Tissue Factor Pathway–Driven Initial Thrombin Generation is Associated with Hypercoagulability in Obesity

Abstract

Background and Objectives

Initial thrombin (FIIa) generated via the tissue factor (TF) pathway plays a crucial role in amplifying coagulation. There is growing evidence that the TF pathway might contribute to hypercoagulation in obesity. However, it is unclear if the initial generation of FIIa (TG) is associated with hypercoagulation in obesity due to the lack of appropriate assays. This study aims to evaluate association between TF pathway-driven initial TG and hypercoagulability in obesity.

Methods

We measured the initial TG levels in plasma from male Tsumura Suzuki obese diabetes (TSOD) mice and overweight subjects using the highly sensitive TG assay. To induce initial TG, TF was added to the plasma and incubated at 37°C for up to 3 minutes. After quenching the TG, we quantified the generated FIIa by kinetically monitoring its amidolytic activity with a fluorogenic substrate.

Results and Conclusion

We observed that initial TG levels were significantly higher in TSOD mice (n = 31) compared with non-obese mice (n = 32). Even in the absence of exogenous TF, initial TG levels in obese mice and overweight individuals were elevated when procoagulant phospholipids were added alone. Moreover, the increased initial TG that the inhibitory anti-TF antibody abolished was detectable in reconstituted plasma including pellets prepared by high-speed centrifugation of plasma from obese mice, not in plasma supernatant. We attributed the promotion of the initial TG to the increase in procoagulant TF-bearing microvesicles in circulation. Based on the findings, measuring TF pathway-driven initial TG could be a valuable method for assessing hypercoagulability in obesity.

Keywords

obesity - tissue factor - initial thrombin generation - hypercoagulability - thromboembolic diseases

Authors' Contribution

Contribution: Y. K. conceived and planned the study, performed assays, interpreted results, and wrote the manuscript; R.I. performed assays; S.N., K.Y., and E.M. performed the clinical study with healthy volunteers and reviewed the manuscript; K.I. performed the clinical study with overweight subjects; R.M.-K. and N.O. performed the study with obese mice and reviewed the manuscript; F.S. reviewed the manuscript.

