Abstract
Background: Alterations in platelet function have been implicated in the pathophysiology of COVID-19 since the beginning of the pandemic. While early reports linked hyperactivated platelets to thromboembolic events in COVID-19, subsequent investigations demonstrated hyporeactive platelets with a procoagulant phenotype. Mitochondria are important for energy metabolism and the function of platelets. Objectives: Here, we sought to map the energy metabolism of platelets in a cohort of noncritically ill COVID-19 patients and assess platelet mitochondrial function, activation status, and responsiveness to external stimuli. Methods: We enrolled hospitalized COVID-19 patients and controls between October 2020 and December 2021. Platelets function and metabolism was analyzed by flow cytometry, metabolomics, glucose fluxomics, electron and fluorescence microscopy and western blot. Results: Platelets from COVID-19 patients showed increased phosphatidylserine externalization indicating a procoagulant phenotype and hyporeactivity to ex vivo stimuli, associated with profound mitochondrial dysfunction characterized by mitochondrial depolarization, lower mitochondrial DNA–encoded transcript levels, an altered mitochondrial morphology consistent with increased mitochondrial fission, and increased pyruvate/lactate ratios in platelet supernatants. Metabolic profiling by untargeted metabolomics revealed NADH, NAD+, and ATP among the top decreased metabolites in patients’ platelets, suggestive of energy metabolism failure. Consistently, platelet fluxomics analyses showed a strongly reduced utilization of 13C-glucose in all major energy pathways together with a rerouting of glucose to de novo generation of purine metabolites. Patients’ platelets further showed evidence of oxidative stress, together with increased glutathione oxidation and synthesis. Addition of plasma from COVID-19 patients to normal platelets partially reproduced the phenotype of patients’ platelets and disclosed a temporal relationship between mitochondrial decay and (subsequent) phosphatidylserine exposure and hyporeactivity. Conclusion: These data link energy metabolism failure in platelets from COVID-19 patients with a prothrombotic platelet phenotype with features matching cell death.
Original language | English |
---|---|
Article number | 102213 |
Journal | Research and practice in thrombosis and haemostasis |
Volume | 7 |
Issue number | 7 |
DOIs | |
Publication status | Published - Oct 2023 |
Keywords
- COVID-19
- blood platelets
- cell death
- energy metabolism
- mitochondria
- oxidative stress
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In: Research and practice in thrombosis and haemostasis, Vol. 7, No. 7, 102213, 10.2023.
Research output: Contribution to journal › Article › Academic › peer-review
TY - JOUR
T1 - Platelets of COVID-19 patients display mitochondrial dysfunction, oxidative stress, and energy metabolism failure compatible with cell death
AU - Amsterdam University Medical Center COVID-19 Biobank Study Group
AU - Léopold, Valentine
AU - Chouchane, Osoul
AU - Butler, Joe M.
AU - Schuurman, Alex R.
AU - Michels, Erik H.A.
AU - de Brabander, Justin
AU - Schomakers, Bauke V.
AU - van Weeghel, Michel
AU - Picavet-Havik, Daisy I.
AU - Grootemaat, Anita E.
AU - Douma, Renée A.
AU - Reijnders, Tom
AU - Klarenbeek, Augustijn M.
AU - Appelman, Brent
AU - Agtmael, Michiel van
AU - Algera, Anne Geke
AU - van Baarle, Floor
AU - Beudel, Martijn
AU - Bogaard, Harm Jan
AU - Bomers, Marije
AU - Bonta, Peter
AU - Bos, Lieuwe
AU - Botta, Michela
AU - de Bree, Godelieve
AU - de Bruin, Sanne
AU - Bugiani, Marianna
AU - Bulle, Esther
AU - Buis, David T.P.
AU - Clohert, Alex
AU - Dijkstra, Mirjam
AU - Dongelmans, Dave A.
AU - Dujardin, Romein W.G.
AU - Elbers, Paul
AU - Fleuren, Lucas
AU - Geerlings, Suzanne
AU - Geijtenbeek, Theo
AU - Girbes, Armand
AU - Goorhuis, Bram
AU - Grobusch, Martin P.
AU - Hemke, Robert
AU - Hermans, Sabine M.
AU - Lim, Endry H.T.
AU - Nossent, Esther J.
AU - Peters, Edgar
AU - Sigaloff, Kim
AU - Teunissen, Charlotte
AU - Thoral, Patrick
AU - Tuinman, Pieter R.
AU - de Vries, Heder
AU - Houtkooper, Riekelt H.
AU - Amsterdam UMC Covid-19 Biobank
AU - Harris, Vanessa
AU - Hollmann, Markus
AU - Horn, Janneke
AU - Hovius, Joppe W.
AU - de Jong, Menno D.
AU - Paulus, Frederique
AU - van der Poll, Tom
AU - Raasveld, Jorinde
AU - de Rotte, Maurits C.F.J.
AU - Schinkel, Michiel
AU - Schultz, Marcus J.
AU - Stijnis, Cornelis S.
AU - van der Valk, Marc
AU - Veelo, Denise
AU - Zwinderman, A. H.
AU - Brouwer, Matthijs C.
