title: Electroporation-mediated gene delivery of Na+,K+ -ATPase, and ENaC subunits to the lung attenuates acute respiratory distress syndrome in a two-hit porcine model.
abstract: BACKGROUND
  Acute respiratory distress syndrome (ARDS) is a common cause of organ failure with an associated mortality rate of 40%. The initiating event is disruption of alveolar-capillary interface causing leakage of edema into alveoli.
  
  
  OBJECTIVE
  Electroporation-mediated gene delivery of epithelial sodium channel (ENaC) and Na+,K+ -ATPase into alveolar cells would improve alveolar clearance of edema and attenuate ARDS.
  
  
  METHODS
  Pigs were anesthetized and instrumented, and the superior mesenteric artery was clamped to cause gut ischemia/reperfusion injury and peritoneal sepsis by fecal clot implantation. Animals were ventilated according to ARDSnet protocol. Four hours after injury, animals were randomized into groups: (i) treatment: Na+,K+ -ATPase/ENaC plasmid (n = 5) and (ii) control: empty plasmid (n = 5). Plasmids were delivered to the lung using bronchoscope. Electroporation was delivered using eight-square-wave electric pulses across the chest. Following electroporation, pigs were monitored 48 h.
  
  
  RESULTS
  The Pao2/Fio2 ratio and lung compliance were higher in the treatment group. Lung wet/dry ratio was lower in the treatment group. Relative expression of the Na+,K+ -ATPase transgene was higher throughout lungs receiving treatment plasmids. Quantitative histopathology revealed a reduction in intra-alveolar fibrin in the treatment group. Bronchoalveolar lavage showed increased surfactant protein B in the treatment group. Survival was improved in the treatment group.
  
  
  CONCLUSIONS
  Electroporation-mediated transfer of Na+,K+ -ATPase/ENaC plasmids improved lung function, reduced fibrin deposits, decreased lung edema, and improved survival in a translational porcine model of ARDS. Gene therapy can attenuate ARDS pathophysiology in a high-fidelity animal model, suggesting a potential new therapy for patients.
authors.affiliation: *Department of Surgery, SUNY Upstate Medical University, Syracuse; †Department of Biology, SUNY Cortland, Cortland; and Departments of ‡Pediatrics and §Biomedical Engineering, University of Rochester, Rochester, New York.
authors.forename: Bryanna M
authors.initials: BM
authors.lastname: Emr
authors.forename: Shreyas
authors.initials: S
authors.lastname: Roy
authors.forename: Michaela
authors.initials: M
authors.lastname: Kollisch-Singule
authors.forename: Louis A
authors.initials: LA
authors.lastname: Gatto
authors.forename: Michael
authors.initials: M
authors.lastname: Barravecchia
authors.forename: Xin
authors.initials: X
authors.lastname: Lin
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Young
authors.forename: Guirong
authors.initials: G
authors.lastname: Wang
authors.forename: Jiao
authors.initials: J
authors.lastname: Liu
authors.forename: Joshua
authors.initials: J
authors.lastname: Satalin
authors.forename: Kathleen
authors.initials: K
authors.lastname: Snyder
authors.forename: Gary F
authors.initials: GF
authors.lastname: Nieman
authors.forename: David A
authors.initials: DA
authors.lastname: Dean
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D018274:Electroporation / Q000379:methods*; D053503:Epithelial Sodium Channels / Q000096:biosynthesis* / Q000235:genetics; D018014:Gene Transfer Techniques; D008168:Lung / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D010957:Plasmids / Q000235:genetics / Q000494:pharmacology*; D012128:Respiratory Distress Syndrome / Q000235:genetics / Q000473:pathology / Q000503:physiopathology / Q000628:therapy; D013552:Swine
chemical_list: D053503:Epithelial Sodium Channels