S100A8/A9 drives the formation of procoagulant platelets through GPIbα

Martina Colicchia; Waltraud C. Schrottmaier; Gina Perrella; Jasmeet S. Reyat; Jenefa Begum; Alexandre Slater; Joshua Price; Joanne C. Clark; Zhaogong Zhi; Megan J. Simpson; Joshua H. Bourne; Natalie S. Poulter; Abdullah O. Khan; Phillip L.R. Nicolson; Matthew Pugh; Paul Harrison; Asif J. Iqbal; George E. Rainger; Steve P. Watson; Mark R. Thomas; Nicola J. Mutch; Alice Assinger; Julie Rayes

doi:10.1182/blood.2021014966

S100A8/A9 drives the formation of procoagulant platelets through GPIbα

Martina Colicchia, Waltraud C. Schrottmaier, Gina Perrella, Jasmeet S. Reyat, Jenefa Begum, Alexandre Slater, Joshua Price, Joanne C. Clark, Zhaogong Zhi, Megan J. Simpson, Joshua H. Bourne, Natalie S. Poulter, Abdullah O. Khan, Phillip L.R. Nicolson, Matthew Pugh, Paul Harrison, Asif J. Iqbal, George E. Rainger, Steve P. Watson, Mark R. ThomasNicola J. Mutch, Alice Assinger, Julie Rayes^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

35 Citations (Scopus)

Abstract

S100A8/A9, also known as “calprotectin” or “MRP8/14,” is an alarmin primarily secreted by activated myeloid cells with antimicrobial, proinflammatory, and prothrombotic properties. Increased plasma levels of S100A8/A9 in thrombo-inflammatory diseases are associated with thrombotic complications. We assessed the presence of S100A8/A9 in the plasma and lung autopsies from patients with COVID-19 and investigated the molecular mechanism by which S100A8/A9 affects platelet function and thrombosis. S100A8/A9 plasma levels were increased in patients with COVID-19 and sustained high levels during hospitalization correlated with poor outcomes. Heterodimeric S100A8/A9 was mainly detected in neutrophils and deposited on the vessel wall in COVID-19 lung autopsies. Immobilization of S100A8/A9 with collagen accelerated the formation of a fibrin-rich network after perfusion of recalcified blood at venous shear. In vitro, platelets adhered and partially spread on S100A8/A9, leading to the formation of distinct populations of either P-selectin or phosphatidylserine (PS)-positive platelets. By using washed platelets, soluble S100A8/A9 induced PS exposure but failed to induce platelet aggregation, despite GPIIb/IIIa activation and alpha-granule secretion. We identified GPIbα as the receptor for S100A8/A9 on platelets inducing the formation of procoagulant platelets with a supporting role for CD36. The effect of S100A8/A9 on platelets was abolished by recombinant GPIbα ectodomain, platelets from a patient with Bernard-Soulier syndrome with GPIb-IX-V deficiency, and platelets from mice deficient in the extracellular domain of GPIbα. We identified the S100A8/A9-GPIbα axis as a novel targetable prothrombotic pathway inducing procoagulant platelets and fibrin formation, in particular in diseases associated with high levels of S100A8/A9, such as COVID-19.

Original language	English
Pages (from-to)	2626-2643
Number of pages	18
Journal	Blood
Volume	140
Issue number	24
DOIs	https://doi.org/10.1182/blood.2021014966
Publication status	Published - 15 Dec 2022

Bibliographical note

Funding Information:
J.R. holds a British Heart Foundation (BHF) Intermediate Fellowship (FS/IBSRF/20/25039). This research was partially supported by a BHF project grant (PG/21/10737), the BHF Accelerator Award (AA/18/2/34218), UK Spine Knowledge exchange (R78606/CN003), and the Medical Research Council (Grant Number MC_PC_19029) for J.R. M.C. is supported by the Wellcome Trust 4 Year PhD studentship program on Mechanisms of Inflammatory Disease (204951). A.A. is supported by the Austrian Science Fund (P32064 and P34783). G.P. is supported by a Birmingham-Maastricht studentship. S.P.W. holds a BHF Chair (CH03/003). A.O.K. is a Henry Wellcome fellow (218649/Z/19/Z). N.J.M. is supported by University of Aberdeen Development Trust and National Health Service Grampian Endowment funds (COV19-004 and 20/021). A.J.I. holds a Birmingham fellowship.

