Molecular methods in thrombophilic states diagnostics

Česká verzia

Authors: L. Slavík; V. Krčová; A. Hluší; J. Procházková; J. Úlehlová
Authors place of work: Hemato-onkologická klinika Lékařské fakulty UP a FN Olomouc, přednosta prof. MUDr. Karel Indrák, DrSc.
Published in the journal: Vnitř Lék 2009; 55(3): 302-309
Category: 15th Parizek's Days

Summary

Molecular genetic methods passed into the field of investigation of thrombophilic states in 90th years of last century, along with the first discoveries of coagulation inhibitors (AT III, protein C and protein S). They have acquired a widespread use above all with the detection of the molecular basis of activated protein C (APC) resistance in 1994 by prof. Bertina. At the present time, a wide range of molecular genetic markers, linked with a clearly documented increased risk of thrombophilia are adapted. They include mutations of factor V Leiden 506R/Q, of protrombin 20210G/A, MTHFR 677C/T in homozygous form, mutation of PAI‑1 4G/5G, mutations of different coagulation inhibitors and finally a range of polymorphisms with still not precisely defined increased risk for thrombophilia (F XIII Val34leu, platelets glycopeproteins, endothelial protein C receptor and trombomodulin). From the methodological viewpoint, all these techniques are based on the principle polymerase chain reaction (PCR). In the last period of time, however there was a rapid evolution, allowing a significant improvement in their laboriousness. Nowadays, splitting with the aid of restriction endonucleases, real time PCR or allel specific primers for PCR. The second, where molecular genetic methods are currently under use, is pathophysiological investigation of the single coagulation processes. Here, in a fact, most significant progress has been in the field of APC resistance made elucidation. Although still in the 90th years of the past century the genetical cause of these coagulation disturbance was unequivocally documented its clinically heterozygous appears not yet fully understood at the moment. Similarly, in prothrombin mutation, only the latest investigations have outlined the probable mechanism of expression. Concerning the future evolution of molecular genetic methods, there can be observed a clear cut tendency to better understanding the pathophysiologic cause of thrombophilia in comparison with the searching for new coagulation defects which consecutively bear lesser a relative risk of thrombosis.

Key words:
thrombophilia – genetic markers of thrombophilia – F V Leiden – protrombin mutation

Zdroje

1. Rees DC, Cox M, Clegg JB. World distribution of factor V Leiden. Lancet 1995; 346: 1133–1134.

2. Rosendaal FR, Koster T, Vandenbroucke JP et al. High risk of thrombosis in patients homozygous for factor V Leiden (activated protein C resistance). Blood 1995; 85: 1504–1508.

3. Ridker PM, Miletich JP, Hennekens CH et al. Ethnic distribution of factor V Leiden in 4047 men and women. JAMA 1997; 277: 1305–1307.

4. Vorlova Z, Hrachovinova I, Matyskova M. Probability of thrombosis in patients with factor V Leiden. Thromb Haemost 1997; 78: 309.

5. Chrobák L, Dulíček P. Resistance to activated protein C as pathogenic factor of venous thromboembolism. Hradec Králové: Acta Medica 1996; 39: 55–62.

6. Dulíček P, Šafářová M, Chrobák L. Mutace FV Leiden – nejčastější rizikový faktor pro vznik žilní trombózy. Hematológia a transfuziológia 1997; 4: 6–9.

7. Bertina RM, Koeleman RPC, Koster T et al. Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature 1994; 369: 64–67.

8. Dahlbäck B, Carlsson M, Svensson PJ. Familial thrombophilia due to a previously unrecognised mechanism characterized by poor anticoagulant response to activated protein C: prediction of a cofactor to activated protein C. Proc Natl Acad Sci USA 1993; 90: 1004–1008.

9. Koster T, Rosendaal FR, De Ronde H et al. Venous thrombosis due to a poor anticoagulant response to activated protein C: Leiden Thrombophilia Study. Lancet 1993; 342: 1503–1506.

10. Ridker PM, Hennekens CH, Lindpainter K et al. Mutation in the gene coding for coagulation factor V and the risk of myocardial infarction, stroke, and venous thrombosis in apparently healthy men. N Engl J Med 1995; 332: 912–917.

11. Anderson FA, Wheeler HB, Goldberg RJ et al. A population based perspective of the hospital incidence and case-fatality rates of deep vein thrombosis and pulmonary embolism. The Worcester DVT study. Arch Intern Med 1991; 151: 933–938.

