title: Post-ischaemic silencing of p66Shc reduces ischaemia/reperfusion brain injury and its expression correlates to clinical outcome in stroke.
abstract: OBJECTIVE
  Constitutive genetic deletion of the adaptor protein p66(Shc) was shown to protect from ischaemia/reperfusion injury. Here, we aimed at understanding the molecular mechanisms underlying this effect in stroke and studied p66(Shc) gene regulation in human ischaemic stroke.
  
  
  RESULTS
  Ischaemia/reperfusion brain injury was induced by performing a transient middle cerebral artery occlusion surgery on wild-type mice. After the ischaemic episode and upon reperfusion, small interfering RNA targeting p66(Shc) was injected intravenously. We observed that post-ischaemic p66(Shc) knockdown preserved blood-brain barrier integrity that resulted in improved stroke outcome, as identified by smaller lesion volumes, decreased neurological deficits, and increased survival. Experiments on primary human brain microvascular endothelial cells demonstrated that silencing of the adaptor protein p66(Shc) preserves claudin-5 protein levels during hypoxia/reoxygenation by reducing nicotinamide adenine dinucleotide phosphate oxidase activity and reactive oxygen species production. Further, we found that in peripheral blood monocytes of acute ischaemic stroke patients p66(Shc) gene expression is transiently increased and that this increase correlates with short-term neurological outcome.
  
  
  CONCLUSIONS
  Post-ischaemic silencing of p66(Shc) upon reperfusion improves stroke outcome in mice while the expression of p66(Shc) gene correlates with short-term outcome in patients with ischaemic stroke.
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: R D
authors.initials: RD
authors.lastname: Spescha
authors.affiliation: Institute for Biomedical Engineering, Swiss Federal Institute of Technology Zurich (ETHZ), Zurich, Switzerland.
authors.forename: J
authors.initials: J
authors.lastname: Klohs
authors.affiliation: Department of Neurology, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Semerano
authors.affiliation: Department of Neurology, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Giacalone
authors.affiliation: Division of Psychiatry Research, University of Zurich, Schlieren, Switzerland.
authors.forename: R S
authors.initials: RS
authors.lastname: Derungs
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: M F
authors.initials: MF
authors.lastname: Reiner
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland.
authors.forename: D
authors.initials: D
authors.lastname: Rodriguez Gutierrez
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland.
authors.forename: N
authors.initials: N
authors.lastname: Mendez-Carmona
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Glanzmann
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: G
authors.initials: G
authors.lastname: Savarese
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland Department of Cardiology, Charité - Universitätsmedizin Berlin, Campus Benjamin Franklin, Berlin, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Kränkel
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: A
authors.initials: A
authors.lastname: Akhmedov
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: S
authors.initials: S
authors.lastname: Keller
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland.
authors.forename: P
authors.initials: P
authors.lastname: Mocharla
authors.affiliation: Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland Institute of Physiology, University of Zurich, Zurich, Switzerland.
authors.forename: M R
authors.initials: MR
authors.lastname: Kaufmann
authors.affiliation: Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland Institute of Physiology, University of Zurich, Zurich, Switzerland.
authors.forename: R H
authors.initials: RH
authors.lastname: Wenger
authors.affiliation: Institute of Veterinary Physiology, University of Zurich, Zurich, Switzerland.
authors.forename: J
authors.initials: J
authors.lastname: Vogel
authors.affiliation: Division of Psychiatry Research, University of Zurich, Schlieren, Switzerland.
authors.forename: L
authors.initials: L
authors.lastname: Kulic
authors.affiliation: Division of Psychiatry Research, University of Zurich, Schlieren, Switzerland.
authors.forename: R M
authors.initials: RM
authors.lastname: Nitsch
authors.affiliation: Department of Internal Medicine, Cantonal Hospital of Baden, Baden, Switzerland.
authors.forename: J H
authors.initials: JH
authors.lastname: Beer
authors.affiliation: Department of Neurology, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: L
authors.initials: L
authors.lastname: Peruzzotti-Jametti
authors.affiliation: Department of Neurology, San Raffaele Scientific Institute, Milan, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Sessa
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland Cardiology, University Heart Center, University Hospital, Zurich, Switzerland.
authors.forename: T F
authors.initials: TF
authors.lastname: Lüscher
authors.affiliation: Center for Molecular Cardiology, University of Zurich, Wagistrasse 12, Schlieren CH-8952, Switzerland Zurich Center for Integrative Human Physiology (ZIHP), University of Zurich, Zurich, Switzerland giovanni.camici@uzh.ch giovannicamici@hotmail.com.
authors.forename: G G
authors.initials: GG
authors.lastname: Camici
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D000704:Analysis of Variance; D000818:Animals; D001812:Blood-Brain Barrier / Q000502:physiology; D001930:Brain Injuries / Q000517:prevention & control*; D016022:Case-Control Studies; D002478:Cells, Cultured; D062507:Claudin-5 / Q000187:drug effects; D042783:Endothelial Cells / Q000502:physiology; D005260:Female; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D020868:Gene Silencing / Q000502:physiology*; D006801:Humans; D020244:Infarction, Middle Cerebral Artery; D057775:Ischemic Postconditioning / Q000379:methods; D008297:Male; D008810:Mice, Inbred C57BL; D008833:Microcirculation / Q000502:physiology; D008875:Middle Aged; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000494:pharmacology; D015427:Reperfusion Injury / Q000517:prevention & control*; D055765:Shc Signaling Adaptor Proteins / Q000235:genetics* / Q000502:physiology; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D020521:Stroke / Q000517:prevention & control*; D016896:Treatment Outcome
chemical_list: D062507:Claudin-5; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; C528024:SHC1 protein, human; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1
keywords: Endothelium; Free radicals; Ischaemia; Reperfusion; Stroke