title: Thrombin mutant W215A/E217A treatment improves neurological outcome and reduces cerebral infarct size in a mouse model of ischemic stroke.
abstract: OBJECTIVE
  Treatment of ischemic stroke by activation of endogenous plasminogen using tissue plasminogen activator is limited by bleeding side effects. In mice, treatment of experimental ischemic stroke with activated protein C improves outcomes; however, activated protein C also has bleeding side effects. In contrast, activation of endogenous protein C using thrombin mutant W215A/E217A (WE) is antithrombotic without hemostasis impairment in primates. Therefore, we investigated the outcome of WE-treated experimental ischemic stroke in mice.
  
  
  METHODS
  The middle cerebral artery was occluded with a filament for 60 minutes to induce ischemic stroke. Vehicle, recombinant WE, or tissue plasminogen activator was administered during middle cerebral artery occlusion or 2 hours after middle cerebral artery occlusion. Neurological performance was scored daily. Intracranial bleeding and cerebral infarct size, defined by 2,3,5-triphenyltetrazolium chloride exclusion, were determined on autopsy. Hemostasis was evaluated using tail bleeding tests.
  
  
  RESULTS
  WE improved neurological performance scores, increased laser Doppler flowmetry-monitored post-middle cerebral artery occlusion reperfusion of the parietal cortex, and reduced 2,3,5-triphenyltetrazolium chloride-defined cerebral infarct size versus vehicle controls. However, unlike tissue plasminogen activator, WE did not increase tail bleeding or intracranial hemorrhage.
  
  
  CONCLUSIONS
  WE treatment is neuroprotective without hemostasis impairment in experimental acute ischemic stroke in mice and thus may provide an alternative to tissue plasminogen activator for stroke treatment.
authors.affiliation: Department of Biomedical Engineering, Oregon Health & Science University, 3303 SW Bond Ave, Portland, OR 97239, USA.
authors.forename: Michelle A
authors.initials: MA
authors.lastname: Berny-Lang
authors.forename: Sawan
authors.initials: S
authors.lastname: Hurst
authors.forename: Erik I
authors.initials: EI
authors.lastname: Tucker
authors.forename: Leslie A
authors.initials: LA
authors.lastname: Pelc
authors.forename: Ruikang K
authors.initials: RK
authors.lastname: Wang
authors.forename: Patricia D
authors.initials: PD
authors.lastname: Hurn
authors.forename: Enrico
authors.initials: E
authors.lastname: Di Cera
authors.forename: Owen J T
authors.initials: OJ
authors.lastname: McCarty
authors.forename: András
authors.initials: A
authors.lastname: Gruber
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000627:therapeutic use*; D002545:Brain Ischemia / Q000188:drug therapy / Q000473:pathology; D002544:Cerebral Infarction / Q000188:drug therapy* / Q000473:pathology* / Q000503:physiopathology; D004195:Disease Models, Animal; D006487:Hemostasis; D006801:Humans; D020244:Infarction, Middle Cerebral Artery / Q000188:drug therapy; D017078:Laser-Doppler Flowmetry; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D020521:Stroke / Q000188:drug therapy / Q000473:pathology; D013917:Thrombin / Q000235:genetics* / Q000627:therapeutic use*; D010959:Tissue Plasminogen Activator / Q000627:therapeutic use; D016896:Treatment Outcome
chemical_list: D000925:Anticoagulants; D013917:Thrombin; D010959:Tissue Plasminogen Activator