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Thromb Haemost 2008; 100(06): 1111-1122
DOI: 10.1160/TH08-04-0242
Platelets and Blood Cells
Schattauer GmbH

Altered bioavailability of platelet-derived factor VIII during thrombocytosis reverses phenotypic efficacy in haemophilic mice

Andrea L. Damon
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
2   Program in Genetics, State University of New York, Stony Brook, New York, USA
,
Lesley E. Scudder
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
,
Dmitri V. Gnatenko
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
,
Varsha Sitaraman
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
2   Program in Genetics, State University of New York, Stony Brook, New York, USA
,
Patrick Hearing
2   Program in Genetics, State University of New York, Stony Brook, New York, USA
3   Dept. Microbiology and Molecular Genetics, State University of New York, Stony Brook, New York, USA
,
Jolyon Jesty
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
,
Wadie F. Bahou
1   Department of Medicine, State University of New York, Stony Brook, New York, USA
2   Program in Genetics, State University of New York, Stony Brook, New York, USA
› Author Affiliations
Further Information

Publication History

Received: 18 April 2008

Accepted after major revision: 12 September 2008

Publication Date:
23 November 2017 (online)

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Summary

Ectopic delivery of factor VIII (FVIII) to megakaryocytes (Mk) represents a viable approach for localized tenase generation by concentrating the FVIIIa/FIXa enzymecofactor complex onto activated platelet membranes. We utilized a core rat platelet factor 4 (PF4) promoter for Mk/platelet-restricted expression of human B-domain-deleted (hBDD) FVIII within the background of a haemophilia A mouse (rPF4/hBDD/FVIII-/-). Platelets from rPF4/hBDD/FVIII-/- mice contained ∼122 mU FVIII:C/1x109 platelets/ml with no detectable plasmatic FVIII:C,and with no effect on α-granule-derived platelet factorV/Va function.Paired tenase assays (± thrombin) confirmed that platelet (pt) FVIII (unlike platelet FV) required thrombin cleavage for complete activation. rPF4/hBDD/FVIII-/- mice exposed to a thrombocytotic stimulus (thrombopoietin, TPO) demonstrated a statistically-significant 66% reduction in molar ptFVIII activity with a non-significant reduction in total ptFVIII biomass. Decreased molar ptFVIII concentration correlated with loss of phenotypic correction as evaluated using a haemostatic tail-snip assay. Comparative studies using a transgenic mouse expressing human amyloid-β-precursor protein (hAβPP) from the rPF4 promoter confirmed diminished hAβPP expression without affecting endogenous α-granule PF4, establishing generalizability of these observations.While Mk/plateletreleased ptFVIII (unlike pFV) is proteolytically inactive, we also conclude that thrombocytotic stimuli negatively affect ptFVIII bioavailability and phenotypic efficacy, results which correlate best with molar ptFVIII concentration, and not systemically available ptFVIII.

Keywords

Animal models - gene targeting - coagulation factors
 

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