Homocysteine thiolactone contributes to the prognostic value of fibrin clot structure/function in coronary artery disease

Marta Sikora, Paweł Skrzydlewski, Joanna Perła-Kaján, Hieronim Jakubowski, Marta Sikora, Paweł Skrzydlewski, Joanna Perła-Kaján, Hieronim Jakubowski

Abstract

Fibrin clot structure/function contributes to cardiovascular disease. We examined sulfur-containing metabolites as determinants of fibrin clot lysis time (CLT) and maximum absorbance (Absmax) in relation to outcomes in coronary artery disease (CAD) patients. Effects of B-vitamin/folate therapy on CLT and Absmax were studied. Plasma samples were collected from 1,952 CAD patients randomized in a 2 x 2 factorial design to (i) folic acid, vitamins B12, B6; (ii) folic acid, vitamin B12; (iii) vitamin B6; (iv) placebo for 3.8 years in the Western Norway B-Vitamin Intervention Trial. Clot lysis time (CLT) and maximum absorbance (Absmax) were determined using a validated turbidimetric assay. Acute myocardial infarction (AMI) and mortality were assessed during a 7-year follow-up. Data were analyzed using bivariate and multiple regression. Survival free of events was studied using Kaplan Mayer plots. Hazard ratios (HR) and 95% confidence intervals (CI) were estimated using Cox proportional hazards models. Baseline urinary homocysteine (uHcy)-thiolactone and plasma cysteine (Cys) were significantly associated with CLT while plasma total Hcy was significantly associated with Absmax, independently of fibrinogen, triglycerides, vitamin E, glomerular filtration rate, body mass index, age, sex plasma creatinine, CRP, HDL-C, ApoA1, and previous diseases. B-vitamins/folate did not affect CLT and Absmax. Kaplan-Meier analysis showed associations of increased baseline CLT and Absmax with worse outcomes. In Cox regression analysis, baseline CLT and Absmax (>cutoff) predicted AMI (CLT: HR 1.58, 95% CI 1.10-2.28; P = 0.013. Absmax: HR 3.22, CI 1.19-8.69; P = 0.021) and mortality (CLT: HR 2.54, 95% CI 1.40-4.63; P = 0.002. Absmax: 2.39, 95% CI 1.17-4.92; P = 0.017). After adjustments for other prognostic biomarkers these associations remained significant. Cys and uHcy-thiolactone, but not tHcy, were significant predictors of AMI in Cox regression models that included CLT. Conclusions uHcy-thiolactone and plasma Cys are novel determinants of CLT, an important predictor of adverse CAD outcomes. CLT and Absmax were not affected by B-vitamin/folate therapy, which could account for the lack of efficacy of such therapy in CAD. Trial registration: URL: https://ichgcp.net/clinical-trials-registry/NCT00354081" title="See in ClinicalTrials.gov">NCT00354081.

Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1. Kaplan-Meier analysis of outcome events…
Fig 1. Kaplan-Meier analysis of outcome events according to CLT and Absmax cutoffs.
A. Survival free of AMI in CLT group 0 (CLT ≤ 397.5 s) and group 1 (CLT > 397.5) vs. time (days). B. Survival without mortality in CLT group 0 (CLT ≤ 532.5 s) and group 1 (CLT > 532.5 s). C. Survival free of AMI in Absmax group 0 (Absmax ≤ 0.025) and group 1 (Amax > 0.025) vs. time (days). D. Survival without mortality in Absmax group 0 (Absmax ≤ 0.169) and group 1 (Absmax > 0.169).

