Effects of plasma kallikrein deficiency on haemostasis and thrombosis in mice: Murine Ortholog of the Fletcher Trait

J. Eileen Bird; Patricia L. Smith; Xinkang Wang; William A. Schumacher; Frank Barbera; Jean-Pierre Revelli; Dietmar Seiffert

doi:10.1160/TH11-10-0682

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2012; 107(06): 1141-1150
DOI: 10.1160/TH11-10-0682

Animal Models

Schattauer GmbH

Effects of plasma kallikrein deficiency on haemostasis and thrombosis in mice: Murine Ortholog of the Fletcher Trait

J. Eileen Bird

¹Department of Thrombosis Biology, Bristol-Myers Squibb, Pennington, New Jersey, USA

,

Patricia L. Smith

¹Department of Thrombosis Biology, Bristol-Myers Squibb, Pennington, New Jersey, USA

,

Xinkang Wang

²Agennix, Inc., Princeton, New Jersey, USA

,

William A. Schumacher

¹Department of Thrombosis Biology, Bristol-Myers Squibb, Pennington, New Jersey, USA

,

Frank Barbera

¹Department of Thrombosis Biology, Bristol-Myers Squibb, Pennington, New Jersey, USA

,

Jean-Pierre Revelli

³Lexicon Pharmaceuticals, The Woodlands, Texas, USA

,

Dietmar Seiffert

¹Department of Thrombosis Biology, Bristol-Myers Squibb, Pennington, New Jersey, USA

› Author Affiliations

Abstract

Summary

Plasma kallikrein is a multifunctional serine protease involved in contact activation of coagulation. Deficiency in humans is characterised by prolonged activated partial thromboplastin time (aPTT); however, the balance between thrombosis and haemostasis is not fully understood. A study of plasma kallikrein-deficient mice revealed increased aPTT, without prolonged bleeding time. Prekallikrein antisense oligonucleotide (ASO) treatment in mice suggested potential for a positive therapeutic index. The current goal was to further define the role of plasma kallikrein in coagulation. Blood pressure and heart rate were normal in plasma kallikrein-deficient mice, and mice were completely protected from occlusion (100 ± 1.3% control flow) in 3.5% FeCl₃ -induced arterial thrombosis versus heterozygotes (20 ± 11.4%) and wild-type littermates (8 ± 0%). Vessels occluded in 8/8 wild-type, 7/8 heterozygotes, and 0/8 knockouts. Anti-thrombotic protection was less pronounced in 5% FeCl₃-induced arterial injury. Integrated blood flow was 8 ± 0% control in wild-type and heterozygotes, and significantly (p<0.01) improved to 43 ± 14.2% in knockouts. The number of vessels occluded was similar in all genotypes. Thrombus weight was significantly reduced in knockouts (−47%) and heterozygotes (−23%) versus wild-type in oxidative venous thrombosis. Average tail bleeding time increased modestly in knockout mice compared to wild-type. Average renal bleeding times were similar in all genotypes. These studies confirm and extend studies with prekallikrein ASO, and demonstrate that plasma kallikrein deletion prevents occlusive thrombus formation in mice with a minimal role in provoked bleeding. Additional support for the significance of the intrinsic pathway in the coagulation cascade is provided, as well as for a potential new anti-thrombotic approach.

Keywords

Protease - animal model - gene knockout

Full Text

References

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