title: Thrombin-activable factor X re-establishes an intrinsic amplification in tenase-deficient plasmas.
abstract: Classical hemophilia results from a defect of the intrinsic tenase complex, the main factor X (FX) activator. Binding of factor VIIa to tissue factor triggers coagulation, but little amplification of thrombin production occurs. Handling of hemophilia by injection of the deficient or missing (thus foreign) factor often causes immunological complications. Several strategies have been designed to bypass intrinsic tenase complex, but none induce true auto-amplification of thrombin production. In an attempt to re-establish a cyclic amplification of prothrombin activation in the absence of tenase, we prepared a chimera of FX having fibrinopeptide A for the activation domain (FX(FpA)). We reasoned that cascade initiation would produce traces of thrombin that would activate FX(FpA) (contrary to its normal homologue). Given that the activation domain of FX is released upon activation, thrombin cleavage would produce authentic FXa that would produce more thrombin, which in turn would activate more chimeras. FX(FpA) was indeed activable by thrombin, albeit at a relatively low rate (5 x 10(3) M(-1) s(-1)). Nevertheless, FX(FpA) allowed in vitro amplification of thrombin production, and 100 nM efficiently corrected thrombin generation in tenase-deficient plasmas. A decisive advantage of FX(FpA) could be that the artificial cascade is self-regulating: FX(FpA) had little influence on the clotting time of normal plasma, yet corrected that of tenase deficiency. Another advantage could be the half-life of FX(FpA) in blood; FX has a half-life of about 30 h (less than 3 h for FVIIa). It is also reasonable to expect little or no immunogenicity, because FX and fibrinopeptide A both circulate normally in the blood of hemophiliacs.
authors.affiliation: INSERM U428, Faculté de Pharmacie, Université Paris V, 75270 Paris, France.
authors.forename: Virginie B
authors.initials: VB
authors.lastname: Louvain-Quintard
authors.forename: Elsa P
authors.initials: EP
authors.lastname: Bianchini
authors.forename: Claire
authors.initials: C
authors.lastname: Calmel-Tareau
authors.forename: Madjid
authors.initials: M
authors.lastname: Tagzirt
authors.forename: Bernard F
authors.initials: BF
authors.lastname: Le Bonniec
mesh_terms: D001777:Blood Coagulation; D003029:Coagulants / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000378:metabolism / Q000502:physiology*; D018076:DNA, Complementary / Q000378:metabolism; D004220:Disulfides; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D005170:Factor X / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology*; D005344:Fibrinopeptide A / Q000737:chemistry / Q000378:metabolism; D006467:Hemophilia A / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D008956:Models, Chemical; D009363:Neoplasm Proteins / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013917:Thrombin / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors
chemical_list: D003029:Coagulants; D018076:DNA, Complementary; D004220:Disulfides; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D005344:Fibrinopeptide A; D005170:Factor X; D013917:Thrombin; D003546:Cysteine Endopeptidases; C030910:cancer procoagulant