Publikationsverlauf

Eingereicht: 17. September 2024

Angenommen: 05. März 2025

Accepted Manuscript online:
06. März 2025

Artikel online veröffentlicht:
31. März 2025

• References

• 1 Kahn SE, Hull RL, Utzschneider KM. Mechanisms linking obesity to insulin resistance and type 2 diabetes. Nature 2006; 444 (7121) 840-846
• 2 Coutinho T, Goel K, Corrêa de Sá D. et al. Central obesity and survival in subjects with coronary artery disease: a systematic review of the literature and collaborative analysis with individual subject data. J Am Coll Cardiol 2011; 57 (19) 1877-1886
• 3 Stein PD, Beemath A, Olson RE. Obesity as a risk factor in venous thromboembolism. Am J Med 2005; 118 (09) 978-980
• 4 Samad F, Ruf W. Inflammation, obesity, and thrombosis. Blood 2013; 122 (20) 3415-3422
• 5 De Pergola G, Pannacciulli N. Coagulation and fibrinolysis abnormalities in obesity. J Endocrinol Invest 2002; 25 (10) 899-904
• 6 Kario K, Matsuo T, Kobayashi H, Matsuo M, Sakata T, Miyata T. Activation of tissue factor-induced coagulation and endothelial cell dysfunction in non-insulin-dependent diabetic patients with microalbuminuria. Arterioscler Thromb Vasc Biol 1995; 15 (08) 1114-1120
• 7 Guagnano MT, Romano M, Falco A. et al. Leptin increase is associated with markers of the hemostatic system in obese healthy women. J Thromb Haemost 2003; 1 (11) 2330-2334
• 8 Ohkura N, Shirakura M, Nakatani E, Oishi K, Atsumi G. Associations between plasma PAI-1 concentrations and its expressions in various organs in obese model mice. Thromb Res 2012; 130 (06) e301-e304
• 9 Mann KG. Thrombin generation in hemorrhage control and vascular occlusion. Circulation 2011; 124 (02) 225-235
• 10 Hemker HC, Giesen P, Al Dieri R. et al. Calibrated automated thrombin generation measurement in clotting plasma. Pathophysiol Haemost Thromb 2003; 33 (01) 4-15
• 11 Tripodi A. Thrombin generation assay and its application in the clinical laboratory. Clin Chem 2016; 62 (05) 699-707
• 12 Ohkura N, Enjyoji K, Kamikubo Y, Kato H. A novel degradation pathway of tissue factor pathway inhibitor: incorporation into fibrin clot and degradation by thrombin. Blood 1997; 90 (05) 1883-1892
• 13 Ohkura N, Tamura K, Tanaka A, Matsuda J, Atsumi G. Experimental study on the hemostatc activity of Pollen Typhae: a traditional folk medicine used by external and oral application. Blood Coagul Fibrinolysis 2011; 22 (08) 631-636
• 14 Obesity: preventing and managing the global epidemic. Report of a WHO Consultation. World Health Organ Tech Rep Ser 2000; 894: i-xii , 1–253
• 15 Kamikubo Y, Mendolicchio GL, Zampolli A. et al. Selective factor VIII activation by the tissue factor-factor VIIa-factor Xa complex. Blood 2017; 130 (14) 1661-1670
• 16 de Laat-Kremers R, Di Castelnuovo A, van der Vorm L. et al; Moli-sani Investigators. Increased BMI and blood lipids are associated with a hypercoagulable state in the Moli-sani cohort. Front Cardiovasc Med 2022; 9: 897733
• 17 National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) final report. Circulation 2002; 106 (25) 3143-3421
• 18 Suzuki W, Iizuka S, Tabuchi M. et al. A new mouse model of spontaneous diabetes derived from ddY strain. Exp Anim 1999; 48 (03) 181-189
• 19 Tsuneyama K, Nishitsuji K, Matsumoto M. et al. Animal models for analyzing metabolic syndrome-associated liver diseases. Pathol Int 2017; 67 (11) 539-546
• 20 Yusuf S, Hawken S, Ounpuu S. et al; INTERHEART Study Investigators. Obesity and the risk of myocardial infarction in 27,000 participants from 52 countries: a case-control study. Lancet 2005; 366 (9497) 1640-1649
• 21 Campello E, Zabeo E, Radu CM. et al. Hypercoagulability in overweight and obese subjects who are asymptomatic for thrombotic events. Thromb Haemost 2015; 113 (01) 85-96
• 22 Cimenti C, Mangge H, Haidl H, Zach D, Muntean W. Thrombin generation in severely obese children. J Thromb Haemost 2006; 4 (08) 1834-1836
• 23 Sonnevi K, Tchaikovski SN, Holmström M. et al. Obesity and thrombin-generation profiles in women with venous thromboembolism. Blood Coagul Fibrinolysis 2013; 24 (05) 547-553
• 24 Ay L, Kopp HP, Brix JM. et al. Thrombin generation in morbid obesity: significant reduction after weight loss. J Thromb Haemost 2010; 8 (04) 759-765
• 25 Butenas S, van 't Veer C, Mann KG. Evaluation of the initiation phase of blood coagulation using ultrasensitive assays for serine proteases. J Biol Chem 1997; 272 (34) 21527-21533
• 26 Mann KG, Orfeo T, Butenas S, Undas A, Brummel-Ziedins K. Blood coagulation dynamics in haemostasis. Hamostaseologie 2009; 29 (01) 7-16
• 27 Diamant M, Nieuwland R, Pablo RF, Sturk A, Smit JWA, Radder JK. Elevated numbers of tissue-factor exposing microparticles correlate with components of the metabolic syndrome in uncomplicated type 2 diabetes mellitus. Circulation 2002; 106 (19) 2442-2447
• 28 Wang J, Ciaraldi TP, Samad F. Tissue factor expression in obese type 2 diabetic subjects and its regulation by antidiabetic agents. J Obes 2015; 2015: 291209
• 29 Ay L, Thaler J, Brix JM. et al. Decrease in microvesicle-associated tissue factor activity in morbidly obese patients after bariatric surgery. Int J Obes (Lond) 2016; 40 (05) 768-772
• 30 Ayer JG, Song C, Steinbeck K, Celermajer DS, Ben Freedman S. Increased tissue factor activity in monocytes from obese young adults. Clin Exp Pharmacol Physiol 2010; 37 (11) 1049-1054
• 31 Kopp CW, Kopp HP, Steiner S. et al. Weight loss reduces tissue factor in morbidly obese patients. Obes Res 2003; 11 (08) 950-956
• 32 Wood JP, Ellery PER, Maroney SA, Mast AE. Biology of tissue factor pathway inhibitor. Blood 2014; 123 (19) 2934-2943