A novel method to quantify fibrin–fibrin and fibrin–α2-antiplasmin cross-links in thrombi formed from human trauma patient plasma.

Morrow, Gael B.; Flannery, Sarah; Charles, Philip D.; Heilig, Raphael; Feller, Timea; McQuilten, Zoe; Wake, Elizabeth; Ariens, Robert A.S.; Winearls, James; Mutch, Nicola J.; Fischer, Roman; Laffan, Mike A.; Curry, Nicola

Abstract

The widespread use of the anti-fibrinolytic agent, tranexamic acid (TXA), interferes with the quantification of fibrinolysis by dynamic laboratory assays such as clot lysis, making it difficult to measure fibrinolysis in many trauma patients. At the final stage of coagulation, Factor XIIIa (FXIIIa) catalyses the formation of fibrin-fibrin and fibrin-α2-antiplasmin (α2AP) cross-links which increases clot mechanical strength and resistance to fibrinolysis. Here, we develop a method to quantify fibrin-fibrin and fibrin-α2AP cross-links that avoids the challenges posed by TXA in determining fibrinolytic resistance in conventional assays. Fibrinogen alpha chain (FGA-FGA), fibrinogen gamma chain (FGG-FGG) and FGA-α2AP cross-links were quantified using liquid-chromatography-mass spectrometry (LC-MS) and parallel reaction monitoring (PRM) in paired plasma samples from trauma patients pre- and post-fibrinogen replacement. Differences in the abundance of cross-links in trauma patients receiving cryoprecipitate (cryo) or fibrinogen concentrate (Fg-C) were analysed. The study found that the abundance of cross-links was significantly increased in trauma patients post-cryo, but not Fg-C, transfusion (p < 0.0001). The abundance of cross-links was positively correlated with the toughness of individual fibrin fibres, the peak thrombin concentration and FXIII antigen (p < 0.05). We have developed a novel method that allows us to quantify fibrin cross-links in trauma patients who have received TXA, providing an indirect measure of fibrinolytic resistance. Using this novel approach we have avoided the effect of TXA and shown that cryo increases fibrin-fibrin and fibrin-α2AP cross-linking when compared to Fg-C, highlighting the importance of FXIII in clot formation and stability in trauma patients.

Citation

MORROW, G.B., FLANNERY, S., CHARLES, P.D., HEILIG, R., FELLER, T., MCQUILTEN, Z., WAKE, E., ARIENS, R.A.S., WINEARLS, J., MUTCH, N.J., FISCHER, R., LAFFAN, M.A. and CURRY, N. 2024. A novel method to quantify fibrin–fibrin and fibrin–α2-antiplasmin cross-links in thrombi formed from human trauma patient plasma. Journal of thrombosis and haemostasis [online], 22(6), pages 1758-1771. Available from: https://doi.org/10.1016/j.jtha.2024.03.001

• Journal Article Type: Article
• Acceptance Date: Mar 4, 2024
• Online Publication Date: Mar 9, 2024
• Publication Date: Jun 30, 2024
• Deposit Date: Mar 11, 2024
• Publicly Available Date: Mar 11, 2024
• Journal: Journal of thrombosis and haemostasis
• Print ISSN: 1538-7933
• Electronic ISSN: 1538-7836
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 22
• Issue: 6
• Pages: 1758-1771
• DOI: https://doi.org/10.1016/j.jtha.2024.03.001
• Keywords: Fibrinogen; Fibrinogen cross-linking; Fibrinolysis; Anti-fibrinolytic agents; Trauma coagulopathy; Tranexamic acid
• Public URL: https://rgu-repository.worktribe.com/output/2270247
• Additional Information: This article has been published with separate supporting information. This supporting information has been incorporated into a single file on this repository and can be found at the end of the file associated with this output.