References

    1. Undas A, Ariens RA. Fibrin clot structure and function: a role in the pathophysiology of arterial and venous thromboembolic diseases. Arteriosclerosis, thrombosis, and vascular biology 2011; 31: e88–99. doi: 10.1161/ATVBAHA.111.230631
    1. Carter AM, Cymbalista CM, Spector TD, Grant PJ, Euro CI. Heritability of clot formation, morphology, and lysis: the EuroCLOT study. Arteriosclerosis, thrombosis, and vascular biology 2007; 27: 2783–9. doi: 10.1161/ATVBAHA.107.153221
    1. Mudd SH, Finkelstein JD, Refsum H, et al.. Homocysteine and its disulfide derivatives: a suggested consensus terminology. Arteriosclerosis, thrombosis, and vascular biology 2000; 20: 1704–6. doi: 10.1161/01.atv.20.7.1704
    1. Refsum H, Smith AD, Ueland PM, et al. Facts and recommendations about total homocysteine determinations: an expert opinion. Clinical chemistry 2004; 50: 3–32. doi: 10.1373/clinchem.2003.021634
    1. Jakubowski H. Homocysteine Modification in Protein Structure/Function and Human Disease. Physiological Reviews 2019; 99: 555–604. doi: 10.1152/physrev.00003.2018
    1. Borowczyk K, Piechocka J, Glowacki R, et al. Urinary excretion of homocysteine thiolactone and the risk of acute myocardial infarction in coronary artery disease patients: the WENBIT trial. J Intern Med 2019; 285: 232–44. doi: 10.1111/joim.12834
    1. Undas A, Perla J, Lacinski M, Trzeciak W, Kazmierski R, Jakubowski H. Autoantibodies against N-homocysteinylated proteins in humans: implications for atherosclerosis. Stroke 2004; 35: 1299–304. doi: 10.1161/01.STR.0000128412.59768.6e
    1. Chwatko G, Jakubowski H. Urinary excretion of homocysteine-thiolactone in humans. Clinical chemistry 2005; 51: 408–15. doi: 10.1373/clinchem.2004.042531
    1. Chwatko G, Jakubowski H. The determination of homocysteine-thiolactone in human plasma. Analytical biochemistry 2005; 337: 271–7. doi: 10.1016/j.ab.2004.11.035
    1. Jakubowski H. Quantification of urinary S- and N-homocysteinylated protein and homocysteine-thiolactone in mice. Analytical biochemistry 2016; 508: 118–23. doi: 10.1016/j.ab.2016.06.002
    1. Ebbing M, Bleie O, Ueland PM, et al. Mortality and cardiovascular events in patients treated with homocysteine-lowering B vitamins after coronary angiography: a randomized controlled trial. Jama 2008; 300: 795–804. doi: 10.1001/jama.300.7.795
    1. Sulo E, Vollset SE, Nygard O, et al. Trends in 28-day and 1-year mortality rates in patients hospitalized for a first acute myocardial infarction in Norway during 2001–2009: a "Cardiovascular disease in Norway" (CVDNOR) project. J Intern Med 2015; 277: 353–61. doi: 10.1111/joim.12266
    1. Winther-Larsen A, Christiansen MK, Larsen SB, et al. The ABO Locus is Associated with Increased Fibrin Network Formation in Patients with Stable Coronary Artery Disease. Thrombosis and haemostasis 2020; 120: 1248–56. doi: 10.1055/s-0040-1713753
    1. Sumaya W, Wallentin L, James SK, et al. Impaired Fibrinolysis Predicts Adverse Outcome in Acute Coronary Syndrome Patients with Diabetes: A PLATO Sub-Study. Thrombosis and haemostasis 2020; 120: 412–22. doi: 10.1055/s-0039-1701011
    1. Moroz LA, Gilmore NJ. Inhibition of plasmin-mediated fibrinolysis by vitamin E. Nature 1976; 259: 235–7. doi: 10.1038/259235a0
    1. Zabczyk M, Hondo L, Krzek M, Undas A. High-density cholesterol and apolipoprotein AI as modifiers of plasma fibrin clot properties in apparently healthy individuals. Blood Coagul Fibrinolysis 2013; 24: 50–4. doi: 10.1097/MBC.0b013e32835a083c
    1. Zabczyk M, Natorska J, Undas A. Fibrin Clot Properties in Atherosclerotic Vascular Disease: From Pathophysiology to Clinical Outcomes. J Clin Med 2021; 10. doi: 10.3390/jcm10132999
    1. Swanepoel AC, de Lange-Loots Z, Cockeran M, Pieters M. Lifestyle Influences Changes in Fibrin Clot Properties Over a 10-Year Period on a Population Level. Thrombosis and haemostasis 2022; 122: 67–79. doi: 10.1055/a-1492-6143
    1. Neergaard-Petersen S, Hvas AM, Kristensen SD, et al. The influence of type 2 diabetes on fibrin clot properties in patients with coronary artery disease. Thrombosis and haemostasis 2014; 112: 1142–50. doi: 10.1160/TH14-05-0468
    1. Siudut J, Iwaniec T, Plens K, Pieters M, Undas A. Determinants of plasma fibrin clot lysis measured using three different assays in healthy subjects. Thromb Res 2021; 197: 1–7. doi: 10.1016/j.thromres.2020.10.014