12. Poort SR, Rosendaal FR, Reitsma PH et al. A commongenetic variation in the 3’-untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increase in venous thrombosis. Blood 1996; 88: 3698–3703.

13. Rosendaal FR, Doggen CJM, Zivelin A et al. Geographic distribution of the 20210 G to A prothrombin variant. Thromb Haemost 1998; 79: 706–708.

14. Matýšková M, Buliková A, Šlechtová M et al. The prevalence of the prothrombin mutation 20210A in Brno. XV meeting of the ISH – African and European division. Final programme and abstracts, 124.

15. Souto JC, Coll I, Llobet D et al. The prothrombin 20210A allele is the most present genetic risk factor for venous thromboembolism in the Spanish population. Thromb Haemost 1998; 80: 366–369.

16. Falcon CR, Cattaneo M, Panzeri D et al. High prevalence of hyperhomocyst(e)inemia in patients with juvenile venous thrombosis. Arterioscler Thromb 1994; 14: 1080–1083.

17. Den Heijer M, Koster T, Blom HJ et al. Hyperhomocysteinemia as a risk factor for deep-vein thrombosis. N Engl J Med 1996; 334: 759–762.

18. Simioni P, Prandoni P, Burlina A et al. Hyperhomocysteinemia and deep-vein thrombosis: a case-control study. Thromb Haemost 1996; 76: 883–886.

19. Hyánek J, Hoffman R. Hyperhomocysteinémie a její diagnostický význam u cévních onemocnění. Praktická flebologie 1997; 2: 61–71.

20. D’Angelo A, Selhub J. Homocysteine and thrombotic disease. Blood 1997; 90: 1–11.

21. Kang SS, Wong PWK, Norusis M. Homocysteinemia due to folate deficiency. Metabolism 1987; 36: 458–462.

22. Kang SS, Wong PWK, Norusis M. Homocysteinemia due to folate deficiency. Metabolism 1987; 36: 458–462.

23. Rees MM, Rodgers GM. Homocysteinemia: association of a metabolic disorder with vascular disease and thrombosis. Thrombosis Research 1993; 71: 337–359.

24. Ubbink JB, Vermaak WJ, Van der Merwe A et al. Vitamin B12, vitamin B6, and folate nutritional status in men with hyperhomocysteinemia. Am J Clin Nutr 1993; 57: 47–53.

25. Mudd SH, Skovby F, Levy HL et al. The natural history of homocystinuria due to cystathionine beta‑synthase deficiency. Am J Hum Genet 1985; 37: 1–31.

26. Engbertsen AMT, Franken DG, Boers GHJ et al. Thermolabile 5,10-methylenetetrahydrofolate reductase as a cause of mild hyperhomocysteinemia. Am J Hum Genet 1995; 56: 142–150.

27. Frosst P, Blom HJ, Milos R et al. A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet 1995; 10: 111–113.

28. Egeberg O. Inherited antithrombin deficiency causing thrombophilia. Thromb Diath Haemorrh 1965; 13: 516–530.

29. Thaler E, Lechner K. Antithrombin III deficiency and thromboembolism. Clin Haematol 1981; 10: 369–390.

30. Demers C, Ginsberg JS, Hirsh J et al. Thrombosis in antithrombin III deficient persons: report of a large kindred and literature review. Ann Intern Med 1992; 116: 754–761.

31. Hirsh J, Piovella F, Pini M. Congenital antithrombin III deficiency: incidence and clinical features. Am J Med 1989; 87: 34–38.

32. Lane DA, Mannucci PM, Bauer KA et al. Inherited Thrombophilia: Part 1. Thromb Haemost 1996; 76: 651–662.

33. Tait RC, Walker ID, Perry DJ et al. Prevalence of antithrombin deficiency in the healthy population. Br J Haematol 1994; 87: 106–112.

34. Bock SC et al. Assignement of the human antithrombin III structural gene to chromosome 1q23–25. Cytogenet Cell Genet 1985; 39: 67–69.

35. Lane DA et al. Antithrombin III: A database of mutations. Thrombos Haemostasis 1991; 66: 657–661.

36. Griffin JH, Evatt B, Zimmerman TS et al. Deficiency of protein C in congenital thrombotic disease. J Clin Invest 1981; 68: 1370–1373.

37. Allaart CF, Poort SR, Rosendaal FR et al. Increased risk of venous thrombosis in carriers of protein C deficiency defect. Lancet 1993; 341: 134–138.

