title: Evaluation of air-interfaced Calu-3 cell layers for investigation of inhaled drug interactions with organic cation transporters in vitro.
abstract: A physiologically pertinent in vitro model is urgently needed for probing interactions between inhaled drugs and the organic cation transporters (OCT) in the bronchial epithelium. This study evaluated OCT expression, functionality, inhibition by common inhaled drugs and impact on formoterol transepithelial transport in layers of human bronchial epithelial Calu-3 cells grown at an air-liquid interface. 21 day old Calu-3 layers expressed OCT1, OCT3, OCTN1 and OCTN2 whereas OCT2 could not be detected. Quantification of the cellular uptake of the OCT substrate ASP(+) in presence of inhibitors suggested several OCT were functional at the apical side of the cell layers. ASP(+) uptake was reduced by the bronchodilators formoterol, salbutamol (albuterol), ipratropium and the glucocorticoid budesonide. However, the OCT inhibitory properties of the two β(2)-mimetics were suppressed at therapeutically relevant concentrations. The absorptive permeability of formoterol across the cell layers was enhanced at a high drug concentration shown to decrease ASP(+) uptake by ∼50% as well as in presence of the OCT inhibitor tetraethylammonium (TEA). Secretory transport was unaffected by the drug concentration but was reduced by TEA. Our data indicate air-interfaced Calu-3 layers offer a low-cost in vitro model suitable for assessing inhaled drug-OCT interactions in the bronchial epithelium.
authors.affiliation: Division of Drug Delivery and Tissue Engineering, School of Pharmacy, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom.
authors.forename: Manali
authors.initials: M
authors.lastname: Mukherjee
authors.affiliation: Immune Modulation Research Group, School of Pharmacy, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom.
authors.forename: D I
authors.initials: DI
authors.lastname: Pritchard
authors.affiliation: Division of Drug Delivery and Tissue Engineering, School of Pharmacy, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom. Electronic address: cynthia.bosquillon@nottingham.ac.uk.
authors.forename: C
authors.initials: C
authors.lastname: Bosquillon
mesh_terms: D000280:Administration, Inhalation; D000420:Albuterol / Q000494:pharmacology; D001692:Biological Transport; D001993:Bronchodilator Agents / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D019819:Budesonide / Q000494:pharmacology; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D004983:Ethanolamines / Q000378:metabolism / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000378:metabolism; D000068759:Formoterol Fumarate; D005786:Gene Expression Regulation; D006801:Humans; D009241:Ipratropium / Q000494:pharmacology; D027701:Organic Cation Transport Proteins / Q000037:antagonists & inhibitors / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D027702:Organic Cation Transporter 1 / Q000037:antagonists & inhibitors / Q000378:metabolism; D000075102:Organic Cation Transporter 2; D010539:Permeability; D011726:Pyridinium Compounds / Q000378:metabolism; D020545:Respiratory Mucosa / Q000187:drug effects* / Q000378:metabolism; D000074058:Solute Carrier Family 22 Member 5; D027981:Symporters; D000099254:Organic Cation Transporter 3
chemical_list: D000420:Albuterol; D001993:Bronchodilator Agents; D019819:Budesonide; D004983:Ethanolamines; D005456:Fluorescent Dyes; D000068759:Formoterol Fumarate; D009241:Ipratropium; D027701:Organic Cation Transport Proteins; D027702:Organic Cation Transporter 1; D000075102:Organic Cation Transporter 2; D011726:Pyridinium Compounds; D000074058:Solute Carrier Family 22 Member 5; D027981:Symporters; D000099254:Organic Cation Transporter 3; C491013:SLC22A2 protein, human; C109807:SLC22A4 protein, human; C492020:SLC22A5 protein, human; C052247:4-(4-diethylaminostyryl)-N-methylpyridinium