCR_002285</a>)</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>Adobe Photoshop</b></div>
        <div>suggested: (Adobe Photoshop, <a href="https://scicrunch.org/resources/Any/search?q=SCR_014199">SCR_014199</a>)</div>
      </div>
    </td></tr></table>

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

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.

  3. Version published to 10.1101/2020.05.11.088500v1 on bioRxiv