title: Inhibition of intravascular thrombosis in murine endotoxemia by targeted expression of hirudin and tissue factor pathway inhibitor analogs to activated endothelium.
abstract: 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.
authors.affiliation: Department of Immunology, Imperial College London, Hammersmith Hospital, Du Cane Rd, London W12 0NN.
authors.forename: Daxin
authors.initials: D
authors.lastname: Chen
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Giannopoulos
authors.forename: Paul G
authors.initials: PG
authors.lastname: Shiels
authors.forename: Zoe
authors.initials: Z
authors.lastname: Webster
authors.forename: John H
authors.initials: JH
authors.lastname: McVey
authors.forename: Geoff
authors.initials: G
authors.lastname: Kemball-Cook
authors.forename: Edward
authors.initials: E
authors.lastname: Tuddenham
authors.forename: Marilyn
authors.initials: M
authors.lastname: Moore
authors.forename: Robert
authors.initials: R
authors.lastname: Lechler
authors.forename: Anthony
authors.initials: A
authors.lastname: Dorling
mesh_terms: 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*
chemical_list: D006629:Hirudins; D008074:Lipoproteins; D011993:Recombinant Fusion Proteins; C051928:lipoprotein-associated coagulation inhibitor