Molecular Characterization of a Type I Quantitative Factor V Deficiency in a Thrombosis Patient that Is “Pseudo Homozygous” for Activated Protein C Resistance

Joan F Guasch; Ruud P M Lensen; Rogier M Bertina

doi:10.1055/s-0038-1655948

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1997; 77(02): 252-257
DOI: 10.1055/s-0038-1655948

Original Article

Schattauer GmbH Stuttgart

Molecular Characterization of a Type I Quantitative Factor V Deficiency in a Thrombosis Patient that Is “Pseudo Homozygous” for Activated Protein C Resistance

Joan F Guasch

The Haemostasis and Thrombosis Research Center, University Hospital Leiden, The Netherlands

,

Ruud P M Lensen

¹Department of Clinical Epidemiology, University Hospital Leiden, The Netherlands

,

Rogier M Bertina

The Haemostasis and Thrombosis Research Center, University Hospital Leiden, The Netherlands

› Author Affiliations

Abstract

PDF Download

Summary

Resistance to activated protein C (APC), which is associated with the FV Leiden mutation in the large majority of the cases, is the most common genetic risk factor for thrombosis. Several laboratory tests have been developed to detect the APC-resistance phenotype. The result of the APC-resistance test (APC-sensitivity ratio, APC-SR) usually correlates well with the FV Leiden genotype, but recently some discrepancies have been reported. Some thrombosis patients that are heterozygous for FV Leiden show an APC-SR usually found only in homozygotes for the defect. Some of those patients proved to be compound heterozygotes for the FV Leiden mutation and for a type I quantitative factor V deficiency. We have investigated a thrombosis patient characterized by an APC-SR that would predict homozygosity for FV Leiden. DNA analysis showed that he was heterozygous for the mutation. Sequencing analysis of genomic DNA revealed that the patient also is heterozygous for a G⁵⁵⁰⁹→A substitution in exon 16 of the factor V gene. This mutation interferes with the correct splicing of intron 16 and leads to the presence of a null allele, which corresponds to the “non-FV Leiden” allele. The conjunction of these two defects in the patient apparently leads to the same phenotype as observed in homozygotes for the FV Leiden mutation.

PDF (435 kb)

