Plasma mediators in patients with severe COVID-19 cause lung endothelial barrier failure

Laura Michalick¹, Sarah Weidenfeld¹, Benjamin Grimmer¹, Diana Fatykhova², Philip Daniel Solymosi¹, Felix Behrens^{1

3}, Melanie Dohmen^{2

4}, Markus C Brack², Sabrina Schulz¹, Elisa Thomasch¹, Szandor Simmons¹, Holger Müller-Redetzky², Norbert Suttorp^{2

5}, Florian Kurth², Jens Neudecker⁶, Mario Toennies⁷, Torsten T Bauer⁷, Stephan Eggeling⁸, Victor Max Corman^{9

10}, Andreas C Hocke^{2

5}, Martin Witzenrath^{2

5

11}, Stefan Hippenstiel^{2

5

11}, Wolfgang M Kuebler^{12

5

13

14

15

11}

Affiliations

¹ Institute of Physiology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
² Dept of Infectious Diseases and Respiratory Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
³ Berlin Institute of Health (BIH), Berlin, Germany.
⁴ Max Delbrück Center for Molecular Medicine, Berlin, Germany.
⁵ German Center for Lung Research (DZL), Germany.
⁶ Dept of General, Visceral, Vascular and Thoracic Surgery, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
⁷ Lungenklinik Heckeshorn, HELIOS Klinikum Emil von Behring, Berlin, Germany.
⁸ Vivantes Netzwerk für Gesundheit, Klinikum Neukölln, Klinik für Thoraxchirurgie, Berlin, Germany.
⁹ Institute of Virology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
¹⁰ German Centre for Infection Research (DZIF), Germany.
¹¹ These authors contributed equally to this study.
¹² Institute of Physiology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany wolfgang.kuebler@charite.de.
¹³ The Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Toronto, ON, Canada.
¹⁴ Depts. of Surgery and Physiology, University of Toronto, Toronto, ON, Canada.
¹⁵ German Centre for Cardiovascular Research (DZHK), Germany.

PMID: 33154030
PMCID: PMC7651836
DOI: 10.1183/13993003.02384-2020

Plasma mediators in patients with severe COVID-19 cause lung endothelial barrier failure

Laura Michalick et al. Eur Respir J. 2021.

. 2021 Mar 11;57(3):2002384.

doi: 10.1183/13993003.02384-2020. Print 2021 Mar.

Authors

Affiliations

¹ Institute of Physiology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
² Dept of Infectious Diseases and Respiratory Medicine, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
³ Berlin Institute of Health (BIH), Berlin, Germany.
⁴ Max Delbrück Center for Molecular Medicine, Berlin, Germany.
⁵ German Center for Lung Research (DZL), Germany.
⁶ Dept of General, Visceral, Vascular and Thoracic Surgery, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
⁷ Lungenklinik Heckeshorn, HELIOS Klinikum Emil von Behring, Berlin, Germany.
⁸ Vivantes Netzwerk für Gesundheit, Klinikum Neukölln, Klinik für Thoraxchirurgie, Berlin, Germany.
⁹ Institute of Virology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany.
¹⁰ German Centre for Infection Research (DZIF), Germany.
¹¹ These authors contributed equally to this study.
¹² Institute of Physiology, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health (BIH), Berlin, Germany wolfgang.kuebler@charite.de.
¹³ The Keenan Research Centre for Biomedical Science, St. Michael's Hospital, Toronto, ON, Canada.
¹⁴ Depts. of Surgery and Physiology, University of Toronto, Toronto, ON, Canada.
¹⁵ German Centre for Cardiovascular Research (DZHK), Germany.

PMID: 33154030
PMCID: PMC7651836
DOI: 10.1183/13993003.02384-2020

Abstract

The plasma of COVID-19 patients induces pulmonary microvascular barrier failure, which increases with disease severity. Here, a screening platform to test for plasma mediators and the therapeutic potential of barrier stabilising compounds is reported. https://bit.ly/3k4C0tB

PubMed Disclaimer

Conflict of interest statement

Conflict of interest: S. Weidenfeld has nothing to disclose. Conflict of interest: B. Grimmer has nothing to disclose. Conflict of interest: D. Fatykhova has nothing to disclose. Conflict of interest: P.D. Solymosi has nothing to disclose. Conflict of interest: F. Behrens has nothing to disclose. Conflict of interest: M. Dohmen has nothing to disclose. Conflict of interest: M.C. Brack has nothing to disclose. Conflict of interest: S. Schulz has nothing to disclose. Conflict of interest: E. Thomasch has nothing to disclose. Conflict of interest: S. Simmons has nothing to disclose. Conflict of interest: H. Müller-Redetzky has nothing to disclose. Conflict of interest: N. Suttorp has nothing to disclose. Conflict of interest: F. Kurth has nothing to disclose. Conflict of interest: J. Neudecker has nothing to disclose. Conflict of interest: M. Toennies has nothing to disclose. Conflict of interest: T.T. Bauer has nothing to disclose. Conflict of interest: S. Eggeling has nothing to disclose. Conflict of interest: V.M. Corman has nothing to disclose. Conflict of interest: A.C. Hocke has nothing to disclose. Conflict of interest: M. Witzenrath reports grants and personal fees from Actelion, Bayer Health Care, Biotest, Boehringer Ingelheim, Noxxon, Pantherna, Silence Therapeutics and Vaxxilon, grants from Quark Pharma and Takeda Pharma, personal fees from Aptarion, AstraZeneca, Berlin Chemie, Chiesi, GlaxoSmithKline, Novartis, Sinoxa and Teva, outside the submitted work; and has a patent “Means for inhibiting the expression of ANG2 US8829179B2” issued. Conflict of interest: S. Hippenstiel has nothing to disclose. Conflict of interest: W.M. Kuebler reports grants from Ministry of Education and Research (BMBF), German Research Foundation (DFG), German Centre for Cardiovascular Research (DZHK) and Canadian Institutes for Health Resarch (CIHR), during the conduct of the study. Conflict of interest: L. Michalick has nothing to disclose.

