Alterations in platelet proteome signature and impaired platelet integrin αIIbβ3 activation in patients with COVID-19

Lucy Goudswaard; Christopher M Williams; Jawad S Khalil; Kate L Burley; Fergus W Hamilton; David T Arnold; Alice Milne; Phil Lewis; Kate J Heesom; Stuart J Mundell; Andrew D Davidson; Alastair W Poole; Ingeborg Hers

doi:10.1101/2022.10.12.511145

Alterations in platelet proteome signature and impaired platelet integrin α_IIbβ₃ activation in patients with COVID-19

Lucy Goudswaard, Christopher M Williams, Jawad S Khalil, Kate L Burley, Fergus W Hamilton, David T Arnold, Alice Milne, Phil Lewis, Kate J Heesom, Stuart J Mundell, Andrew D Davidson, Alastair W Poole, Ingeborg Hers^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

7 Citations (Scopus)

Abstract

Background
Patients with coronavirus disease-19 (COVID-19) are at increased risk of thrombosis, which is associated with altered platelet function and coagulopathy, contributing to excess mortality.

Objectives
We aimed to characterise the mechanism of altered platelet function in COVID-19 patients.
Methods
The platelet proteome, platelet functional responses and platelet-neutrophil aggregates were compared between patients hospitalised with COVID-19 and healthy control subjects using Tandem Mass Tag (TMT) proteomic analysis, Western blotting and flow cytometry.
Results
COVID-19 patients showed a different profile of platelet protein expression (858 altered out of 5773 quantified). Levels of COVID-19 plasma markers were enhanced in COVID-19 platelets. Gene ontology (GO) pathway analysis demonstrated that levels of granule secretory proteins were raised, whereas some platelet activation proteins, such as the thrombopoietin receptor and PKCα, were lowered. Basally, COVID-19 platelets showed enhanced phosphatidylserine (PS) exposure, with unaltered integrin
α_IIbβ₃ activation and P-selectin expression. Agonist-stimulated integrin α_IIbβ₃ activation and PS exposure, but not P-selectin expression, were significantly decreased in COVID-19 patients. COVID-19 patients had high levels of platelet-neutrophil aggregates, even under basal conditions, compared to controls. This interaction was disrupted by blocking P-selectin, demonstrating that platelet P-selectin is critical for the interaction.
Conclusions
Overall, our data suggests the presence of two platelet populations in patients with COVID-19: one with circulating platelets with an altered proteome and reduced functional responses and another with P-selectin expressing neutrophil-associated platelets. Platelet driven thromboinflammation may therefore be one of the key factors enhancing the risk of thrombosis in COVID-19 patients.

Original language	English
Pages (from-to)	1307-1321
Number of pages	15
Journal	Journal of Thrombosis and Haemostasis
Volume	21
Issue number	5
Early online date	27 Jan 2023
DOIs	https://doi.org/10.1101/2022.10.12.511145 https://doi.org/10.1016/j.jtha.2023.01.018
Publication status	Published - 1 May 2023

Bibliographical note

Funding Information:
This work was supported by the University of Bristol Elizabeth Blackwell Institute , the University of Bristol Alumni , the Southmead Hospital Charity , the Wellcome Trust (216277/Z/19/Z and 219472/Z/19/Z), and the British Heart Foundation (FS/17/60/33474, RG/15/16/31758, PG/17/62/33190, PG/21/10760, and SP/F/21/150023). L.J.G. is supported by the BHF Accelerator Award AA/18/1/34219. We thank Borko Amulic and Christopher Rice for their helpful discussions over the observed interactions between platelets and neutrophils. This research was funded in whole, or in part, by the Wellcome Trust. For the purpose of Open Access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission.

Funding Information:
Funding information This work was supported by the University of Bristol Elizabeth Blackwell Institute, the University of Bristol Alumni, the Southmead Hospital Charity , the Wellcome Trust (216277/Z/19/Z and 219472/Z/19/Z), and the British Heart Foundation (FS/17/60/33474, RG/15/16/31758, PG/17/62/33190, PG/21/10760, and SP/F/21/150023). L.J.G. is supported by the BHF Accelerator Award AA/18/1/34219.

Publisher Copyright:
© 2023 The Author(s)

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Academic Respiratory Unit

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Goudswaard, L., Williams, C. M., Khalil, J. S., Burley, K. L., Hamilton, F. W., Arnold, D. T., Milne, A., Lewis, P., Heesom, K. J., Mundell, S. J., Davidson, A. D., Poole, A. W., & Hers, I. (2023). Alterations in platelet proteome signature and impaired platelet integrin α_IIbβ₃ activation in patients with COVID-19. Journal of Thrombosis and Haemostasis, 21(5), 1307-1321. https://doi.org/10.1101/2022.10.12.511145, https://doi.org/10.1016/j.jtha.2023.01.018