References

References
1 Koster T, Rosendaal FR, de Ronde H, Briët E, Vandenbroucke JP, Bertina RM. Venous thrombosis due to poor anticoagulant response to activated protein C: Leiden Thrombophilia Study. Lancet 1993; 342: 1503-1506
2 Griffin JH, Evatt B, Wideman C, Fernandez JA. Anticoagulant protein C pathway defective in majority of thrombophilic patients. Blood 1993; 82: 1989-1993
3 Halbmayer WM, Haushofer A, Schon R, Fischer M. The prevalence of poor anticoagulant response to activated protein C (APC resistance) among patients suffering from stroke or venous thrombosis and among healthy subjects. Blood Coag Fibrinol 1994; 5: 51-57
4 Svensson PJ, Dahlbäck B. Resistance to activated protein C as a basis for venous thrombosis. N Engl J Med 1994; 330: 517-522
5 Legnani C, Palareti G, Biagi R, Coccheri S. Activated protein C resistance in deep-vein thrombosis. Lancet 1994; 343: 541-542
6 Cadroy Y, Sié P, Boneu B. Frequency of a defective response to activated protein C in patients with a history of venous thrombosis. Blood 1994; 83: 2008-2009
7 Cushman M, Bhushan F, Bovill E, Tracy R. Plasma resistance to activated protein C in venous and arterial thrombosis. Thromb Haemost 1994; 72: 647-657
8 Dahlbäck B. Physiological anticoagulation. Resistance to activated protein C and venous thromboembolism. J Clin Invest 1994; 94: 923-927
9 Bertina RM, Koeleman BPC, Koster T, Rosendaal FR, Dirven RJ, deRonde H, van der VeldenPA, Reitsma PH. Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature 1994; 369: 64-67
10 Greengard JS, Sun X, Xu X, Fernandez JA, Griffin JH, Evatt B. Activated protein C resistance caused by Arg506Gln mutation in factor Va. Lancet 1994; 343: 1361-1362
11 Voorberg J, Roelse J, Koopman R, Büller H, Berends F, ten Cate JW, Mertens K, van MourikJA. Association of idiopathic venous thromboembolism with single point mutation at ARG506 of factor V. Lancet 1994; 343: 1535-1536
12 Zӧller B, Dahlbäck B. Linkage between inherited resistance to activated protein C and factor V gene mutation in venous thrombosis. Lancet 1994; 343: 1536-1538
13 Zöller B, Svensson PJ, He X, Dahlbäck B. Identification of the same factor V gene mutation in 47 out of 50 thrombosis-prone families with inherited resistance to activated protein C. J Clin Invest 1994; 94: 2521-2524
14 Kalafatis M, Bertina RM, Rand MD, Mann KG. Characterization of the molecular defect in factor VR506Q. J Biol Chem 1995; 270: 4053-4057
15 Nicolaes GA, Tans G, Thomassen MC, Hemker HC, Pabinger I, Varadi K, Schwarz HP, Rosing J. Peptide bond cleavages and loss of functional activity during inactivation of factor Va and factor VaR506Q by activated protein. J Biol Chem 1995; 270: 21158-21166
16 Rosendaal FR, Koster T, Vandenbroucke JP, Reitsma PH. High risk of thrombosis in patients homozygous for factor V Leiden (activated protein C resistance). Blood 1995; 85: 1504-1508
17 Greengard JS, Eichinger S, Griffin JH, Bauer KA. Brief report: variability of thrombosis among homozygous siblings with resistance to activated protein C due to an Arg→Gln mutation in the gene for factor V. N Engl J Med 1994; 331: 1559-1562
18 Hopmeier P, Krugluger W. Factory V Leiden and thrombophilia. N Engl J Med 1995; 332: 1381-1382
19 Simioni P, Scudeller A, Girolami A. Factor V Leiden and thrombophilia. N Engl J Med 1995; 332: 1382-1392
20 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
21 deRonde H, Bertina RM. Laboratory diagnosis of APC-resistance: a critical evaluation of the test and the development of diagnostic criteria. Thromb Haemost 1994; 72: 880-886
22 Vasse M, Leduc O, Borg JY, Chretien MH, Monconduit M. Resistance to activated protein C, evaluation of three functional assays. Thromb Res 1994; 76: 47-59
23 Sidelmann J, Gram J, Pedersen OD, Jespersen J. Influence of plasma platelets on activated protein C resistance assay. Thromb Haemost 1995; 74: 993-994
24 Greengard JS, Alhenc-Gelas M, Gandrille S, Emmerich J, Aiach M, Griffin JH. Pseudo-homozygous activated protein C resistance due to coinheritance of heterozygous factor V-R506Q and type I factor V deficiency associated with thrombosis. Thromb Haemost 1995; 73: 1361
25 Simioni P, Scudeller A, Radossi P, Gavasso S, Girolami B, Tormene D, Girolami A. “Pseudo Homozygous” activated protein C resistance due to double heterozygous factor V defects (factor V Leiden mutation and type I quantitative factor V defect) associated with thrombosis: report of two cases belonging to two unrelated kindreds. Thromb Haemost 1996; 75: 422-426
26 Zehnder JL, Jain M. Recurrent thrombosis due to compound heterozygosity for factor V Leiden and factor V deficiency. Blood Coag Fibrinol 1996; 07: 361-362
27 Berg LP, Wieland K, Millar DS, Schlosser M, Wagner M, Kakkar VV, Reiss J, Cooper DN. Detection of a novel point mutation causing haemophilia A by PCR/direct sequencing of ectopically-transcribed factor VIII mRNA. Hum Genet 1990; 85: 655-658
28 Bertina RM, Reitsma PH, Rosendaal FR, Vandenbroucke JP. Resistance to activated protein C and factor V leiden as risk factors for venous thrombosis. Thromb Haemost 1995; 74: 449-453
29 Sun X, Evatt B, Griffin JH. Blood coagulation factor Va abnormality associated with resistance to activated protein C in venous thrombophilia. Blood 1994; 83: 3120-3125
30 Alhenc-Gelas M, Gandrille S, Aubry ML, Emmerich J, Flessinger JN, Aiach M. Unexplained thrombosis and factor V Leiden mutation. Lancet 1994; 344: 555-556
31 Beauchamp NJ, Daly ME, Hampton KK, Cooper PC, Preston FE, Peake IR. High prevalence of a mutation in the factor V gene within the U.K population: relationship to activated protein C resistance and familial thrombosis. BrJ Haematol 1994; 88: 219-222
32 Murray JM, Rand MD, Egan JO, Murphy S, Kim HC, Mann KG. Factor V New Brunswick: Ala221-to-Val substitution results in reduced cofactor <activity. Blood 1995; 86: 1820-1827
33 Kane WH, Ichinose A, Hagen FS, Davie EW. Cloning of cDNAs coding for the heavy chain region and connecting region of human factor V, a blood coagulation factor with four types of internal repeats. Biochemistry 1987; 26: 6508-6514
34 Cox MJ, Rees DC, Martinson JJ, Cleeg JB. Evidence for a single origin of factor V Leiden. Br J Haematol 1996; 92: 1022-1025
35 Jenny RJ, Pittman DD, Toole JJ, Kriz RW, Aldape RA, Hewick RM, Kaufman RJ, Mann KG. Complete cDNA and derived amino acid sequence of human factor V. Proc Natl Acad Sci USA 1987; 84: 4846-4850
36 Kane WH, Davie EW. Cloning of a cDNA coding for human factor V, a blood coagulation factor homologous to factor VIII and ceruloplasmin. Proc Natl Acad Sci USA 1986; 83: 6800-6804