Skip to main content
NCBI home page
NIHPA Author Manuscripts logoLink to NIHPA Author Manuscripts
. Author manuscript; available in PMC: 2013 Apr 1.
Published in final edited form as: J Thromb Haemost. 2012 Apr;10(4):719–722. doi: 10.1111/j.1538-7836.2012.04665.x

Genetic variation in F3 (tissue factor) and the risk of incident venous thrombosis: meta-analysis of 8 studies

Nicholas L Smith 1, John A Heit 1, Weihong Tang 1, Martina Teichert 1, Daniel I Chasman 1, Pierre-Emmanuel Morange 1, on behalf of the Venous Thrombosis Genetic Replication Consortium
PMCID: PMC3397243  NIHMSID: NIHMS358658  PMID: 22340074

The coagulation factor III gene (F3) produces tissue factor (TF), a cell-surface glycoprotein that is expressed throughout the body including inside the vascular wall; it also circulates in microparticles.1 Tissue factor and its activated cofactor, coagulation factor VII (FVIIa), initiate the human extrinsic coagulation cascade.2 The TF-FVIIa complex is also involved in other biologic processes such as inflammation, angiogenesis, and tumor growth.3, 4 The F3 locus was 1 of 3 associated with plasma levels of fibrin fragment D-dimer in a recent genome-wide association study.5 The F3 rs12029080 was associated with the smallest p-value (6.4×10−22) at this locus and is located 46.0 kb upstream from the start of transcription. Each copy of the G allele was associated with a 10.4% (genome-wide confidence interval [CI]: 6.6-14.3%) increase in median D-dimer level. The F3 association was novel and suggests that TF has a role in coagulation propagation and possibly thrombotic risk beyond its well characterized role in coagulation initiation. We hypothesize that the G allele of the F3 rs12029080 variant, which was associated with elevated plasma D-dimer levels, would also be associated with an increased risk of incident VT.

A meta-analysis of 9 datasets from 8 epidemiologic studies of incident VT was performed. The 8 studies included the Heart and Vascular Health Study, 6 the Longitudinal Investigation of Thromboembolism Etiology (LITE),7 the Marseille Thrombosis Association (MARTHA) Study,8 a second French case-control study on early-onset VT (EOVT),9 the Mayo VTE Study,10 the Rotterdam Study,11 and the Women’s Genome Health Study.12 The LITE study included data from 2 large cohort studies, the Cardiovascular Health Study and the Atherosclerosis Risk in Communities Study.13, 14 Each study used regression methods to characterize the association between rs12029080 and incident VT in European-ancestry participants using an additive genetic model that adjusted for age, sex, and study-design variables. The resultant coefficients and standard errors from the 9 association-studies were meta-analyzed using fixed-effect inverse-variance weights. A 1-sided level of 0.05 determined statistical significance. Given the positive association between the rs12029080 G-allele and D-dimer levels, a negative beta-coefficient between the G-allele and VT risk (representing a decreased risk of VT) would not be considered a significant finding.

Characteristics of the studies and populations are provided in the Table. There were 5117 incident VT events across the studies, 1809 (35%) were for a PE, with or without DVT, and 2731 (53%) were diagnosed in the absence of a recent hospitalization (90 days before) or cancer (within 5 years). The average age of the subject at the time of VT onset was 54.1 years and 34% of the events were among men. (The HVH and WGHS were exclusively women.) Results are also provided in the Table. The average minor allele frequency of the rs12029080 G allele was 0.299 (range 0.286-0.317). The inverse-variance weighted beta-coefficient was 0.026 (range −0.108 to 0.199) with a standard error of 0.029 (range 0.060 to 0.353). Expressed as an odds ratio with 1-sided 95% confidence intervals, the coefficient for the G-allele was not associated with a significantly increased risk of incident VT: 1.03 (95% CI: 0.98-1.08); p-value = 0.190. The Q-statistic for the heterogeneity test across studies was 10.6 and yielded a p-value of 0.22. In 2 of the larger studies (HVH and MARTHA), we estimated the risk in idiopathic cases and found no meaningful difference in risk estimates or consistency of the risk estimates for idiopathic cases.

Table 1.

