title: Heme Oxygenase-1 Gene Therapy Provides Cardioprotection Via Control of Post-Ischemic Inflammation: An Experimental Study in a Pre-Clinical Pig Model.
abstract: BACKGROUND
  Heme oxygenase-1 (HO-1) is an inducible stress-responsive enzyme converting heme to bilirubin, carbon monoxide, and free iron, which exerts anti-inflammatory and antiapoptotic effects. Although efficient cardioprotection after HO-1 overexpression has been reported in rodents, its role in attenuating post-ischemic inflammation is unclear.
  
  
  OBJECTIVE
  This study assessed the efficacy of recombinant adenoassociated virus (rAAV)-encoding human heme oxygenase-1 (hHO-1) in attenuating post-ischemic inflammation in a murine and a porcine ischemia/reperfusion model.
  
  
  METHODS
  Murine ischemia was induced by 45 min of left anterior descending occlusion, followed by 24 h of reperfusion and functional as well as fluorescent-activated cell sorting analysis. Porcine hearts were subjected to 60 min of ischemia and 24h of reperfusion before hemodynamic and histologic analyses were performed.
  
  
  RESULTS
  Human microvascular endothelial cells transfected with hHO-1 displayed an attenuated interleukin-6 and intercellular adhesion molecule 1 expression, resulting in reduced monocytic THP-1 cell recruitment in vitro. In murine left anterior descending occlusion and reperfusion, the post-ischemic influx of CD45(+) leukocytes, Ly-6G(+) neutrophils, and Ly-6C(high) monocytes was further exacerbated in HO-1-deficient hearts and reversed by rAAV.hHO-1 treatment. Conversely, in our porcine model of ischemia, the post-ischemic influx of myeloperoxidase-positive neutrophils and CD14(+) monocytes was reduced by 49% and 87% after rAAV.hHO-1 transduction, similar to hHO-1 transgenic pigs. Functionally, rAAV.hHO-1 and hHO-1 transgenic left ventricles displayed a smaller loss of ejection fraction than control animals.
  
  
  CONCLUSIONS
  Whereas HO-1 deficiency exacerbates post-ischemic cardiac inflammation in mice, hHO-1 gene therapy attenuates inflammation after ischemia and reperfusion in murine and porcine hearts. Regional hHO-1 gene therapy provides cardioprotection in a pre-clinical porcine ischemia/reperfusion model.
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany; Institute for Cardiovascular Prevention, Ludwig Maximillian University, Munich, Germany; Medizinische Klinik I, Klinikum Rechts der Isar, Technical University of Munich, and German Center for Cardiovascular Research, partner site Munich Heart Alliance, Munich, Germany.
authors.forename: Rabea
authors.initials: R
authors.lastname: Hinkel
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Lange
authors.affiliation: Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Petersen
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Elena
authors.initials: E
authors.lastname: Gottlieb
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany; Medizinische Klinik I, Klinikum Rechts der Isar, Technical University of Munich, and German Center for Cardiovascular Research, partner site Munich Heart Alliance, Munich, Germany.
authors.forename: Judy King Man
authors.initials: JK
authors.lastname: Ng
authors.affiliation: Department of Medicine 2, Center for Thrombosis and Hemostasis Mainz and German Center for Cardiovascular Research, partner site Rhine Main, Mainz, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Finger
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Horstkotte
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Seungmin
authors.initials: S
authors.lastname: Lee
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Thormann
authors.affiliation: Department of Medicine 2, Center for Thrombosis and Hemostasis Mainz and German Center for Cardiovascular Research, partner site Rhine Main, Mainz, Germany.
authors.forename: Maike
authors.initials: M
authors.lastname: Knorr
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Chiraz
authors.initials: C
authors.lastname: El-Aouni
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Boekstegers
authors.affiliation: Walter-Brendel-Centre for Experimental Medicine, Munich, Germany.
authors.forename: Bruno
authors.initials: B
authors.lastname: Reichart
authors.affiliation: Department of Medicine 2, Center for Thrombosis and Hemostasis Mainz and German Center for Cardiovascular Research, partner site Rhine Main, Mainz, Germany.
authors.forename: Philip
authors.initials: P
authors.lastname: Wenzel
authors.affiliation: Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut, Mariensee, Germany.
authors.forename: Heiner
authors.initials: H
authors.lastname: Niemann
authors.affiliation: Medizinische Klinik I, Klinikum Grosshadern, Ludwig Maximilian University, Munich, Germany; Medizinische Klinik I, Klinikum Rechts der Isar, Technical University of Munich, and German Center for Cardiovascular Research, partner site Munich Heart Alliance, Munich, Germany; Walter-Brendel-Centre for Experimental Medicine, Munich, Germany. Electronic address: christian.kupatt@tum.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Kupatt
mesh_terms: D000818:Animals; D000229:Dependovirus; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D051547:Heme Oxygenase-1 / Q000235:genetics*; D007249:Inflammation / Q000517:prevention & control*; D051379:Mice; D017202:Myocardial Ischemia / Q000150:complications*; D015427:Reperfusion Injury; D013552:Swine
chemical_list: C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1
keywords: adenoassociated virus; cardiomyocyte; endothelial; reperfusion; transgenic