title: E-selectin inhibition with GMI-1271 decreases venous thrombosis without profoundly affecting tail vein bleeding in a mouse model.
abstract: Selectins, such as E-selectin (CD62E), function in venous thrombosis by binding and activating immune cells to initiate the coagulation cascade. GMI-1271 is a small molecule antagonist that inhibits E-selectin activity. Here we determine whether inhibition of E-selectin is sufficient to decrease acute venous thrombosis and associated inflammatory events in both prophylactic and treatment protocols without significantly affecting haemostasis. Male C57BL/6 mice underwent surgery for experimental thrombosis induction and were harvested at peak thrombus formation in our animal model, two days post induction. Groups included non-thrombosed true controls, shams, controls, and prophylactic or treatment groups of GMI-1271 (10 mg/kg intraperitoneal BID (twice a day) and low-molecular-weight heparin (LMWH, Lovenox 6 mg/kg subcutaneously (SC), once a day (SID). Compared with control animals, prophylaxis or treatment with LMWH and GMI-1271 in a dose-dependent manner significantly decreased thrombosis. GMI-1271 significantly lowered tail bleeding times when compared to LMWH. GMI-1271 and LMWH prophylactically administered significantly decreased vein wall neutrophil cell extravasation. However, all treatment and prophylactic therapies significantly decreased vein wall monocyte extravasation versus controls. GMI-1271 prophylactic therapy significantly decreased intra-thrombus cell counts versus control animals and other treatment groups. Immunohistochemistry confirmed that both treatments with GMI-1271 and LMWH significantly decreased activated leukocyte migration. GMI-1271 therapy significantly decreased thrombus weight and resulted in significantly lower bleeding times than LMWH. GMI-1271 treated mice showed decreased local and systemic inflammatory effects while modulating neutrophil activation, suggesting that GMI-1271 is a viable therapeutic candidate for venous thrombosis prophylaxis and treatment.
authors.forename: Dorian L
authors.initials: DL
authors.lastname: Culmer
authors.forename: Misha L
authors.initials: ML
authors.lastname: Dunbar
authors.forename: Angela E
authors.initials: AE
authors.lastname: Hawley
authors.forename: Suman
authors.initials: S
authors.lastname: Sood
authors.forename: Robert E
authors.initials: RE
authors.lastname: Sigler
authors.forename: Peter K
authors.initials: PK
authors.lastname: Henke
authors.forename: Thomas W
authors.initials: TW
authors.lastname: Wakefield
authors.forename: John L
authors.initials: JL
authors.lastname: Magnani
authors.affiliation: Daniel D. Myers, Jr., DVM, MPH, DACLAM, University of Michigan, North Campus Research Complex, Building 26, Room 263N, 2800 Plymouth Road, Ann Arbor, MI 48109-2800, USA, Tel.: +1 734 763 0940, E-mail: ddmyers@umich.edu.
authors.forename: Daniel D
authors.initials: DD
authors.lastname: Myers
mesh_terms: D000818:Animals; D018395:CA-19-9 Antigen; D002465:Cell Movement; D004195:Disease Models, Animal; D019040:E-Selectin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D005732:Gangliosides / Q000737:chemistry / Q000627:therapeutic use*; D006470:Hemorrhage / Q000209:etiology / Q000517:prevention & control*; D006495:Heparin, Low-Molecular-Weight / Q000627:therapeutic use; D006801:Humans; D007249:Inflammation / Q000188:drug therapy*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018716:Molecular Mimicry; D009504:Neutrophils / Q000276:immunology*; D013623:Tail / Q000033:anatomy & histology; D014680:Veins / Q000502:physiology*; D020246:Venous Thrombosis / Q000150:complications / Q000188:drug therapy*
chemical_list: D018395:CA-19-9 Antigen; D019040:E-Selectin; D005732:Gangliosides; D006495:Heparin, Low-Molecular-Weight; C064564:sialyl Le(a) ganglioside
keywords: E-selectin inhibition; animal models; glycomimetic; inflammation; venous thrombosis