Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability

Joanne L. Mitchell; Gemma Little; Alexander P. Bye; Renato S. Gaspar; Amanda J. Unsworth; Neline Kriek; Tanya Sage; Alexander Stainer; Ibidayo Sangowawa; Gael B. Morrow; Ricardo N. Bastos; Susan Shapiro; Michael J.R. Desborough; Nicola Curry; Jonathan M. Gibbins; Claire S. Whyte; Nicola J. Mutch; Christopher I. Jones

doi:10.1016/j.rpth.2023.100200

Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability

Joanne L. Mitchell^*, Gemma Little, Alexander P. Bye, Renato S. Gaspar, Amanda J. Unsworth, Neline Kriek, Tanya Sage, Alexander Stainer, Ibidayo Sangowawa, Gael B. Morrow, Ricardo N. Bastos, Susan Shapiro, Michael J.R. Desborough, Nicola Curry, Jonathan M. Gibbins, Claire S. Whyte, Nicola J. Mutch, Christopher I. Jones

^*Corresponding author for this work

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

3 Citations (Scopus)

Abstract

Background: Factor XIII (FXIII) is an important proenzyme in the hemostatic system. The plasma-derived enzyme activated FXIII cross-links fibrin fibers within thrombi to increase their mechanical strength and cross-links fibrin to fibrinolytic inhibitors, specifically α₂-antiplasmin, to increase resistance to fibrinolysis. We have previously shown that cellular FXIII (factor XIII-A [FXIII-A]), which is abundant in the platelet cytoplasm, is externalized onto the activated membrane and cross-links extracellular substrates. The contribution of cellular FXIII-A to platelet activation and platelet function has not been extensively studied. Objectives: This study aims to identify the role of platelet FXIII-A in platelet function. Methods: We used normal healthy platelets with a cell permeable FXIII inhibitor and platelets from FXIII-deficient patients as a FXIII-free platelet model in a range of platelet function and clotting tests. Results: Our data demonstrate that platelet FXIII-A enhances fibrinogen binding to the platelet surface upon agonist stimulation and improves the binding of platelets to fibrinogen and aggregation under flow in a whole-blood thrombus formation assay. In the absence of FXIII-A, platelets show reduced sensitivity to agonist stimulation, including decreased P-selectin exposure and fibrinogen binding. We show that FXIII-A is involved in platelet spreading where a lack of FXIII-A reduces the ability of platelets to fully spread on fibrinogen and collagen. Our data demonstrate that platelet FXIII-A is important for clot retraction where clots formed in its absence retracted to a lesser extent. Conclusion: Overall, this study shows that platelet FXIII-A functions during thrombus formation by aiding platelet activation and thrombus retraction in addition to its antifibrinolytic roles.

Original language	English
Article number	100200
Number of pages	15
Journal	Research and Practice in Thrombosis and Haemostasis
Volume	7
Issue number	5
Early online date	20 Jul 2023
DOIs	https://doi.org/10.1016/j.rpth.2023.100200
Publication status	Published - Jul 2023

Bibliographical note

Funding Information:
The authors thank Dr Ana Raquel Pereira from Oxford Nanoimaging for help with the direct STORM imaging and analysis. The authors also thank the microscopy core facilities at the University of Reading and the University of Aberdeen for excellent advice and use of the microscopes and facilities. The authors further thank Christopher Deane from the Oxford Haemophilia and Thrombosis centre for assistance with patient blood collection and Dr Nicholas Michael from the University of Reading Mass Spectrometry facility for performing mass spectrometry. This study was supported by the University of Reading School of Biological Sciences Seed funding grant (J.L.M.) and a grant from the British Heart Foundation (PG/16/36/31967, to C.I.J. and J.M.G.) and by the University of Aberdeen Development Trust (N.J.M. and J.L.M.). S.S. receives funding support from the Medical Research Council (MR/T024054/1). Informed consent was given by all normal health blood donors on this study with ethical approval from the University of Reading Research Ethics Committee (UREC 20/20). Informed consent was given by all patients recruited in this study with ethical approval by the Oxford Biobank (ORB Research Tissue Bank (RTB) ethics approval 09/H0606/5+5). J.L.M. conceptualized and designed the research, performed experiments, analyzed and interpreted data, and wrote the manuscript. G.L. performed the experiments and analyzed data. A.P.B. performed experiments and analyzed the data. R.S.G. performed experiments and analyzed data. A.J.U. performed experiments, analyzed data, and wrote the manuscript. N.K. performed experiments and analyzed data. T.S. performed experiments. A.S. performed experiments and analyzed data. I.S. performed experiments and analyzed data. G.B.M. performed experiments and analyzed data. R.N.B. performed experiments and analyzed data. S.S. provided patient samples and clinical advice. N.C. provided patient samples and clinical advice. J.M.G. interpreted data, obtained ethical approval for controls, and wrote the manuscript. C.S.W. interpreted data and wrote the manuscript. M.J.R.D. provided patient samples, obtained patient ethical approval, and provided clinical advice. N.J.M. designed the research, interpreted data, and wrote the manuscript. C.I.J. designed the research, interpreted data, obtained patient ethical approval, and wrote the manuscript. R.N.B is Director of Business Development at Oxford Nanoimaging.