Figures

**FIGURE 1**
Plasma of coronavirus disease 2019 (COVID-19) patients induces endothelial barrier failure. a) Barrier integrity of pulmonary microvascular endothelial cell monolayers assessed by electric cell impedance sensing (ECIS) in response to stimulation with 10% (v/v) plasma (at t=0 h) of healthy controls (number of samples/donors: 17/14), and moderate (25/14) or severe (22/16) COVID-19 patients. In cases of multiple samples from the same patient these were obtained at different time-points. Data are normalised to mean endothelial cell (EC) resistance in healthy donors. b) Trans-endothelial electrical resistance (TEER) response of EC monolayers to stimulation with 10% (v/v) plasma (at t=0 h) of healthy controls (7/7), and moderate (4/4), or severe (8/7) COVID-19 patients, with medium control and platelet activating factor as negative and positive controls. c) Representative immunofluorescence images of EC monolayers treated for 6 h with 10% (v/v) plasma of either healthy donors or severe COVID-19 patients stained for (left) VE-cadherin (cyan) or (centre) F-actin (green). Nuclei are shown in blue (DAPI). Gap area per cell (white in F-actin images) was identified by an automated segmentation algorithm (selecting pixels with zero values in both the F-Actin and VE-cadherin channel) using Fiji/MorpholibJ-Plugin, and was quantified for EC monolayers treated with plasma from healthy donors (7/7) or severe COVID-19 patients (7/7) (right). d) Representative western blots and quantification of VE-cadherin and occludin protein levels (normalised to healthy donors) in human lung samples from five donors treated for 24 h with plasma of three healthy donors or five severe COVID-19 patients. *: p≤0.05.

See this image and copyright information in PMC

References

1. Xu Z, Shi L, Wang Y, et al. . Pathological findings of COVID-19 associated with acute respiratory distress syndrome. Lancet Respir Med 2020; 8: 420–422. doi:10.1016/S2213-2600(20)30076-X - DOI - PMC - PubMed
1. Ackermann M, Verleden SE, Kuehnel M, et al. . Pulmonary vascular endothelialitis, thrombosis, and angiogenesis in Covid-19. N Engl J Med 2020; 383: 120–128. doi:10.1056/NEJMoa2015432 - DOI - PMC - PubMed
1. To KF, Lo AWI. Exploring the pathogenesis of severe acute respiratory syndrome (SARS): the tissue distribution of the coronavirus (SARS-CoV) and its putative receptor, angiotensin-converting enzyme 2 (ACE2). J Pathol 2004; 203: 740–743. doi:10.1002/path.1597 - DOI - PMC - PubMed
1. Messner CB, Demichev V, Wendisch D, et al. . Ultra-high-throughput clinical proteomics reveals classifiers of COVID-19 infection. Cell Syst 2020; 11: 11–24.e4. doi:10.1016/j.cels.2020.05.012 - DOI - PMC - PubMed
1. Kuebler WM, Jordt S-E, Liedtke WB. Urgent reconsideration of lung edema as a preventable outcome in COVID-19: inhibition of TRPV4 represents a promising and feasible approach. Am J Physiol Lung Cell Mol Physiol 2020; 318: L1239–L1243. doi:10.1152/ajplung.00161.2020 - DOI - PMC - PubMed

Publication types

Actions

MeSH terms

Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions
Actions

Substances

Actions
Actions
Actions
Actions
Actions

LinkOut - more resources

Full Text Sources
Medical
- MedlinePlus Health Information
Miscellaneous
- NCI CPTAC Assay Portal

Save citation to file

Email citation

Add to Collections

Add to My Bibliography

Your saved search

Create a file for external citation management software

Your RSS Feed

Plasma mediators in patients with severe COVID-19 cause lung endothelial barrier failure

Affiliations

Plasma mediators in patients with severe COVID-19 cause lung endothelial barrier failure

Authors

Affiliations

Abstract

Conflict of interest statement

Figures

References

Publication types

MeSH terms

Substances

LinkOut - more resources

Full Text Sources

Medical

Miscellaneous