Inhibition of intravascular thrombosis in murine endotoxemia by targeted expression of hirudin and tissue factor pathway inhibitor analogs to activated endothelium.
We have generated transgenic mice expressing the leech anticoagulant hirudin and human tissue factor pathway inhibitor tethered to the cell surface by fusion with fragments of human CD4 and P-selectin. Expression of the transgenes is under the control of the CD31 (platelet endothelial cell adhesion molecule [PECAM]) promoter, limiting expression to endothelial cells, monocytes, and platelets. In addition, the P-selectin sequence directs expression to secretory granules. Functional cell surface expression only occurs when the cells are activated. In a mouse model of systemic lipopolysaccharide (LPS)-induced endotoxemia, we show that expression of either anticoagulant on activated endothelium inhibits the widespread intravascular thrombosis, thrombocytopenia, and consumptive coagulopathy associated with endotoxemia. Importantly, non- LPS-treated transgenic mice had normal baseline bleeding times. We speculate that targeted delivery of anticoagulants to the endothelium may be a strategy worth pursuing in clinical sepsis to improve efficacy of systemic anticoagulation while minimizing potential hemorrhagic side effects.
Chen
Daxin
D
Department of Immunology, Imperial College London, Hammersmith Hospital, Du Cane Rd, London W12 0NN.
Giannopoulos
Konstantinos
K
Shiels
Paul G
PG
Webster
Zoe
Z
McVey
John H
JH
Kemball-Cook
Geoff
G
Tuddenham
Edward
E
Moore
Marilyn
M
Lechler
Robert
R
Dorling
Anthony
A
D000818:Animals; D001853:Bone Marrow / Q000502:physiology; D004195:Disease Models, Animal; D019446:Endotoxemia / Q000150:complications / Q000473:pathology / Q000628:therapy*; D015316:Genetic Therapy / Q000379:methods*; D006629:Hirudins / Q000235:genetics*; D006801:Humans; D007865:Leeches; D008074:Lipoproteins / Q000235:genetics*; D051379:Mice; D008822:Mice, Transgenic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D013927:Thrombosis / Q000209:etiology / Q000473:pathology / Q000628:therapy*
D006629:Hirudins; D008074:Lipoproteins; D011993:Recombinant Fusion Proteins; C051928:lipoprotein-associated coagulation inhibitor