title: β1-Na(+),K(+)-ATPase gene therapy upregulates tight junctions to rescue lipopolysaccharide-induced acute lung injury.
abstract: Acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) are associated with diverse disorders and characterized by disruption of the alveolar-capillary barrier, leakage of edema fluid into the lung, and substantial inflammation leading to acute respiratory failure. Gene therapy is a potentially powerful approach to treat ALI/ARDS through repair of alveolar epithelial function. Herein, we show that delivery of a plasmid expressing β1-subunit of the Na(+),K(+)-ATPase (β1-Na(+),K(+)-ATPase) alone or in combination with epithelial sodium channel (ENaC) α1-subunit using electroporation not only protected from subsequent lipopolysaccharide (LPS)-mediated lung injury, but also treated injured lungs. However, transfer of α1-subunit of ENaC (α1-ENaC) alone only provided protection benefit rather than treatment benefit although alveolar fluid clearance had been remarkably enhanced. Gene transfer of β1-Na(+),K(+)-ATPase, but not α1-ENaC, not only enhanced expression of tight junction protein zona occludins-1 (ZO-1) and occludin both in cultured cells and in mouse lungs, but also reduced pre-existing increase of lung permeability in vivo. These results demonstrate that gene transfer of β1-Na(+),K(+)-ATPase upregulates tight junction formation and therefore treats lungs with existing injury, whereas delivery of α1-ENaC only maintains pre-existing tight junction but not for generation. This indicates that the restoration of epithelial/endothelial barrier function may provide better treatment of ALI/ARDS.
authors.affiliation: Department of Pediatrics, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA.
authors.forename: X
authors.initials: X
authors.lastname: Lin
authors.affiliation: Department of Pediatrics, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA.
authors.forename: M
authors.initials: M
authors.lastname: Barravecchia
authors.affiliation: Department of Pediatrics, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA.
authors.forename: P
authors.initials: P
authors.lastname: Kothari
authors.affiliation: Department of Pediatrics, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA.
authors.forename: J L
authors.initials: JL
authors.lastname: Young
authors.affiliation: Department of Pediatrics, School of Medicine and Dentistry, University of Rochester, Rochester, NY, USA.
authors.forename: D A
authors.initials: DA
authors.lastname: Dean
mesh_terms: D055371:Acute Lung Injury / Q000139:chemically induced / Q000201:enzymology / Q000235:genetics* / Q000628:therapy*; D000818:Animals; D004195:Disease Models, Animal; D018274:Electroporation / Q000379:methods; D053503:Epithelial Sodium Channels / Q000627:therapeutic use; D015316:Genetic Therapy / Q000379:methods*; D008070:Lipopolysaccharides; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D012128:Respiratory Distress Syndrome / Q000628:therapy; D000254:Sodium-Potassium-Exchanging ATPase / Q000008:administration & dosage / Q000235:genetics*; D019108:Tight Junctions / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D053503:Epithelial Sodium Channels; D008070:Lipopolysaccharides; D000254:Sodium-Potassium-Exchanging ATPase