Associations between F3 rs12029080 G-allele and incident venous thrombosis

Design Events,
n		 Male,
%		 Age*,
yrs		 PE,
%		 Idiopathic,
%		 MAF Beta coefficient
(SE)		 Odd ratio (95% CI)
HVH CC 656 0 64.7 48 59 0.304 −0.073 (0.092) 0.93 (0.8, 1.08)
EOVT CC 419 55 36.0 0 100 0.327 0.091 (0.089) 1.10 (0.95, 1.27)
LITE (ARIC) Cohort 248 47 56.0 42 36 0.299 0.040 (0.097) 1.04 (0.89, 1.22)
LITE (CHS) Cohort 119 43 73.0 30 39 0.317 0.199 (0.138) 1.22 (0.97, 1.53)
MARTHA CC 1542 34 47.0 22 55 0.316 0.023 (0.060) 1.02 (0.93, 1.13)
Mayo VTE CC 1503 50 54.9 50 43 0.299 −0.068 (0.063) 0.93 (0.84, 1.04)
RS-1 Cohort 177 43 69.5 44 53 0.291 −0.108 (0.120) 0.90 (0.74, 1.09)
RS-2 Cohort 19 63 66.3 42 53 0.286 0.161 (0.353) 1.17 (0.66, 2.10)
WGHS Cohort 434 0 65.4 41 44 0.299 0.173 (0.076) 1.19 (1.05, 1.35)
Combined 5117 34 54.1 35 53 0.299** 0.026 (0.029)*** 1.03 (0.89-1.08)
Open in a new tab

CI = confidence interval; MAF = minor allele frequency; SE = standard error. The Rotterdam Study analyzed 2 subcohorts separately: RS-1 and RS-2.

*

Age at the time of the event among case subjects.

**

Averaged.

***

Weighted by inverse of variance.

Our inability to find strong evidence for an association between the G allele of F3 rs12029080 and an increased risk of incident VT stands apart from our findings when we investigated other genetic loci that predicted plasma D-dimer levels, factor V (F5) and fibrinogen alpha chain (FGA).5 The F5 Leiden R506Q (rs6025) variant accounted for the signal at the F5 locus and was associated with a 24% (genome-wide CI: 1.3-52%) difference in median D-dimer level in a subset of the cohorts where this variant was measured. The FGA T331A (rs6050) variant, also known as T312A, accounted for the signal at the FGA locus and was associated with a 6.3% (genome-wide CI: 2.3-10.4%) difference. The F5 and FGA D-dimer associations had been previously reported and both the rs6025 and rs6050 variants are associated with incident venous thrombosis (VT).15-20

The D-dimer measure quantifies by-products of fibrin clot lysis and is considered a global measure of coagulation. The F5 Leiden rs6025 variant increases D-dimer levels by inducing resistance to activated protein C thus inhibiting anti-coagulation, promoting fibrin production, and increasing the risk of VT. The FGA rs6050 variant is not associated with increased fibrinogen production rather it enhances factor XIIIa cross-linking in fibrin clots.21, 22 The enhanced crosslinking increases the risk of VT but its relation to D-dimer production is not known.

Our hypothesis that the increased D-dimer levels associated with the F3 rs1209080 variant would also be associated with increased risk of VT was not well supported by our data. There are several possible explanations for this, of which the most likely is that the pathway leading to increased D-dimer production is independent of any pathway leading to an increase in the risk of thrombosis. The F3-D-dimer pathway may be more related to fibrinolysis or to constitutional properties of the fibrin clot than to coagulation propensity. It is also possible that the sample-size of this meta-analysis was too limited to detect a small association between F3 rs12029080 and risk of incident VT. Nonetheless, the upper confidence limit of the odds ratio was 1.08 which suggests that any association would likely be modest and less than an 8% increase in the relative risk. The lack of statistical significance for the heterogeneity test across studies suggests that random variation is a more likely explanation of the range of odds ratios than any true underlying difference in relative risk.

In summary, we did not find that the F3 rs12029080 variant, which has previously been shown to be associated with D-dimer levels, was also associated with risk of incident VT in participants of European ancestry.

Acknowledgments

All studies would like to acknowledge the many tens of thousands of individuals who participated in these research projects. In addition, the Mayo VTE study would like to acknowledge Julie M. Cunningham, Ph.D., Yan W. Asmann, Ph.D., Sebastian Armasu, M.S., Martha E Matsumoto, Elysia Jeavons, Cynthia E. Regnier, R.N., Jennifer L. Alkhamis, Lonnie M. Heimer, Rene M. Weatherly, Roger A. Mueller, David Tines, Ailing Xue, M.D., Ross A. Miller, Joseph J. Larson, and Ann F. Beauseigneur for their excellent assistance.

Funding: The Heart and Vascular Health study is supported by the National Health Lung and Blood Institute grants HL43201, HL60739, HL68986, HL73410, HL74745, HL85251, and HL95080, and by a grant from the Leducq Foundation, Paris, France for the development of Transatlantic Networks of Excellence in Cardiovascular Research.