AU - Vlaar, Alexander P.J.
AU - van de Beek, Diederik
AU - van der Wel, Nicole N.
AU - den Dunnen, Jeroen
AU - van't Veer, Cornelis
N1 - Funding Information: V.L. was funded by Fondation Bettencourt-Schueller and Institut Servier , O.C. by Landsteiner Foundation ( LSBR # 1901), J.M.B by European Union’s Horizon 2020 program (grant agreement 847422 – ImmunoSep), E.H.A.M. and J.d.B. by European Union’s Horizon 2020 program (grant agreement 847786 – FAIR). Funding Information: Mark Dings for providing reagents, Danielle Kruijswijk and Fayola de Lange for their technical assistance and laboratory guidance. Amsterdam UMC COVID-19 biobank: Michiel van Agtmael2, Anne Geke Algera1, Brent Appelman2, Floor van Baarle1, Martijn Beudel4, Harm Jan Bogaard5, Marije Bomers2, Peter Bonta5, Lieuwe Bos1, Michela Botta1, Justin de Brabander2, Godelieve de Bree2, Sanne de Bruin1, Marianna Bugiani5, Esther Bulle1, David T.P. Buis1, Osoul Chouchane2, Alex Clohert3, Mirjam Dijkstra12, Dave A. Dongelmans1, Romein W.G. Dujardin1, Paul Elbers1, Lucas Fleuren1, Suzanne Geerlings2, Theo Geijtenbeek3, Armand Girbes1, Bram Goorhuis2, Martin P. Grobusch2, Laura Hagens1, Jorg Hamann7, Vanessa Harris2, Robert Hemke8, Sabine M. Hermans2, Leo Heunks1, Markus Hollmann6, Janneke Horn1, Joppe W. Hovius2, Menno D. de Jong9, Rutger Koning4, Endry H.T. Lim1, Niels van Mourik1, Jeaninne Nellen2, Esther J. Nossent5, Sabine Olie4, Frederique Paulus1, Edgar Peters2, Dan A.I. Pina-Fuentes4, Tom van der Poll2, Bennedikt Preckel6, Jorinde Raasveld1, Tom Reijnders2, Maurits C.F.J. de Rotte12, Michiel Schinkel2, Marcus J. Schultz1, Femke A.P. Schrauwen12, Alex Schuurman2, Jaap Schuurmans1, Kim Sigaloff1, Marleen A. Slim1,2, Patrick Smeele5, Marry Smit1, Cornelis S. Stijnis2, Willemke Stilma1, Charlotte Teunissen11, Patrick Thoral1, Anissa M. Tsonas1, Pieter R. Tuinman1, Marc van der Valk2, Denise Veelo6, Carolien Volleman1, Heder de Vries1, Lonneke A. Vught1,2, Michèle van Vugt2, Dorien Wouters12, A. H. Zwinderman13, Matthijs C. Brouwer4, W. Joost Wiersinga2, Alexander P.J. Vlaar1, Diederik van de Beek (d.vandebeek@amsterdamumc.nl)4. 1Department of Intensive Care, Amsterdam UMC, Amsterdam, The Netherlands; 2Department of Infectious Diseases, Amsterdam UMC, Amsterdam, The Netherlands; 3Experimental Immunology, Amsterdam UMC, Amsterdam, The Netherlands; 4Department of Neurology, Amsterdam UMC, Amsterdam Neuroscience, Amsterdam, The Netherlands; 5Department of Pulmonology, Amsterdam UMC, Amsterdam, The Netherlands; 6Department of Anesthesiology, Amsterdam UMC, Amsterdam, The Netherlands; 7Amsterdam UMC Biobank Core Facility, Amsterdam UMC, Amsterdam, The Netherlands; 8Department of Radiology, Amsterdam UMC, Amsterdam, The Netherlands; 9Department of Medical Microbiology, Amsterdam UMC, Amsterdam, The Netherlands; 10Department of Internal Medicine, Amsterdam UMC, Amsterdam, The Netherlands; 11Neurochemical Laboratory, Amsterdam UMC, Amsterdam, The Netherlands; 12Department of Clinical Chemistry, Amsterdam UMC, Amsterdam, The Netherlands; 13Department of Clinical Epidemiology, Biostatistics and Bioinformatics, Amsterdam UMC, Amsterdam, The Netherlands. V.L. was funded by Fondation Bettencourt-Schueller and Institut Servier, O.C. by Landsteiner Foundation (LSBR # 1901), J.M.B by European Union's Horizon 2020 program (grant agreement 847422 – ImmunoSep), E.H.A.M. and J.d.B. by European Union's Horizon 2020 program (grant agreement 847786 – FAIR). The ELDER-BIOME study was approved by the AMC Ethics Committee (protocol number NL57847.018.16); the Amsterdam UMC COVID-19 biobank study was approved by the Biobank Ethics Committees of both Amsterdam UMC hospitals (AMC and Vrije Universiteit Medical Center; reference number 2020_065). Written informed consent was obtained from all patients and controls. V.L.: conceptualization, investigation, methodology, visualization, writing original draft, review, and editing. O.C.: investigation. J.B. A.R.S. T.D.Y.R.: data curation, software. B.V.S and M.vW.: conceptualization, investigation, data curation, software. D.P. A.E.G. A.M.K.: investigation, resources. E.M. J.dB. R.D. B.A. W.J.W. and Amsterdam Biobank: resources, project administration. Nv.W. J.dD. R.H.: conceptualization, writing review, and editing. Cv.V.: conceptualization, methodology, writing review, and editing, supervision. T.vdP.: funding acquisition, conceptualization, supervision, writing review, and editing. There are no competing interests to disclose. Publisher Copyright: © 2023 The Authors
PY - 2023/10
Y1 - 2023/10
