title: Characterization of F8 defects in haemophilia A in Pakistan: investigation of correlation between mutation type and the in vitro thrombin generation assay.
abstract: Hereditary haemophilia A is an X-linked bleeding disorder caused by mutations in the coagulation factor VIII gene (FVIII abbreviates protein, gene symbol F8). The mutation spectrum has been reported in various populations but not in Pakistan. The aims of this study were to (i) characterize F8 mutations in a large haemophilia A cohort from Pakistan and to (ii) investigate whether in vitro thrombin generation (TG) differs according to mutation type (null compared with missense) in severe haemophilia A. One hundred individuals diagnosed with haemophilia A and 100 healthy controls were recruited in Pakistan. Phenotypic measurements were re-evaluated in Cardiff; the essential regions of F8 were screened for the causative defect. A diagnosis of haemophilia A was confirmed for 92 individuals, 7 were found to have haemophilia B and 1 did not have haemophilia. The F8 defects were characterized for 80 of the 92 haemophilia A individuals and comprised point mutations, inversions (intron 22 and intron 1) and frameshifts. Point mutations (41%) were the most frequent, followed by the intron 22 inversion (20%). Thirty novel variants were identified. Comparison of in vitro TG parameters [velocity index (VI) and peak] was made between severe individuals who had a null mutation (no FVIII) and those with a missense change (dysfunctional FVIII), no significant difference was observed. The spectrum of F8 defects in Pakistan is heterogenous; VI and peak in severe haemophilia A are not influenced by whether the underlying mutation gives rise to dysfunctional FVIII or no coagulation factor at all.
authors.affiliation: Department of Haematology, School of Medicine, Cardiff University, Cardiff, UK.
authors.forename: F
authors.initials: F
authors.lastname: Khanum
authors.forename: P W
authors.initials: PW
authors.lastname: Collins
authors.forename: R L
authors.initials: RL
authors.lastname: Harris
authors.forename: D J
authors.initials: DJ
authors.lastname: Bowen
mesh_terms: D001780:Blood Coagulation Tests; D005091:Exons; D005169:Factor VIII / Q000235:genetics*; D006239:Haplotypes; D006467:Hemophilia A / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007438:Introns; D008297:Male; D009154:Mutation; D010154:Pakistan; D010641:Phenotype; D012720:Severity of Illness Index; D013917:Thrombin / Q000378:metabolism*
chemical_list: D005169:Factor VIII; D013917:Thrombin
keywords: F8; Pakistan; haemophilia A; intron 22 inversion; mutation; thrombin generation