The factor V Glu1608Lys mutation is recurrent in familial thrombophilia

• Published in: Journal of thrombosis and haemostasis 2005-09, Vol.3 (9), p.2032-2038
• Main Authors: LUNGHI, B., SCANAVINI, D., CASTOLDI, E., GEMMATI, D., TOGNAZZO, S., REDAELLI, R., GHIRARDUZZI, A., IERAN, M., PINOTTI, M., BERNARDI, F.
• Format: Article
• Language: English
• Subjects: Case-Control Studies; DNA Mutational Analysis; Factor V - genetics; factor V deficiency; Factor V Deficiency - genetics; factor V mutation; Family Health; Female; Haplotypes; Heterozygote; Humans; Incidence; Leukocytes - chemistry; Male; Mutation, Missense; Point Mutation; pseudo‐homozygosity; Receptors, Cell Surface - genetics; RNA, Messenger - analysis; thrombophilia; Thrombophilia - genetics
• ISSN: 1538-7933; 1538-7836; 1538-7836
• DOI: 10.1111/j.1538-7836.2005.01453.x

Background: Co‐inheritance of heterozygous factor V deficiency with FV Leiden enhances the activated protein C resistance (APCR) associated with this mutation, resulting in pseudo‐homozygous APCR. The role of FV deficiency in modulating thrombotic risk in this rare condition is poorly understood. Methods and results: We have identified in thrombophilic patients with FV deficiency a novel FV gene mutation (c. 4996G>A), predicting the Glu1608Lys substitution in the A3 domain. The heterozygous mutation was detected in three unrelated patients, two carriers of the FV Leiden mutation, and one of the FVHR2 haplotype. The Glu1608Lys change was also present in two subjects with mild FV deficiency, and absent in 200 controls. The FV1608Lys carriers showed reduced mean FV activity (42% ± 12%) and antigen (53% ± 18%) levels and, in Western blot analysis, reduced amounts of intact platelet FV. The restriction fragment length polymorphism (RFLP) study identified two haplotypes underlying the mutation, which suggests that it is recurrent. In heterozygous subjects the amount of FV1608Lys mRNA in white blood cells was similar to that produced by the counterpart alleles (FVWt or FVHR2). Recombinant FV1608Lys (rFV1608Lys), detected by Western blot in the conditioned medium, was indistinguishable from rFVWt and FV antigen and activity were found to be respectively 44% ± 20% and 13% ± 4% of rFVWt. Conclusions: Our data indicate that FVGlu1608Lys predicts a CRM (plasma)/CRMred (cell culture) FV deficiency, and may contribute to thrombophilia in carriers of FV Leiden and FVHR2 haplotype via a pseudo‐homozygosity mechanism. Our findings help to define the molecular bases of FV deficiency and thrombophilia.