title: Specific modulation of airway epithelial tight junctions by apical application of an occludin peptide.
abstract: Tight junctions are directly involved in regulating the passage of ions and macromolecules (gate functions) in epithelial and endothelial cells. The modulation of these gate functions to transiently regulate the paracellular permeability of large solutes and ions could increase the delivery of pharmacological agents or gene transfer vectors. To reduce the inflammatory responses caused by tight junction-regulating agents, alternative strategies directly targeting specific tight junction proteins could prove to be less toxic to airway epithelia. The apical delivery of peptides corresponding to the first extracellular loop of occludin to transiently modulate apical paracellular flux has been demonstrated in intestinal epithelia. We hypothesized that apical application of these occludin peptides could similarly modulate tight junction permeability in airway epithelia. Thus, we investigated the effects of apically applied occludin peptide on the paracellular permeability of molecular tracers and viral vectors in well differentiated human airway epithelial cells. The effects of occludin peptide on cellular toxicity, tight junction protein expression and localization, and membrane integrity were also assessed. Our data showed that apically applied occludin peptide significantly reduced transepithelial resistance in airway epithelia and altered tight junction permeability in a concentration-dependent manner. These alterations enhanced the paracellular flux of dextrans as well as gene transfer vectors. The occludin peptide redistributed occludin but did not alter the expression or distribution of ZO-1, claudin-1, or claudin-4. These data suggest that specific targeting of occludin could be a better-suited alternative strategy for tight junction modulation in airway epithelial cells compared with current agents that modulate tight junctions.
authors.affiliation: Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, USA.
authors.forename: Ruth S
authors.initials: RS
authors.lastname: Everett
authors.forename: Miriam K
authors.initials: MK
authors.lastname: Vanhook
authors.forename: Nadia
authors.initials: N
authors.lastname: Barozzi
authors.forename: Istvan
authors.initials: I
authors.lastname: Toth
authors.forename: Larry G
authors.initials: LG
authors.lastname: Johnson
mesh_terms: D000595:Amino Acid Sequence; D002478:Cells, Cultured; D062445:Claudin-1; D062506:Claudin-4; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism*; D008969:Molecular Sequence Data; D062793:Occludin; D010446:Peptide Fragments / Q000008:administration & dosage; D010539:Permeability / Q000187:drug effects; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism; D012137:Respiratory System / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019108:Tight Junctions / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D014161:Transduction, Genetic; D062826:Zonula Occludens-1 Protein
chemical_list: C568108:CLDN1 protein, human; C568157:CLDN4 protein, human; D062445:Claudin-1; D062506:Claudin-4; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; D010446:Peptide Fragments; D010750:Phosphoproteins; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; C508502:occludin peptide (90-103)