Acknowledgments:
The authors thank Beata Grygielska for genotyping mice and University of Birmingham Enterprise Ltd or the translational Research Team for their support. J.R. holds a British Heart Foundation (BHF) Intermediate Fellowship (FS/IBSRF/20/25039). This research was partially supported by a BHF project grant (PG/21/10737), the BHF Accelerator Award (AA/18/2/34218), UK Spine Knowledge exchange (R78606/CN003), and the Medical Research Council (Grant Number MC_PC_19029) for J.R. M.C. is supported by the Wellcome Trust 4 Year PhD studentship program on Mechanisms of Inflammatory Disease (204951). A.A. is supported by the Austrian Science Fund (P32064 and P34783). G.P. is supported by a Birmingham-Maastricht studentship. S.P.W. holds a BHF Chair (CH03/003). A.O.K. is a Henry Wellcome fellow (218649/Z/19/Z). N.J.M. is supported by University of Aberdeen Development Trust and National Health Service Grampian Endowment funds (COV19-004 and 20/021). A.J.I. holds a Birmingham fellowship. Contribution: M.C. designed and performed experiments, collected and analyzed data, and wrote the manuscript; W.C.S. G.P. M.J.S. J.P. J.C.C. J.B. J.S.R. Z.Z. and A.S. performed experiments and analyzed data; J.H.B. and A.O.K. analyzed data; M.P. N.S.P. P.L.R.N. P.H. A.J.I. G.E.R. S.P.W. and M.R.T. provided key reagents and contributed to data analysis; N.J.M. and A.A. provided key reagents and supported research analysis; J.R. designed research and experiments, performed experiments, collected and analyzed data, and wrote the manuscript; and all authors read and approved the paper.

Keywords

S100A8/A9
calprotectin
fibrin
procoagulant platelets
GPIbα
Calgranulin A/metabolism
Phosphatidylserines/metabolism
Fibrin/metabolism
Blood Platelets/metabolism
Platelet Aggregation
Animals
COVID-19/metabolism
Mice

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Colicchia, M., Schrottmaier, W. C., Perrella, G., Reyat, J. S., Begum, J., Slater, A., Price, J., Clark, J. C., Zhi, Z., Simpson, M. J., Bourne, J. H., Poulter, N. S., Khan, A. O., Nicolson, P. L. R., Pugh, M., Harrison, P., Iqbal, A. J., Rainger, G. E., Watson, S. P., ... Rayes, J. (2022). S100A8/A9 drives the formation of procoagulant platelets through GPIbα. Blood, 140(24), 2626-2643. https://doi.org/10.1182/blood.2021014966

Colicchia, M, Schrottmaier, WC, Perrella, G, Reyat, JS, Begum, J, Slater, A, Price, J, Clark, JC, Zhi, Z, Simpson, MJ, Bourne, JH, Poulter, NS, Khan, AO, Nicolson, PLR, Pugh, M, Harrison, P, Iqbal, AJ, Rainger, GE, Watson, SP, Thomas, MR, Mutch, NJ, Assinger, A & Rayes, J 2022, 'S100A8/A9 drives the formation of procoagulant platelets through GPIbα', Blood, vol. 140, no. 24, pp. 2626-2643. https://doi.org/10.1182/blood.2021014966

@article{e82dad601fa645e4966e8a134d97e254,

title = "S100A8/A9 drives the formation of procoagulant platelets through GPIbα",