38. Broekmans AW, Veltkamp JJ, Bertina RM. Congenital protein C deficiency and venous thromboembolism: a study of three Dutchfamilies. N Engl J Med 1983; 309: 340–344.

39. Allaart CF, Poort SR, Rosendaal FR et al. Increased risk of venous thrombosis in carriers of protein C deficiency defect. Lancet 1993; 341: 134–138.

40. Koeleman BPC, Reitsma PH, Allaart CF et al. APC‑resistance as an additional risk factor for thrombosis in protein C deficient families. Blood 1994; 84: 1031–1035.

41. Heijboer H, Brandjes DPM, Büller HR et al. Deficiencies of coagulation‑inhibiting and fibrinolytic proteins in outpatients with deep-vein thrombosis. N Engl J Med 1990; 323: 1512–1516.

42. Mateo J, Oliver A, Borrell M et al. The EMET Group. Laboratory evaluation and clinical characteristics of 2,132 consecutive unselected patients with venous thromboembolism-results of the Spanish multicentric study on thrombophilia (EMET-study). Thromb Haemost 1997; 77: 444–451.

43. Koster T, Rosendaal FR, Briët E et al. Protein C deficiency in a controlled series of unselected outpatients: an infrequent but clear risk factor for venous thrombosis (Leiden Thrombophilia Study). Blood 1995; 85: 2756–2761.

44. Reitsma PH, Bernardi F, Doig RG et al. Protein C deficiency a database of mutations. 1995 Update. Thromb Haemost 1995; 73: 876–889.

45. Walker ID et al. Guidelines on the Investigation and Management of Thrombophilia. J Clin Path 1990; 43: 703–709.

46. Cripe LD, Moore KD, Kane WH. Structure of the gene for human coagulation factor V. Biochemistry 1992; 31: 3777–3785.

47. Jenny RJ, Pittman DD, Toole JJ et al. Complete cDNA and derived amino acid sequence of human factor V. Proc Natl Acad Sci USA 1987; 84: 4846–4850.

48. Tracy PB, Eide LL, Bowie EJ et al. Radioimmunoassay of factor V in human plasma and platelets. Blood 1982; 60: 59–63.

49. Suehiro Y, Veljkovic DK, Fuller N et al. Endocytosis and storage of plasma factor V by human megakaryocytes. Thromb Haemost 2005; 94: 585–592.

50. Hoekema L, Nicolaes GAF, Hemker HC et al. Human factor Va1 and factor Va2: properties in the procoagulant and anticoagulant pathways. Biochemistry 1997; 36: 3331–3335.

51. Esmon CT. The subunit structure of thrombin‑activated factor V. Isolation of activated factor V, separation of subunits, and reconstitution of biological activity. J Biol Chem 1979; 254: 964–973.

52. Husten EJ, Esmon CT, Johnson AE. The active site of blood coagulation factor Xa. Its distance from the phospholipid surface and its conformational sensitivity to components of the prothrombinase complex. J Biol Chem 1987; 262: 12953–12961.

53. Yegneswaran S, Fernandez JA, Griffin JH et al. Factor Va increases the affinity of factor Xa for prothrombin: a binding study using a novel photoactivable thiol-specific fluorescent probe. Chem Biol 2002; 9: 485–494.

54. Dahlbäck B, Carlsson M, Svensson PJ. Familial thrombophilia due to a previously unrecognized mechanism characterized by poor anticoagulant response to activated protein C: prediction of a cofactor to activated protein C. Proc Natl Acad Sci USA 1993; 90: 1004–1008.

55. Bertina RM, Koeleman BP, Koster T et al. Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature 1994; 369: 64–67.

56. Leroy-Matheron C, Mallat A, Duvoux C et al. Inhibitor against coagulation factor V after liver transplantation. Transplantation 1999; 68: 1054–1056.

57. Gehring NH, Frede U, Neu-Yilik G et al. Increased efficiency of mRNA 3 end formation: a new genetic mechanism contributing to hereditary thrombophilia. Nat Genet 2001; 28: 389–392.

58. Ceelie H, Spaargaren-van Riel CC, Bertina RM et al. G20210A is a functional mutation in the prothrombin gene; effect on protein levels and 3-end formation. J Thromb Haemost 2004; 2: 119–127.

59. Danckwardt S, Gehring N, Neu-Yilik G et al. The prothrombin 3 end formation signal reveals a unique architecture that is sensitive to thrombophilic gain‑of-function mutations. Blood 2004; 104: 428–435.