The Longitudinal Investigation of Thromboembolism Etiology was funded by National Heart, Lung, and Blood Institute grant R01 HL59367. Within LITE, the ARIC study was funded by NHLBI contracts N01-HC-55015, N01-HC-55016, N01-HC-55018, N01-HC-55019, N01-HC-55020, N01-HC-55021, and N01-HC-55022 and the CHS research was supported by NHLBI contracts N01-HC-85239, N01-HC-85079 through N01-HC-85086; N01-HC-35129, N01 HC-15103, N01 HC-55222, N01-HC-75150, N01-HC-45133 and NHLBI grants HL080295, HL075366, HL087652, HL105756 with additional contribution from NINDS. Additional support was provided through AG-023629, AG-15928, AG-20098, and AG-027058 from the NIA. DNA handling and genotyping was supported in part by National Center for Research Resources CTSI grant UL 1RR033176 and National Institute of Diabetes and Digestive and Kidney Diseases grant DK063491 to the Southern California Diabetes Endocrinology Research Center.

The Rotterdam Study is supported by the Erasmus Medical Center and Erasmus University Rotterdam, The Netherlands Organization for Scientific Research (NWO), The Netherlands Organization for Health Research and Development (ZonMw), the Research Institute for Diseases in the Elderly (RIDE), The Netherlands Genomics Initiative, the Ministry of Education, Culture and Science, the Ministry of Health, Welfare and Sports, the European Commission (DG XII), and the Municipality of Rotterdam.

The EOVT and MARTHA projects were supported by a grant from the Program Hospitalier de Recherche Clinique. The 3C Study, that served as control group for the MARTHA project, was conducted under a partnership agreement between Inserm, the Victor Segalen –Bordeaux II University and Sanofi-Synthélabo. The Fondation pour la Recherche Médicale funded the preparation and first phase of the study. The 3C-Study is also supported by the Caisse Nationale Maladie des Travailleurs Salariés, Direction Générale de la Santé, Mutuelle Générale de l’Education Nationale, the Institut de la Longévité, Agence Française de Sécurité Sanitaire des Produits de Santé, the Regional Governments of Aquitaine, Bourgogne and Languedoc-Roussillon and, the Fondation de France, the Ministry of Research-Inserm Programme ’Cohorts and collection of biological material’. The Lille Génopôle received an unconditional grant from Eisai.

The Mayo VTE Study is funded, in part, by grants from the National Institutes of Health, National Heart, Lung and Blood Institute (HL83141) and National Human Genome Research Institute (HG04735), U.S. Public Health Service, and by the Mayo Foundation.

The Women’s Genome Health Study is supported by HL043851 and HL69757 from the National Heart, Lung, and Blood Institute and CA 047988 from the National Cancer Institute, the Donald W. Reynolds Foundation and the Fondation Leducq, with collaborative scientific support and funding for genotyping provided by Amgen

Footnotes

Collaborators: Barbara McKnight, PhD, Nathan Pankratz, PhD, Frank W. G. Leebeek, MD, PhD, Lynda M. Rose, MS, Philippe Amouyel, MD, PhD, Andre G. Uitterlinden, PhD, Albert Hofman, MD, PhD, Mark Lathrop, PhD, Jean-Charles Lambert, PhD, Mary Cushman, MD, MSc, Mariza de Andrade, PhD, Aaron R. Folsom, MD, Bruno H. Stricker, MB, PhD, Paul M Ridker, MD, MPH, David-Alexandre Tregouet, PhD

Affiliations: Department of Biostatistics (BM), University of Washington, Seattle WA USA 98101; Departments of Pathology (MC) and Medicine (MC), University of Vermont, Burlington, VT USA 05405; Division of Epidemiology (ARF) and Department of Laboratory Medicine and Pathology (NP), University of Minnesota, Minneapolis MN 55455; INSERM U744 (PA), F-59019 Lille, France; Institut Pasteur de Lille, F-59019 Lille, France Université de Lille Nord de France, F-59019 Lille, France; Commissariat à l’Energie Atomique (ML), Institut de Génomique, Centre National de Génotypage, Evry, France ; INSERM U744 (JCL), F-59019 Lille, France; Institut Pasteur de Lille, F-59019 Lille, France Université de Lille Nord de France, F-59019 Lille, France; TBD; INSERM UMR_S 937 (DAT), ICAN Institute, Université Pierre et Marie Curie, Paris 6; F-75013, Paris, France; Division of Biomedical Statistics and Informatics (MdA), Department of Health Sciences Research, Mayo Clinic, Rochester, MN USA 55905; Department of Epidemiology (AGU, AF, BHS), Internal Medicine (AGU), and Haematology (FWGL), Erasmus Medical Center, 3000 CA Rotterdam, The Netherlands; Division of Preventive Medicine (LMR, PMR), Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA, 02215.