abstract = "S100A8/A9, also known as “calprotectin” or “MRP8/14,” is an alarmin primarily secreted by activated myeloid cells with antimicrobial, proinflammatory, and prothrombotic properties. Increased plasma levels of S100A8/A9 in thrombo-inflammatory diseases are associated with thrombotic complications. We assessed the presence of S100A8/A9 in the plasma and lung autopsies from patients with COVID-19 and investigated the molecular mechanism by which S100A8/A9 affects platelet function and thrombosis. S100A8/A9 plasma levels were increased in patients with COVID-19 and sustained high levels during hospitalization correlated with poor outcomes. Heterodimeric S100A8/A9 was mainly detected in neutrophils and deposited on the vessel wall in COVID-19 lung autopsies. Immobilization of S100A8/A9 with collagen accelerated the formation of a fibrin-rich network after perfusion of recalcified blood at venous shear. In vitro, platelets adhered and partially spread on S100A8/A9, leading to the formation of distinct populations of either P-selectin or phosphatidylserine (PS)-positive platelets. By using washed platelets, soluble S100A8/A9 induced PS exposure but failed to induce platelet aggregation, despite GPIIb/IIIa activation and alpha-granule secretion. We identified GPIbα as the receptor for S100A8/A9 on platelets inducing the formation of procoagulant platelets with a supporting role for CD36. The effect of S100A8/A9 on platelets was abolished by recombinant GPIbα ectodomain, platelets from a patient with Bernard-Soulier syndrome with GPIb-IX-V deficiency, and platelets from mice deficient in the extracellular domain of GPIbα. We identified the S100A8/A9-GPIbα axis as a novel targetable prothrombotic pathway inducing procoagulant platelets and fibrin formation, in particular in diseases associated with high levels of S100A8/A9, such as COVID-19.",

keywords = "S100A8/A9, calprotectin, fibrin, procoagulant platelets, GPIbα, Calgranulin A/metabolism, Phosphatidylserines/metabolism, Fibrin/metabolism, Blood Platelets/metabolism, Platelet Aggregation, Animals, COVID-19/metabolism, Mice",

author = "Martina Colicchia and Schrottmaier, {Waltraud C.} and Gina Perrella and Reyat, {Jasmeet S.} and Jenefa Begum and Alexandre Slater and Joshua Price and Clark, {Joanne C.} and Zhaogong Zhi and Simpson, {Megan J.} and Bourne, {Joshua H.} and Poulter, {Natalie S.} and Khan, {Abdullah O.} and Nicolson, {Phillip L.R.} and Matthew Pugh and Paul Harrison and Iqbal, {Asif J.} and Rainger, {George E.} and Watson, {Steve P.} and Thomas, {Mark R.} and Mutch, {Nicola J.} and Alice Assinger and Julie Rayes",

note = "Funding Information: J.R. holds a British Heart Foundation (BHF) Intermediate Fellowship (FS/IBSRF/20/25039). This research was partially supported by a BHF project grant (PG/21/10737), the BHF Accelerator Award (AA/18/2/34218), UK Spine Knowledge exchange (R78606/CN003), and the Medical Research Council (Grant Number MC_PC_19029) for J.R. M.C. is supported by the Wellcome Trust 4 Year PhD studentship program on Mechanisms of Inflammatory Disease (204951). A.A. is supported by the Austrian Science Fund (P32064 and P34783). G.P. is supported by a Birmingham-Maastricht studentship. S.P.W. holds a BHF Chair (CH03/003). A.O.K. is a Henry Wellcome fellow (218649/Z/19/Z). N.J.M. is supported by University of Aberdeen Development Trust and National Health Service Grampian Endowment funds (COV19-004 and 20/021). A.J.I. holds a Birmingham fellowship. Acknowledgments: The authors thank Beata Grygielska for genotyping mice and University of Birmingham Enterprise Ltd or the translational Research Team for their support. J.R. holds a British Heart Foundation (BHF) Intermediate Fellowship (FS/IBSRF/20/25039). This research was partially supported by a BHF project grant (PG/21/10737), the BHF Accelerator Award (AA/18/2/34218), UK Spine Knowledge exchange (R78606/CN003), and the Medical Research Council (Grant Number MC_PC_19029) for J.R. M.C. is supported by the Wellcome Trust 4 Year PhD studentship program on Mechanisms of Inflammatory Disease (204951). A.A. is supported by the Austrian Science Fund (P32064 and P34783). G.P. is supported by a Birmingham-Maastricht studentship. S.P.W. holds a BHF Chair (CH03/003). A.O.K. is a Henry Wellcome fellow (218649/Z/19/Z). N.J.M. is supported by University of Aberdeen Development Trust and National Health Service Grampian Endowment funds (COV19-004 and 20/021). A.J.I. holds a Birmingham fellowship. Contribution: M.C. designed and performed experiments, collected and analyzed data, and wrote the manuscript; W.C.S. G.P. M.J.S. J.P. J.C.C. J.B. J.S.R. Z.Z. and A.S. performed experiments and analyzed data; J.H.B. and A.O.K. analyzed data; M.P. N.S.P. P.L.R.N. P.H. A.J.I. G.E.R. S.P.W. and M.R.T. provided key reagents and contributed to data analysis; N.J.M. and A.A. provided key reagents and supported research analysis; J.R. designed research and experiments, performed experiments, collected and analyzed data, and wrote the manuscript; and all authors read and approved the paper. ",