@article{7f5f32e2b0b44941906b16184b5e7ece,

title = "Alterations in platelet proteome signature and impaired platelet integrin αIIbβ3 activation in patients with COVID-19",

abstract = "BackgroundPatients with coronavirus disease-19 (COVID-19) are at increased risk of thrombosis, which is associated with altered platelet function and coagulopathy, contributing to excess mortality.ObjectivesWe aimed to characterise the mechanism of altered platelet function in COVID-19 patients.MethodsThe platelet proteome, platelet functional responses and platelet-neutrophil aggregates were compared between patients hospitalised with COVID-19 and healthy control subjects using Tandem Mass Tag (TMT) proteomic analysis, Western blotting and flow cytometry.ResultsCOVID-19 patients showed a different profile of platelet protein expression (858 altered out of 5773 quantified). Levels of COVID-19 plasma markers were enhanced in COVID-19 platelets. Gene ontology (GO) pathway analysis demonstrated that levels of granule secretory proteins were raised, whereas some platelet activation proteins, such as the thrombopoietin receptor and PKCα, were lowered. Basally, COVID-19 platelets showed enhanced phosphatidylserine (PS) exposure, with unaltered integrinαIIbβ3 activation and P-selectin expression. Agonist-stimulated integrin αIIbβ3 activation and PS exposure, but not P-selectin expression, were significantly decreased in COVID-19 patients. COVID-19 patients had high levels of platelet-neutrophil aggregates, even under basal conditions, compared to controls. This interaction was disrupted by blocking P-selectin, demonstrating that platelet P-selectin is critical for the interaction.ConclusionsOverall, our data suggests the presence of two platelet populations in patients with COVID-19: one with circulating platelets with an altered proteome and reduced functional responses and another with P-selectin expressing neutrophil-associated platelets. Platelet driven thromboinflammation may therefore be one of the key factors enhancing the risk of thrombosis in COVID-19 patients.",

author = "Lucy Goudswaard and Williams, {Christopher M} and Khalil, {Jawad S} and Burley, {Kate L} and Hamilton, {Fergus W} and Arnold, {David T} and Alice Milne and Phil Lewis and Heesom, {Kate J} and Mundell, {Stuart J} and Davidson, {Andrew D} and Poole, {Alastair W} and Ingeborg Hers",

note = "Funding Information: This work was supported by the University of Bristol Elizabeth Blackwell Institute , the University of Bristol Alumni , the Southmead Hospital Charity , the Wellcome Trust (216277/Z/19/Z and 219472/Z/19/Z), and the British Heart Foundation (FS/17/60/33474, RG/15/16/31758, PG/17/62/33190, PG/21/10760, and SP/F/21/150023). L.J.G. is supported by the BHF Accelerator Award AA/18/1/34219. We thank Borko Amulic and Christopher Rice for their helpful discussions over the observed interactions between platelets and neutrophils. This research was funded in whole, or in part, by the Wellcome Trust. For the purpose of Open Access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. Funding Information: Funding information This work was supported by the University of Bristol Elizabeth Blackwell Institute, the University of Bristol Alumni, the Southmead Hospital Charity , the Wellcome Trust (216277/Z/19/Z and 219472/Z/19/Z), and the British Heart Foundation (FS/17/60/33474, RG/15/16/31758, PG/17/62/33190, PG/21/10760, and SP/F/21/150023). L.J.G. is supported by the BHF Accelerator Award AA/18/1/34219. Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = may,

day = "1",

doi = "10.1101/2022.10.12.511145",

language = "English",

volume = "21",

pages = "1307--1321",

journal = "Journal of Thrombosis and Haemostasis",

issn = "1538-7933",

publisher = "Wiley-Blackwell",

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}

Goudswaard, L , Williams, CM, Khalil, JS, Burley, KL , Hamilton, FW , Arnold, DT, Milne, A, Lewis, P , Heesom, KJ , Mundell, SJ , Davidson, AD , Poole, AW & Hers, I 2023, 'Alterations in platelet proteome signature and impaired platelet integrin α_IIbβ₃ activation in patients with COVID-19', Journal of Thrombosis and Haemostasis, vol. 21, no. 5, pp. 1307-1321. https://doi.org/10.1101/2022.10.12.511145, https://doi.org/10.1016/j.jtha.2023.01.018

TY - JOUR

T1 - Alterations in platelet proteome signature and impaired platelet integrin αIIbβ3 activation in patients with COVID-19