Disclosures: There are no potential conflicts of interest for any author.

References

  • 1.Mackman N. The many faces of tissue factor. J Thromb Haemost. 2009;7(Suppl 1):136–139. doi: 10.1111/j.1538-7836.2009.03368.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Day SM, Reeve JL, Pedersen B, Farris DM, Myers DD, Im M, Wakefield TW, Mackman N, Fay WP. Macrovascular thrombosis is driven by tissue factor derived primarily from the blood vessel wall. Blood. 2005;105:192–198. doi: 10.1182/blood-2004-06-2225. [DOI] [PubMed] [Google Scholar]
  • 3.Niessen F, Schaffner F, Furlan-Freguia C, Pawlinski R, Bhattacharjee G, Chun J, Derian CK, Andrade-Gordon P, Rosen H, Ruf W. Dendritic cell PAR1-S1P3 signalling couples coagulation and inflammation. Nature. 2008;452:654–658. doi: 10.1038/nature06663. [DOI] [PubMed] [Google Scholar]
  • 4.Hembrough TA, Swartz GM, Papathanassiu A, Vlasuk GP, Rote WE, Green SJ, Pribluda VS. Tissue factor/factor VIIa inhibitors block angiogenesis and tumor growth through a nonhemostatic mechanism. Cancer Res. 2003;63:2997–3000. [PubMed] [Google Scholar]
  • 5.Smith NL, Huffman JE, Strachan DP, Huang J, Dehghan A, Trompet S, Lopez LM, Shin SY, Baumert J, Vitart V, Bis JC, Wild SH, Rumley A, Yang Q, Uitterlinden AG, Stott DJ, Davies G, Carter AM, Thorand B, Polasek O, McKnight B, Campbell H, Rudnicka AR, Chen MH, Buckley BM, Harris SE, Peters A, Pulanic D, Lumley T, de Craen AJ, Liewald DC, Gieger C, Campbell S, Ford I, Gow AJ, Luciano M, Porteous DJ, Guo X, Sattar N, Tenesa A, Cushman M, Slagboom PE, Visscher PM, Spector TD, Illig T, Rudan I, Bovill EG, Wright AF, McArdle WL, Tofler G, Hofman A, Westendorp RG, Starr JM, Grant PJ, Karakas M, Hastie ND, Psaty BM, Wilson JF, Lowe GD, O’Donnell CJ, Witteman JC, Jukema JW, Deary IJ, Soranzo N, Koenig W, Hayward C. Genetic predictors of fibrin D-dimer levels in healthy adults. Circulation. 2011;123:1864–1872. doi: 10.1161/CIRCULATIONAHA.110.009480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Smith NL, Hindorff LA, Heckbert SR, Lemaitre RN, Marciante KD, Rice K, Lumley T, Bis JC, Wiggins KL, Rosendaal FR, Psaty BM. Association of genetic variations with nonfatal venous thrombosis in postmenopausal women. JAMA. 2007;297:489–498. doi: 10.1001/jama.297.5.489. [DOI] [PubMed] [Google Scholar]
  • 7.Cushman M, Tsai AW, White RH, Heckbert SR, Rosamond WD, Enright P, Folsom AR. Deep vein thrombosis and pulmonary embolism in two cohorts: the longitudinal investigation of thromboembolism etiology. Am J Med. 2004;117:19–25. doi: 10.1016/j.amjmed.2004.01.018. [DOI] [PubMed] [Google Scholar]
  • 8.Germain M, Saut N, Greliche N, Dina C, Lambert JC, Perret C, Cohen W, Oudot-Mellakh T, Antoni G, Alessi MC, Zelenika D, Cambien F, Tiret L, Bertrand M, Dupuy AM, Letenneur L, Lathrop M, Emmerich J, Amouyel P, Tregouet DA, Morange PE. Genetics of venous thrombosis: insights from a new genome wide association study. PLoS One. 2011;6:e25581. doi: 10.1371/journal.pone.0025581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Tregouet DA, Heath S, Saut N, Biron-Andreani C, Schved JF, Pernod G, Galan P, Drouet L, Zelenika D, Juhan-Vague I, Alessi MC, Tiret L, Lathrop M, Emmerich J, Morange PE. Common susceptibility alleles are unlikely to contribute as strongly as the FV and ABO loci to VTE risk: results from a GWAS approach. Blood. 2009;113:5298–5303. doi: 10.1182/blood-2008-11-190389. [DOI] [PubMed] [Google Scholar]