N2 - Background: Alterations in platelet function have been implicated in the pathophysiology of COVID-19 since the beginning of the pandemic. While early reports linked hyperactivated platelets to thromboembolic events in COVID-19, subsequent investigations demonstrated hyporeactive platelets with a procoagulant phenotype. Mitochondria are important for energy metabolism and the function of platelets. Objectives: Here, we sought to map the energy metabolism of platelets in a cohort of noncritically ill COVID-19 patients and assess platelet mitochondrial function, activation status, and responsiveness to external stimuli. Methods: We enrolled hospitalized COVID-19 patients and controls between October 2020 and December 2021. Platelets function and metabolism was analyzed by flow cytometry, metabolomics, glucose fluxomics, electron and fluorescence microscopy and western blot. Results: Platelets from COVID-19 patients showed increased phosphatidylserine externalization indicating a procoagulant phenotype and hyporeactivity to ex vivo stimuli, associated with profound mitochondrial dysfunction characterized by mitochondrial depolarization, lower mitochondrial DNA–encoded transcript levels, an altered mitochondrial morphology consistent with increased mitochondrial fission, and increased pyruvate/lactate ratios in platelet supernatants. Metabolic profiling by untargeted metabolomics revealed NADH, NAD+, and ATP among the top decreased metabolites in patients’ platelets, suggestive of energy metabolism failure. Consistently, platelet fluxomics analyses showed a strongly reduced utilization of 13C-glucose in all major energy pathways together with a rerouting of glucose to de novo generation of purine metabolites. Patients’ platelets further showed evidence of oxidative stress, together with increased glutathione oxidation and synthesis. Addition of plasma from COVID-19 patients to normal platelets partially reproduced the phenotype of patients’ platelets and disclosed a temporal relationship between mitochondrial decay and (subsequent) phosphatidylserine exposure and hyporeactivity. Conclusion: These data link energy metabolism failure in platelets from COVID-19 patients with a prothrombotic platelet phenotype with features matching cell death.
AB - Background: Alterations in platelet function have been implicated in the pathophysiology of COVID-19 since the beginning of the pandemic. While early reports linked hyperactivated platelets to thromboembolic events in COVID-19, subsequent investigations demonstrated hyporeactive platelets with a procoagulant phenotype. Mitochondria are important for energy metabolism and the function of platelets. Objectives: Here, we sought to map the energy metabolism of platelets in a cohort of noncritically ill COVID-19 patients and assess platelet mitochondrial function, activation status, and responsiveness to external stimuli. Methods: We enrolled hospitalized COVID-19 patients and controls between October 2020 and December 2021. Platelets function and metabolism was analyzed by flow cytometry, metabolomics, glucose fluxomics, electron and fluorescence microscopy and western blot. Results: Platelets from COVID-19 patients showed increased phosphatidylserine externalization indicating a procoagulant phenotype and hyporeactivity to ex vivo stimuli, associated with profound mitochondrial dysfunction characterized by mitochondrial depolarization, lower mitochondrial DNA–encoded transcript levels, an altered mitochondrial morphology consistent with increased mitochondrial fission, and increased pyruvate/lactate ratios in platelet supernatants. Metabolic profiling by untargeted metabolomics revealed NADH, NAD+, and ATP among the top decreased metabolites in patients’ platelets, suggestive of energy metabolism failure. Consistently, platelet fluxomics analyses showed a strongly reduced utilization of 13C-glucose in all major energy pathways together with a rerouting of glucose to de novo generation of purine metabolites. Patients’ platelets further showed evidence of oxidative stress, together with increased glutathione oxidation and synthesis. Addition of plasma from COVID-19 patients to normal platelets partially reproduced the phenotype of patients’ platelets and disclosed a temporal relationship between mitochondrial decay and (subsequent) phosphatidylserine exposure and hyporeactivity. Conclusion: These data link energy metabolism failure in platelets from COVID-19 patients with a prothrombotic platelet phenotype with features matching cell death.
KW - COVID-19
KW - blood platelets
KW - cell death
KW - energy metabolism
KW - mitochondria
KW - oxidative stress
UR - http://www.scopus.com/inward/record.url?scp=85178064281&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.rpth.2023.102213
DO - https://doi.org/10.1016/j.rpth.2023.102213
M3 - Article
C2 - 38077825
SN - 2475-0379
VL - 7
JO - Research and practice in thrombosis and haemostasis
JF - Research and practice in thrombosis and haemostasis
IS - 7
M1 - 102213
ER -