year = "2022",

month = dec,

day = "15",

doi = "10.1182/blood.2021014966",

language = "English",

volume = "140",

pages = "2626--2643",

journal = "Blood",

issn = "0006-4971",

publisher = "American Society of Hematology",

number = "24",

}

TY - JOUR

T1 - S100A8/A9 drives the formation of procoagulant platelets through GPIbα

AU - Colicchia, Martina

AU - Schrottmaier, Waltraud C.

AU - Perrella, Gina

AU - Reyat, Jasmeet S.

AU - Begum, Jenefa

AU - Slater, Alexandre

AU - Price, Joshua

AU - Clark, Joanne C.

AU - Zhi, Zhaogong

AU - Simpson, Megan J.

AU - Bourne, Joshua H.

AU - Poulter, Natalie S.

AU - Khan, Abdullah O.

AU - Nicolson, Phillip L.R.

AU - Pugh, Matthew

AU - Harrison, Paul

AU - Iqbal, Asif J.

AU - Rainger, George E.

AU - Watson, Steve P.

AU - Thomas, Mark R.

AU - Mutch, Nicola J.

AU - Assinger, Alice

AU - Rayes, Julie

N1 - Funding Information: J.R. holds a British Heart Foundation (BHF) Intermediate Fellowship (FS/IBSRF/20/25039). This research was partially supported by a BHF project grant (PG/21/10737), the BHF Accelerator Award (AA/18/2/34218), UK Spine Knowledge exchange (R78606/CN003), and the Medical Research Council (Grant Number MC_PC_19029) for J.R. M.C. is supported by the Wellcome Trust 4 Year PhD studentship program on Mechanisms of Inflammatory Disease (204951). A.A. is supported by the Austrian Science Fund (P32064 and P34783). G.P. is supported by a Birmingham-Maastricht studentship. S.P.W. holds a BHF Chair (CH03/003). A.O.K. is a Henry Wellcome fellow (218649/Z/19/Z). N.J.M. is supported by University of Aberdeen Development Trust and National Health Service Grampian Endowment funds (COV19-004 and 20/021). A.J.I. holds a Birmingham fellowship. Acknowledgments: The authors thank Beata Grygielska for genotyping mice and University of Birmingham Enterprise Ltd or the translational Research Team for their support. J.R. holds a British Heart Foundation (BHF) Intermediate Fellowship (FS/IBSRF/20/25039). This research was partially supported by a BHF project grant (PG/21/10737), the BHF Accelerator Award (AA/18/2/34218), UK Spine Knowledge exchange (R78606/CN003), and the Medical Research Council (Grant Number MC_PC_19029) for J.R. M.C. is supported by the Wellcome Trust 4 Year PhD studentship program on Mechanisms of Inflammatory Disease (204951). A.A. is supported by the Austrian Science Fund (P32064 and P34783). G.P. is supported by a Birmingham-Maastricht studentship. S.P.W. holds a BHF Chair (CH03/003). A.O.K. is a Henry Wellcome fellow (218649/Z/19/Z). N.J.M. is supported by University of Aberdeen Development Trust and National Health Service Grampian Endowment funds (COV19-004 and 20/021). A.J.I. holds a Birmingham fellowship. Contribution: M.C. designed and performed experiments, collected and analyzed data, and wrote the manuscript; W.C.S. G.P. M.J.S. J.P. J.C.C. J.B. J.S.R. Z.Z. and A.S. performed experiments and analyzed data; J.H.B. and A.O.K. analyzed data; M.P. N.S.P. P.L.R.N. P.H. A.J.I. G.E.R. S.P.W. and M.R.T. provided key reagents and contributed to data analysis; N.J.M. and A.A. provided key reagents and supported research analysis; J.R. designed research and experiments, performed experiments, collected and analyzed data, and wrote the manuscript; and all authors read and approved the paper.