  • 10.Heit JA, Matsumoto ME, Asmann Y, Jeavons E, Armasu SM, Petterson TM, Cunningham JM, Andrade M. Imputed Genome Wide and Candidate Gene Studies Reveals Novel Genetic Variant Associated with Venous Thromboembolism (VTE) Genet Epidemiol. 2010;34:974. [Google Scholar]
  • 11.Hofman A, Breteler MM, van Duijn CM, Krestin GP, Pols HA, Stricker BH, Tiemeier H, Uitterlinden AG, Vingerling JR, Witteman JC. The Rotterdam Study: objectives and design update. Eur J Epidemiol. 2007;22:819–829. doi: 10.1007/s10654-007-9199-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Ridker PM, Chasman DI, Zee RY, Parker A, Rose L, Cook NR, Buring JE. Rationale, design, and methodology of the Women’s Genome Health Study: a genome-wide association study of more than 25,000 initially healthy american women. Clin Chem. 2008;54:249–255. doi: 10.1373/clinchem.2007.099366. [DOI] [PubMed] [Google Scholar]
  • 13.Fried LP, Borhani NO, Enright P, Furberg CD, Gardin JM, Kronmal RA, Kuller LH, Manolio TA, Mittelmark MB, Newman A, O’Leary DH, Psaty BM, Rautaharju P, Tracy RP, Weiler PG. The Cardiovascular Health Study: design and rationale. Ann Epidemiol. 1991;1:263–276. doi: 10.1016/1047-2797(91)90005-w. [DOI] [PubMed] [Google Scholar]
  • 14.The Atherosclerosis Risk in Communities (ARIC) Study: design and objectives. The ARIC investigators. Am J Epidemiol. 1989;129:687–702. [PubMed] [Google Scholar]
  • 15.Cushman M, Folsom AR, Wang L, Aleksic N, Rosamond WD, Tracy RP, Heckbert SR. Fibrin fragment D-dimer and the risk of future venous thrombosis. Blood. 2003;101:1243–1248. doi: 10.1182/blood-2002-05-1416. [DOI] [PubMed] [Google Scholar]
  • 16.Ye Z, Liu EH, Higgins JP, Keavney BD, Lowe GD, Collins R, Danesh J. Seven haemostatic gene polymorphisms in coronary disease: meta-analysis of 66,155 cases and 91,307 controls. Lancet. 2006;367:651–658. doi: 10.1016/S0140-6736(06)68263-9. [DOI] [PubMed] [Google Scholar]
  • 17.Lange LA, Reiner AP, Carty CL, Jenny NS, Cushman M, Lange EM. Common genetic variants associated with plasma fibrin D-dimer concentration in older European- and African-American adults. J Thromb Haemost. 2008;6:654–659. doi: 10.1111/j.1538-7836.2008.02906.x. [DOI] [PubMed] [Google Scholar]
  • 18.Bertina RM, Koeleman BP, Koster T, Rosendaal FR, Dirven RJ, de Ronde H, van der Velden PA, Reitsma PH. Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature. 1994;369:64–67. doi: 10.1038/369064a0. [DOI] [PubMed] [Google Scholar]
  • 19.Dahlback B, Hildebrand B. Inherited resistance to activated protein C is corrected by anticoagulant cofactor activity found to be a property of factor V. Proc Natl Acad Sci U S A. 1994;91:1396–1400. doi: 10.1073/pnas.91.4.1396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Carter AM, Catto AJ, Kohler HP, Ariens RA, Stickland MH, Grant PJ. alpha-fibrinogen Thr312Ala polymorphism and venous thromboembolism. Blood. 2000;96:1177–1179. [PubMed] [Google Scholar]
  • 21.Reiner AP, Carty CL, Carlson CS, Wan JY, Rieder MJ, Smith JD, Rice K, Fornage M, Jaquish CE, Williams OD, Tracy RP, Lewis CE, Siscovick DS, Boerwinkle E, Nickerson DA. Association between patterns of nucleotide variation across the three fibrinogen genes and plasma fibrinogen levels: the Coronary Artery Risk Development in Young Adults (CARDIA) study. J Thromb Haemost. 2006;4:1279–1287. doi: 10.1111/j.1538-7836.2006.01907.x. [DOI] [PubMed] [Google Scholar]
  • 22.Standeven KF, Grant PJ, Carter AM, Scheiner T, Weisel JW, Ariens RA. Functional analysis of the fibrinogen Aalpha Thr312Ala polymorphism: effects on fibrin structure and function. Circulation. 2003;107:2326–2330. doi: 10.1161/01.CIR.0000066690.89407.CE. [DOI] [PubMed] [Google Scholar]

RESOURCES