AU - Goudswaard, Lucy

AU - Williams, Christopher M

AU - Khalil, Jawad S

AU - Burley, Kate L

AU - Hamilton, Fergus W

AU - Arnold, David T

AU - Milne, Alice

AU - Lewis, Phil

AU - Heesom, Kate J

AU - Mundell, Stuart J

AU - Davidson, Andrew D

AU - Poole, Alastair W

AU - Hers, Ingeborg

N1 - Funding Information: This work was supported by the University of Bristol Elizabeth Blackwell Institute , the University of Bristol Alumni , the Southmead Hospital Charity , the Wellcome Trust (216277/Z/19/Z and 219472/Z/19/Z), and the British Heart Foundation (FS/17/60/33474, RG/15/16/31758, PG/17/62/33190, PG/21/10760, and SP/F/21/150023). L.J.G. is supported by the BHF Accelerator Award AA/18/1/34219. We thank Borko Amulic and Christopher Rice for their helpful discussions over the observed interactions between platelets and neutrophils. This research was funded in whole, or in part, by the Wellcome Trust. For the purpose of Open Access, the author has applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. Funding Information: Funding information This work was supported by the University of Bristol Elizabeth Blackwell Institute, the University of Bristol Alumni, the Southmead Hospital Charity , the Wellcome Trust (216277/Z/19/Z and 219472/Z/19/Z), and the British Heart Foundation (FS/17/60/33474, RG/15/16/31758, PG/17/62/33190, PG/21/10760, and SP/F/21/150023). L.J.G. is supported by the BHF Accelerator Award AA/18/1/34219. Publisher Copyright: © 2023 The Author(s)

PY - 2023/5/1

Y1 - 2023/5/1

N2 - BackgroundPatients with coronavirus disease-19 (COVID-19) are at increased risk of thrombosis, which is associated with altered platelet function and coagulopathy, contributing to excess mortality.ObjectivesWe aimed to characterise the mechanism of altered platelet function in COVID-19 patients.MethodsThe platelet proteome, platelet functional responses and platelet-neutrophil aggregates were compared between patients hospitalised with COVID-19 and healthy control subjects using Tandem Mass Tag (TMT) proteomic analysis, Western blotting and flow cytometry.ResultsCOVID-19 patients showed a different profile of platelet protein expression (858 altered out of 5773 quantified). Levels of COVID-19 plasma markers were enhanced in COVID-19 platelets. Gene ontology (GO) pathway analysis demonstrated that levels of granule secretory proteins were raised, whereas some platelet activation proteins, such as the thrombopoietin receptor and PKCα, were lowered. Basally, COVID-19 platelets showed enhanced phosphatidylserine (PS) exposure, with unaltered integrinαIIbβ3 activation and P-selectin expression. Agonist-stimulated integrin αIIbβ3 activation and PS exposure, but not P-selectin expression, were significantly decreased in COVID-19 patients. COVID-19 patients had high levels of platelet-neutrophil aggregates, even under basal conditions, compared to controls. This interaction was disrupted by blocking P-selectin, demonstrating that platelet P-selectin is critical for the interaction.ConclusionsOverall, our data suggests the presence of two platelet populations in patients with COVID-19: one with circulating platelets with an altered proteome and reduced functional responses and another with P-selectin expressing neutrophil-associated platelets. Platelet driven thromboinflammation may therefore be one of the key factors enhancing the risk of thrombosis in COVID-19 patients.

AB - BackgroundPatients with coronavirus disease-19 (COVID-19) are at increased risk of thrombosis, which is associated with altered platelet function and coagulopathy, contributing to excess mortality.ObjectivesWe aimed to characterise the mechanism of altered platelet function in COVID-19 patients.MethodsThe platelet proteome, platelet functional responses and platelet-neutrophil aggregates were compared between patients hospitalised with COVID-19 and healthy control subjects using Tandem Mass Tag (TMT) proteomic analysis, Western blotting and flow cytometry.ResultsCOVID-19 patients showed a different profile of platelet protein expression (858 altered out of 5773 quantified). Levels of COVID-19 plasma markers were enhanced in COVID-19 platelets. Gene ontology (GO) pathway analysis demonstrated that levels of granule secretory proteins were raised, whereas some platelet activation proteins, such as the thrombopoietin receptor and PKCα, were lowered. Basally, COVID-19 platelets showed enhanced phosphatidylserine (PS) exposure, with unaltered integrinαIIbβ3 activation and P-selectin expression. Agonist-stimulated integrin αIIbβ3 activation and PS exposure, but not P-selectin expression, were significantly decreased in COVID-19 patients. COVID-19 patients had high levels of platelet-neutrophil aggregates, even under basal conditions, compared to controls. This interaction was disrupted by blocking P-selectin, demonstrating that platelet P-selectin is critical for the interaction.ConclusionsOverall, our data suggests the presence of two platelet populations in patients with COVID-19: one with circulating platelets with an altered proteome and reduced functional responses and another with P-selectin expressing neutrophil-associated platelets. Platelet driven thromboinflammation may therefore be one of the key factors enhancing the risk of thrombosis in COVID-19 patients.

U2 - 10.1101/2022.10.12.511145

DO - 10.1101/2022.10.12.511145

M3 - Article (Academic Journal)

C2 - 36716966

SN - 1538-7933

VL - 21

SP - 1307

EP - 1321

JO - Journal of Thrombosis and Haemostasis

JF - Journal of Thrombosis and Haemostasis

IS - 5

ER -