PY - 2022/12/15

Y1 - 2022/12/15

N2 - S100A8/A9, also known as “calprotectin” or “MRP8/14,” is an alarmin primarily secreted by activated myeloid cells with antimicrobial, proinflammatory, and prothrombotic properties. Increased plasma levels of S100A8/A9 in thrombo-inflammatory diseases are associated with thrombotic complications. We assessed the presence of S100A8/A9 in the plasma and lung autopsies from patients with COVID-19 and investigated the molecular mechanism by which S100A8/A9 affects platelet function and thrombosis. S100A8/A9 plasma levels were increased in patients with COVID-19 and sustained high levels during hospitalization correlated with poor outcomes. Heterodimeric S100A8/A9 was mainly detected in neutrophils and deposited on the vessel wall in COVID-19 lung autopsies. Immobilization of S100A8/A9 with collagen accelerated the formation of a fibrin-rich network after perfusion of recalcified blood at venous shear. In vitro, platelets adhered and partially spread on S100A8/A9, leading to the formation of distinct populations of either P-selectin or phosphatidylserine (PS)-positive platelets. By using washed platelets, soluble S100A8/A9 induced PS exposure but failed to induce platelet aggregation, despite GPIIb/IIIa activation and alpha-granule secretion. We identified GPIbα as the receptor for S100A8/A9 on platelets inducing the formation of procoagulant platelets with a supporting role for CD36. The effect of S100A8/A9 on platelets was abolished by recombinant GPIbα ectodomain, platelets from a patient with Bernard-Soulier syndrome with GPIb-IX-V deficiency, and platelets from mice deficient in the extracellular domain of GPIbα. We identified the S100A8/A9-GPIbα axis as a novel targetable prothrombotic pathway inducing procoagulant platelets and fibrin formation, in particular in diseases associated with high levels of S100A8/A9, such as COVID-19.

AB - S100A8/A9, also known as “calprotectin” or “MRP8/14,” is an alarmin primarily secreted by activated myeloid cells with antimicrobial, proinflammatory, and prothrombotic properties. Increased plasma levels of S100A8/A9 in thrombo-inflammatory diseases are associated with thrombotic complications. We assessed the presence of S100A8/A9 in the plasma and lung autopsies from patients with COVID-19 and investigated the molecular mechanism by which S100A8/A9 affects platelet function and thrombosis. S100A8/A9 plasma levels were increased in patients with COVID-19 and sustained high levels during hospitalization correlated with poor outcomes. Heterodimeric S100A8/A9 was mainly detected in neutrophils and deposited on the vessel wall in COVID-19 lung autopsies. Immobilization of S100A8/A9 with collagen accelerated the formation of a fibrin-rich network after perfusion of recalcified blood at venous shear. In vitro, platelets adhered and partially spread on S100A8/A9, leading to the formation of distinct populations of either P-selectin or phosphatidylserine (PS)-positive platelets. By using washed platelets, soluble S100A8/A9 induced PS exposure but failed to induce platelet aggregation, despite GPIIb/IIIa activation and alpha-granule secretion. We identified GPIbα as the receptor for S100A8/A9 on platelets inducing the formation of procoagulant platelets with a supporting role for CD36. The effect of S100A8/A9 on platelets was abolished by recombinant GPIbα ectodomain, platelets from a patient with Bernard-Soulier syndrome with GPIb-IX-V deficiency, and platelets from mice deficient in the extracellular domain of GPIbα. We identified the S100A8/A9-GPIbα axis as a novel targetable prothrombotic pathway inducing procoagulant platelets and fibrin formation, in particular in diseases associated with high levels of S100A8/A9, such as COVID-19.

KW - S100A8/A9

KW - calprotectin

KW - fibrin

KW - procoagulant platelets

KW - GPIbα

KW - Calgranulin A/metabolism

KW - Phosphatidylserines/metabolism

KW - Fibrin/metabolism

KW - Blood Platelets/metabolism

KW - Platelet Aggregation

KW - Animals

KW - COVID-19/metabolism

KW - Mice

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U2 - 10.1182/blood.2021014966

DO - 10.1182/blood.2021014966

M3 - Article

C2 - 36026606

AN - SCOPUS:85141298221

SN - 0006-4971

VL - 140

SP - 2626

EP - 2643

JO - Blood

JF - Blood

IS - 24

ER -

S100A8/A9 drives the formation of procoagulant platelets through GPIbα

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Keywords

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