title: Targeted omics analyses, and metabolic enzyme activity assays demonstrate maintenance of key mucociliary characteristics in long term cultures of reconstituted human airway epithelia.
abstract: 3D reconstituted respiratory epithelia have emerged as better in vitro models for toxicological testing compared to cell lines due to the conservation of key morphological features and functions. MucilAir™ is a commercially available human airway epithelia system that can potentially maintain functional attributes for up to a year, however, detailed mucociliary characteristics and xenobiotic metabolism relevant to inhaled pro-toxicant bioactivation is lacking. Here, we assessed in MucilAir™ some key biomarkers that are characteristic of the respiratory epithelia including morphology, function and xenobiotics metabolism. The end points that were measured included targeted proteomics using a panel of 243 airway surface liquid (ASL) proteins, cilia beat frequency (CBF), a qRT-PCR screen of xenobiotic metabolizing enzymes, and CYP2A6/13, CYP1A1/1B1 activity. Comparison of ASL proteomics with human sputum identified key proteins common to both matrices, but present at different levels. Xenobiotic metabolism gene profiling demonstrated strong similarities with the normal human lung and did not reveal any consistent changes when assessed over a 6 month period. Inducibility and activity of CYP1A1/1B1 and activity of CYP2A6/2A13 were present at one month in culture and maintained in one tested MucilAir™ donor for several months. In conclusion, MucilAir™ presented important morphological and metabolic characteristics of a mucociliary epithelium in short and long term culture. MucilAir™ is therefore a potentially useful model to test repeated sub-cytotoxic doses of toxicants.
authors: BAT, Group Research and Development, Regents Park Road, Southampton SO15 8TL, UK.
authors: Andrew
authors: A
authors: Baxter
authors: BAT, Group Research and Development Cambridge, Milton Road, Cambridge CB4 0WA, UK.
authors: Simon
authors: S
authors: Thain
authors: BAT, Group Research and Development, Regents Park Road, Southampton SO15 8TL, UK.
authors: Anisha
authors: A
authors: Banerjee
authors: BAT, Group Research and Development, Regents Park Road, Southampton SO15 8TL, UK.
authors: Linsey
authors: L
authors: Haswell
authors: BAT, Group Research and Development, Regents Park Road, Southampton SO15 8TL, UK.
authors: Aleesha
authors: A
authors: Parmar
authors: BAT, Group Research and Development, Regents Park Road, Southampton SO15 8TL, UK.
authors: Gary
authors: G
authors: Phillips
authors: BAT, Group Research and Development, Regents Park Road, Southampton SO15 8TL, UK. Electronic address: emmanuel_minet@BAT.com.
authors: Emmanuel
authors: E
authors: Minet
mesh_terms: D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002923:Cilia / Q000502:physiology; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D040901:Proteomics; D020545:Respiratory Mucosa / Q000378:metabolism*; D013183:Sputum / Q000378:metabolism; D018675:Toxicity Tests / Q000379:methods
chemical_list: D003577:Cytochrome P-450 Enzyme System
keywords: Air liquid interface (ALI) culture; Gene expression; Metabolism; MucilAir™; Protein expression; Respiratory epithelium