Keywords

clot retraction
factor XIII
fibrinogen
fibrinolysis
platelet
platelet activation

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Mitchell_etal_RPTH_Platelet_Factor_XIII_VoR
© 2023 The Authors. Published by Elsevier Inc. on behalf of International Society on Thrombosis and Haemostasis. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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Cite this

Mitchell, J. L., Little, G., Bye, A. P., Gaspar, R. S., Unsworth, A. J., Kriek, N., Sage, T., Stainer, A., Sangowawa, I., Morrow, G. B., Bastos, R. N., Shapiro, S., Desborough, M. J. R., Curry, N., Gibbins, J. M., Whyte, C. S., Mutch, N. J., & Jones, C. I. (2023). Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability. Research and Practice in Thrombosis and Haemostasis, 7(5), Article 100200. https://doi.org/10.1016/j.rpth.2023.100200

Mitchell, JL, Little, G, Bye, AP, Gaspar, RS, Unsworth, AJ, Kriek, N, Sage, T, Stainer, A, Sangowawa, I, Morrow, GB, Bastos, RN, Shapiro, S, Desborough, MJR, Curry, N, Gibbins, JM, Whyte, CS , Mutch, NJ & Jones, CI 2023, 'Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability', Research and Practice in Thrombosis and Haemostasis, vol. 7, no. 5, 100200. https://doi.org/10.1016/j.rpth.2023.100200

@article{77568b10e3dd4088bd68f60ee3f2ab84,

title = "Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability",

abstract = "Background: Factor XIII (FXIII) is an important proenzyme in the hemostatic system. The plasma-derived enzyme activated FXIII cross-links fibrin fibers within thrombi to increase their mechanical strength and cross-links fibrin to fibrinolytic inhibitors, specifically α2-antiplasmin, to increase resistance to fibrinolysis. We have previously shown that cellular FXIII (factor XIII-A [FXIII-A]), which is abundant in the platelet cytoplasm, is externalized onto the activated membrane and cross-links extracellular substrates. The contribution of cellular FXIII-A to platelet activation and platelet function has not been extensively studied. Objectives: This study aims to identify the role of platelet FXIII-A in platelet function. Methods: We used normal healthy platelets with a cell permeable FXIII inhibitor and platelets from FXIII-deficient patients as a FXIII-free platelet model in a range of platelet function and clotting tests. Results: Our data demonstrate that platelet FXIII-A enhances fibrinogen binding to the platelet surface upon agonist stimulation and improves the binding of platelets to fibrinogen and aggregation under flow in a whole-blood thrombus formation assay. In the absence of FXIII-A, platelets show reduced sensitivity to agonist stimulation, including decreased P-selectin exposure and fibrinogen binding. We show that FXIII-A is involved in platelet spreading where a lack of FXIII-A reduces the ability of platelets to fully spread on fibrinogen and collagen. Our data demonstrate that platelet FXIII-A is important for clot retraction where clots formed in its absence retracted to a lesser extent. Conclusion: Overall, this study shows that platelet FXIII-A functions during thrombus formation by aiding platelet activation and thrombus retraction in addition to its antifibrinolytic roles.",

keywords = "clot retraction, factor XIII, fibrinogen, fibrinolysis, platelet, platelet activation",

author = "Mitchell, {Joanne L.} and Gemma Little and Bye, {Alexander P.} and Gaspar, {Renato S.} and Unsworth, {Amanda J.} and Neline Kriek and Tanya Sage and Alexander Stainer and Ibidayo Sangowawa and Morrow, {Gael B.} and Bastos, {Ricardo N.} and Susan Shapiro and Desborough, {Michael J.R.} and Nicola Curry and Gibbins, {Jonathan M.} and Whyte, {Claire S.} and Mutch, {Nicola J.} and Jones, {Christopher I.}",