    1. Meltzer ME, Lisman T, de Groot PG, Meijers JC, le Cessie S, Doggen CJ, et al.. Venous thrombosis risk associated with plasma hypofibrinolysis is explained by elevated plasma levels of TAFI and PAI-1. Blood 2010; 116: 113–21. doi: 10.1182/blood-2010-02-267740
    1. Undas A, Brozek J, Jankowski M, Siudak Z, Szczeklik A, Jakubowski H. Plasma homocysteine affects fibrin clot permeability and resistance to lysis in human subjects. Arteriosclerosis, thrombosis, and vascular biology 2006; 26: 1397–404. doi: 10.1161/01.ATV.0000219688.43572.75
    1. Undas A, Plicner D, Stepien E, Drwila R, Sadowski J. Altered fibrin clot structure in patients with advanced coronary artery disease: a role of C-reactive protein, lipoprotein(a) and homocysteine. J Thromb Haemost 2007; 5: 1988–90. doi: 10.1111/j.1538-7836.2007.02637.x
    1. Undas A, Stepien E, Tracz W, Szczeklik A. Lipoprotein(a) as a modifier of fibrin clot permeability and susceptibility to lysis. J Thromb Haemost 2006; 4: 973–5. doi: 10.1111/j.1538-7836.2006.01903.x
    1. Krzek M, Ciesla-Dul M, Zabczyk M, Undas A. Fibrin clot properties in women heterozygous for factor V Leiden mutation: effects of oral contraceptives. Thromb Res 2012; 130: e216–21. doi: 10.1016/j.thromres.2012.08.302
    1. Jakubowski H, Boers GH, Strauss KA. Mutations in cystathionine beta-synthase or methylenetetrahydrofolate reductase gene increase N-homocysteinylated protein levels in humans. FASEB journal: official publication of the Federation of American Societies for Experimental Biology 2008; 22: 4071–6.
    1. Jakubowski H. Homocysteine is a protein amino acid in humans. Implications for homocysteine-linked disease. The Journal of biological chemistry 2002; 277: 30425–8. doi: 10.1074/jbc.C200267200
    1. Sauls DL, Lockhart E, Warren ME, Lenkowski A, Wilhelm SE, Hoffman M. Modification of fibrinogen by homocysteine thiolactone increases resistance to fibrinolysis: a potential mechanism of the thrombotic tendency in hyperhomocysteinemia. Biochemistry 2006; 45: 2480–7. doi: 10.1021/bi052076j
    1. Glowacki R, Jakubowski H. Cross-talk between Cys34 and lysine residues in human serum albumin revealed by N-homocysteinylation. The Journal of biological chemistry 2004; 279: 10864–71. doi: 10.1074/jbc.M313268200
    1. Leander K, Blomback M, Wallen H, He S. Impaired fibrinolytic capacity and increased fibrin formation associate with myocardial infarction. Thrombosis and haemostasis 2012; 107: 1092–9. doi: 10.1160/TH11-11-0760
    1. Saraf S, Christopoulos C, Salha IB, Stott DJ, Gorog DA. Impaired endogenous thrombolysis in acute coronary syndrome patients predicts cardiovascular death and nonfatal myocardial infarction. J Am Coll Cardiol 2010; 55: 2107–15. doi: 10.1016/j.jacc.2010.01.033
    1. Sumaya W, Wallentin L, James SK, et al. Fibrin clot properties independently predict adverse clinical outcome following acute coronary syndrome: a PLATO substudy. European heart journal 2018; 39: 1078–85. doi: 10.1093/eurheartj/ehy013
    1. Neergaard-Petersen S, Larsen SB, Grove EL, Kristensen SD, Ajjan RA, Hvas AM. Imbalance between Fibrin Clot Formation and Fibrinolysis Predicts Cardiovascular Events in Patients with Stable Coronary Artery Disease. Thrombosis and haemostasis 2020; 120: 75–82. doi: 10.1055/s-0039-1700873
    1. Undas A, Stepien E, Glowacki R, Tisonczyk J, Tracz W, Jakubowski H. Folic acid administration and antibodies against homocysteinylated proteins in subjects with hyperhomocysteinemia. Thrombosis and haemostasis 2006; 96: 342–7. doi: 10.1160/TH06-04-0228
    1. Bleie O, Semb AG, Grundt H, et al. Homocysteine-lowering therapy does not affect inflammatory markers of atherosclerosis in patients with stable coronary artery disease. J Intern Med 2007; 262: 244–53. doi: 10.1111/j.1365-2796.2007.01810.x
    1. Maron BA, Loscalzo J. The treatment of hyperhomocysteinemia. Annu Rev Med 2009; 60: 39–54. doi: 10.1146/annurev.med.60.041807.123308
    1. Collet JP, Allali Y, Lesty C, et al. Altered fibrin architecture is associated with hypofibrinolysis and premature coronary atherothrombosis. Arteriosclerosis, thrombosis, and vascular biology 2006; 26: 2567–73. doi: 10.1161/01.ATV.0000241589.52950.4c

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