title: Epithelial organic cation transporters ensure pH-dependent drug absorption in the airway.
abstract: Most inhaled beta(2)-adrenergic agonist and anticholinergic bronchodilators have low lipid solubility because of their transient or permanent positive net charge at physiologic pH. Airway absorption of these cationic drugs is incompletely understood. We examined carrier-mediated mechanisms of cationic drug uptake by human airway epithelia. Airway tissues and epithelial cells, obtained from lung donors without preexisting lung disease, were evaluated for organic cation transporter expression by quantitative RT-PCR and immunofluorescence. For in vitro functional studies on primary airway epithelial cells, uptake of the cationic fluorophore 4-[4-(dimethylamino)-styryl]-N-methylpyridinium (ASP+) was characterized. Quantitative RT-PCR analysis demonstrated high mRNA levels for two polyspecific organic cation/carnitine transporters, OCTN1 and OCTN2, in human airway epithelia. Immunofluorescence of human airway sections confirmed OCTN1/2 protein expression, with a predominant localization to the apical portion of epithelial cells. Primary airway epithelial cells showed a carrier-mediated, temperature-sensitive and saturable uptake of ASP(+). Seventy-five to eighty percent of ASP(+) uptake was inhibited by L-carnitine, an OCTN2-carried zwitterion. The uptake was pH dependent, with approximately 3-fold lower rates at acidic (pH 5.7) than at alkaline (pH 8.2) extracellular pH. Albuterol and formoterol inhibited ASP(+) uptake, suggesting that all these molecules are carried by the same transport mechanism. These findings demonstrate the existence and functional role of a pH-dependent organic cation uptake machinery, namely OCTN1 and OCTN2, in human airway epithelia. We suggest that epithelial OCTN1/2 are involved in the delivery of inhaled cationic bronchodilators to the airway tissue.
authors.affiliation: Division of Pulmonary and Critical Care Medicine, University of Miami Miler School of Medicine, Miami, FL 33101, USA. ghorvath.mail@gmail.com
authors.forename: Gabor
authors.initials: G
authors.lastname: Horvath
authors.forename: Nathalie
authors.initials: N
authors.lastname: Schmid
authors.forename: Miryam A
authors.initials: MA
authors.lastname: Fragoso
authors.forename: Andreas
authors.initials: A
authors.lastname: Schmid
authors.forename: Gregory E
authors.initials: GE
authors.lastname: Conner
authors.forename: Matthias
authors.initials: M
authors.lastname: Salathe
authors.forename: Adam
authors.initials: A
authors.lastname: Wanner
mesh_terms: D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000420:Albuterol / Q000008:administration & dosage; D001693:Biological Transport, Active / Q000187:drug effects; D001993:Bronchodilator Agents / Q000494:pharmacology; D002331:Carnitine / Q000494:pharmacology; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000378:metabolism*; D004983:Ethanolamines / Q000008:administration & dosage; D000068759:Formoterol Fumarate; D006801:Humans; D006863:Hydrogen-Ion Concentration; D027701:Organic Cation Transport Proteins / Q000378:metabolism*; D011726:Pyridinium Compounds / Q000493:pharmacokinetics*; D012333:RNA, Messenger / Q000378:metabolism; D020545:Respiratory Mucosa / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000074058:Solute Carrier Family 22 Member 5; D027981:Symporters
chemical_list: C032315:4-(4-dimethylaminostyryl)-1-methylpyridinium; D000318:Adrenergic beta-Agonists; D001993:Bronchodilator Agents; D004983:Ethanolamines; D027701:Organic Cation Transport Proteins; D011726:Pyridinium Compounds; D012333:RNA, Messenger; C109807:SLC22A4 protein, human; C492020:SLC22A5 protein, human; D000074058:Solute Carrier Family 22 Member 5; D027981:Symporters; D000420:Albuterol; D002331:Carnitine; D000068759:Formoterol Fumarate