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Thromb Haemost
DOI: 10.1055/a-2508-3424
Coagulation and Fibrinolysis

A Novel Fibrinolysis Resistance Capacity Assay Can Detect Fibrinolytic Phenotypes in Trauma Patients

Christopher D. Barrett    *
1   Division of Acute Care Surgery, Department of Surgery, University of Nebraska Medical Center, Omaha, Nebraska, United States
2   Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, United States
,
Yuko Suzuki    *
3   Department of Medical Physiology, Hamamatsu University School of Medicine, Hamamatsu, Japan
,
Ernest E. Moore
4   Department of Surgery, Ernest E Moore Shock Trauma Center at Denver Health, Denver, Colorado, United States
5   Department of Surgery, University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
,
Hunter B. Moore 
6   Division of Solid Organ Transplantation, Department of Surgery, AdventHealth Porter, Denver, Colorado, United States
,
Elizabeth R. Maginot
1   Division of Acute Care Surgery, Department of Surgery, University of Nebraska Medical Center, Omaha, Nebraska, United States
,
Collin M. White
2   Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, United States
,
Halima Siddiqui
1   Division of Acute Care Surgery, Department of Surgery, University of Nebraska Medical Center, Omaha, Nebraska, United States
,
Flobater I. Gawargi
2   Department of Cellular and Integrative Physiology, University of Nebraska Medical Center, Omaha, Nebraska, United States
,
James G. Chandler
5   Department of Surgery, University of Colorado Anschutz Medical Campus, Aurora, Colorado, United States
,
Angela Sauaia
7   Sauaia Statistical Solutions, LLC, Denver, Colorado, United States
,
Tetsumei Urano 
3   Department of Medical Physiology, Hamamatsu University School of Medicine, Hamamatsu, Japan
8   Department of Public Health, Shizuoka Graduate University of Public Health, Shizuoka, Japan
› Institutsangaben
Funding This study was supported in part by JSPS KAKENHI Grant Number 22H03172 (TU) and 22K08153 (YS), Japan Agency for Medical Research and Development (AMED) Grant Number 21ek0210154h0002 (TU), National Institute of General Medical Sciences grants P20-GM152326 (CDB) and T32-GM008315 (EEM, AS), and National Heart Lung and Blood Institute grant K08-HL171936 (CDB).
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Abstract

Background To evaluate residual fibrinolysis resistance activity (FRA) in plasma, a detergent-modified plasma clot lysis assay time (dPCLT) was established in which α2-antiplasmin (A2AP) and plasminogen activator inhibitor type 1 (PAI-1) are inactivated without impacting protease activity. We applied this novel assay to severely injured trauma patients’ plasma.

Material and Methods Tissue-type plasminogen activator (tPA)-induced plasma clot lysis assays were conducted after detergents- (dPCLT) or vehicle- (sPCLT) treatment, and time to 50% clot lysis was measured (“transition midpoint”, T m). Residual FRA was then calculated as ([sPCLT T m] - [dPCLT T m]/[sPCLT T m]) x100% = Δ Tm PCLT (%). Assay results were compared to rapid thromboelastography (TEG) LY30, tPA TEG LY30, and plasma fibrinolysis biomarkers in polytrauma patients’ plasma (N=43).

Results Δ Tm PCLT(%) in normal plasma (N=5) was 63.0 ± 8.3 whereas in A2AP-depleted plasma was -19.1 ± 1.3%, Plasmin-antiplasmin (PAP) complex increased after complete lysis of sPCLT, whereas that in dPCLT was negligible in normal plasma. In trauma plasma, significant correlations between Δ Tm PCLT and active PAI-1 (r = 0.85, p<0.0001), PAP complex (r = -0.85, p<0.0001), free A2AP (r = 0.66, p<0.0001), total A2AP levels (r = 0.52, p=0.001) and tPA TEG LY30 (r = -0.85, p<0.0001) were found. dPCLT in hyperfibrinolysis patients diagnosed by tPA TEG was significantly shorter than those with low fibrinolysis [10.2 ± 6.4 minutes versus 20.2 ± 2.1 minutes, p=0.0006].

Conclusion Hyperfibrinolysis after trauma is significantly related to exhaustion of FRA, and our novel assay appears to quickly assess this state and may be a useful clinical diagnostic after additional validation.

Key Points

  • We established a new clot lysis assay to measure residual fibrinolysis resistance activity after inactivating PAI-1 and A2AP by detergents without impacting protease function.

  • This novel clot lysis assay unmasked the mechanism of hyperfibrinolysis after trauma as exhaustion of fibrinolysis resistance activity, and appeared useful in quickly identifying these patients.

Keywords

fibrinolysis inhibitors - fibrinolytic disorders - surgery - hypercoagulability

Note

This paper was presented in a meeting International Society on Thrombosis and Haemostasis Annual Congress in Bangkok, Thailand, June 2024


Data Availability Statement

Supporting data can be made available to bona fide researchers upon request. Access may be requested from Dr. Christopher D. Barrett at University of Nebraska Medical Center via email at cbarrett@unmc.edu.


Authors' Contribution

C.D.B., T.U., and Y.S. prepared the manuscript with critical input, data analysis and interpretation, and revisions from all other listed authors.


Supplementary Material



Publikationsverlauf

Eingereicht: 08. Oktober 2024

Angenommen: 26. Dezember 2024

Artikel online veröffentlicht:
21. Januar 2025

© 2025. Thieme. All rights reserved.

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