note = "Funding Information: The authors thank Dr Ana Raquel Pereira from Oxford Nanoimaging for help with the direct STORM imaging and analysis. The authors also thank the microscopy core facilities at the University of Reading and the University of Aberdeen for excellent advice and use of the microscopes and facilities. The authors further thank Christopher Deane from the Oxford Haemophilia and Thrombosis centre for assistance with patient blood collection and Dr Nicholas Michael from the University of Reading Mass Spectrometry facility for performing mass spectrometry. This study was supported by the University of Reading School of Biological Sciences Seed funding grant (J.L.M.) and a grant from the British Heart Foundation (PG/16/36/31967, to C.I.J. and J.M.G.) and by the University of Aberdeen Development Trust (N.J.M. and J.L.M.). S.S. receives funding support from the Medical Research Council (MR/T024054/1). Informed consent was given by all normal health blood donors on this study with ethical approval from the University of Reading Research Ethics Committee (UREC 20/20). Informed consent was given by all patients recruited in this study with ethical approval by the Oxford Biobank (ORB Research Tissue Bank (RTB) ethics approval 09/H0606/5+5). J.L.M. conceptualized and designed the research, performed experiments, analyzed and interpreted data, and wrote the manuscript. G.L. performed the experiments and analyzed data. A.P.B. performed experiments and analyzed the data. R.S.G. performed experiments and analyzed data. A.J.U. performed experiments, analyzed data, and wrote the manuscript. N.K. performed experiments and analyzed data. T.S. performed experiments. A.S. performed experiments and analyzed data. I.S. performed experiments and analyzed data. G.B.M. performed experiments and analyzed data. R.N.B. performed experiments and analyzed data. S.S. provided patient samples and clinical advice. N.C. provided patient samples and clinical advice. J.M.G. interpreted data, obtained ethical approval for controls, and wrote the manuscript. C.S.W. interpreted data and wrote the manuscript. M.J.R.D. provided patient samples, obtained patient ethical approval, and provided clinical advice. N.J.M. designed the research, interpreted data, and wrote the manuscript. C.I.J. designed the research, interpreted data, obtained patient ethical approval, and wrote the manuscript. R.N.B is Director of Business Development at Oxford Nanoimaging.",

year = "2023",

month = jul,

doi = "10.1016/j.rpth.2023.100200",

language = "English",

volume = "7",

journal = "Research and Practice in Thrombosis and Haemostasis",

issn = "2475-0379",

publisher = "Wiley-Blackwell Publishing Ltd",

number = "5",

}

TY - JOUR

T1 - Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability

AU - Mitchell, Joanne L.

AU - Little, Gemma

AU - Bye, Alexander P.

AU - Gaspar, Renato S.

AU - Unsworth, Amanda J.

AU - Kriek, Neline

AU - Sage, Tanya

AU - Stainer, Alexander

AU - Sangowawa, Ibidayo

AU - Morrow, Gael B.

AU - Bastos, Ricardo N.

AU - Shapiro, Susan

AU - Desborough, Michael J.R.

AU - Curry, Nicola

AU - Gibbins, Jonathan M.

AU - Whyte, Claire S.

AU - Mutch, Nicola J.

AU - Jones, Christopher I.

N1 - Funding Information: The authors thank Dr Ana Raquel Pereira from Oxford Nanoimaging for help with the direct STORM imaging and analysis. The authors also thank the microscopy core facilities at the University of Reading and the University of Aberdeen for excellent advice and use of the microscopes and facilities. The authors further thank Christopher Deane from the Oxford Haemophilia and Thrombosis centre for assistance with patient blood collection and Dr Nicholas Michael from the University of Reading Mass Spectrometry facility for performing mass spectrometry. This study was supported by the University of Reading School of Biological Sciences Seed funding grant (J.L.M.) and a grant from the British Heart Foundation (PG/16/36/31967, to C.I.J. and J.M.G.) and by the University of Aberdeen Development Trust (N.J.M. and J.L.M.). S.S. receives funding support from the Medical Research Council (MR/T024054/1). Informed consent was given by all normal health blood donors on this study with ethical approval from the University of Reading Research Ethics Committee (UREC 20/20). Informed consent was given by all patients recruited in this study with ethical approval by the Oxford Biobank (ORB Research Tissue Bank (RTB) ethics approval 09/H0606/5+5). J.L.M. conceptualized and designed the research, performed experiments, analyzed and interpreted data, and wrote the manuscript. G.L. performed the experiments and analyzed data. A.P.B. performed experiments and analyzed the data. R.S.G. performed experiments and analyzed data. A.J.U. performed experiments, analyzed data, and wrote the manuscript. N.K. performed experiments and analyzed data. T.S. performed experiments. A.S. performed experiments and analyzed data. I.S. performed experiments and analyzed data. G.B.M. performed experiments and analyzed data. R.N.B. performed experiments and analyzed data. S.S. provided patient samples and clinical advice. N.C. provided patient samples and clinical advice. J.M.G. interpreted data, obtained ethical approval for controls, and wrote the manuscript. C.S.W. interpreted data and wrote the manuscript. M.J.R.D. provided patient samples, obtained patient ethical approval, and provided clinical advice. N.J.M. designed the research, interpreted data, and wrote the manuscript. C.I.J. designed the research, interpreted data, obtained patient ethical approval, and wrote the manuscript. R.N.B is Director of Business Development at Oxford Nanoimaging.

PY - 2023/7

Y1 - 2023/7

N2 - Background: Factor XIII (FXIII) is an important proenzyme in the hemostatic system. The plasma-derived enzyme activated FXIII cross-links fibrin fibers within thrombi to increase their mechanical strength and cross-links fibrin to fibrinolytic inhibitors, specifically α2-antiplasmin, to increase resistance to fibrinolysis. We have previously shown that cellular FXIII (factor XIII-A [FXIII-A]), which is abundant in the platelet cytoplasm, is externalized onto the activated membrane and cross-links extracellular substrates. The contribution of cellular FXIII-A to platelet activation and platelet function has not been extensively studied. Objectives: This study aims to identify the role of platelet FXIII-A in platelet function. Methods: We used normal healthy platelets with a cell permeable FXIII inhibitor and platelets from FXIII-deficient patients as a FXIII-free platelet model in a range of platelet function and clotting tests. Results: Our data demonstrate that platelet FXIII-A enhances fibrinogen binding to the platelet surface upon agonist stimulation and improves the binding of platelets to fibrinogen and aggregation under flow in a whole-blood thrombus formation assay. In the absence of FXIII-A, platelets show reduced sensitivity to agonist stimulation, including decreased P-selectin exposure and fibrinogen binding. We show that FXIII-A is involved in platelet spreading where a lack of FXIII-A reduces the ability of platelets to fully spread on fibrinogen and collagen. Our data demonstrate that platelet FXIII-A is important for clot retraction where clots formed in its absence retracted to a lesser extent. Conclusion: Overall, this study shows that platelet FXIII-A functions during thrombus formation by aiding platelet activation and thrombus retraction in addition to its antifibrinolytic roles.

AB - Background: Factor XIII (FXIII) is an important proenzyme in the hemostatic system. The plasma-derived enzyme activated FXIII cross-links fibrin fibers within thrombi to increase their mechanical strength and cross-links fibrin to fibrinolytic inhibitors, specifically α2-antiplasmin, to increase resistance to fibrinolysis. We have previously shown that cellular FXIII (factor XIII-A [FXIII-A]), which is abundant in the platelet cytoplasm, is externalized onto the activated membrane and cross-links extracellular substrates. The contribution of cellular FXIII-A to platelet activation and platelet function has not been extensively studied. Objectives: This study aims to identify the role of platelet FXIII-A in platelet function. Methods: We used normal healthy platelets with a cell permeable FXIII inhibitor and platelets from FXIII-deficient patients as a FXIII-free platelet model in a range of platelet function and clotting tests. Results: Our data demonstrate that platelet FXIII-A enhances fibrinogen binding to the platelet surface upon agonist stimulation and improves the binding of platelets to fibrinogen and aggregation under flow in a whole-blood thrombus formation assay. In the absence of FXIII-A, platelets show reduced sensitivity to agonist stimulation, including decreased P-selectin exposure and fibrinogen binding. We show that FXIII-A is involved in platelet spreading where a lack of FXIII-A reduces the ability of platelets to fully spread on fibrinogen and collagen. Our data demonstrate that platelet FXIII-A is important for clot retraction where clots formed in its absence retracted to a lesser extent. Conclusion: Overall, this study shows that platelet FXIII-A functions during thrombus formation by aiding platelet activation and thrombus retraction in addition to its antifibrinolytic roles.

KW - clot retraction

KW - factor XIII

KW - fibrinogen

KW - fibrinolysis

KW - platelet

KW - platelet activation

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U2 - 10.1016/j.rpth.2023.100200

DO - 10.1016/j.rpth.2023.100200

M3 - Article

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JO - Research and Practice in Thrombosis and Haemostasis

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M1 - 100200

ER -

Platelet factor XIII-A regulates platelet function and promotes clot retraction and stability

Abstract

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Keywords

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