title: Xenobiotic metabolism in differentiated human bronchial epithelial cells.
abstract: Differentiated human bronchial epithelial cells in air liquid interface cultures (ALI-PBEC) represent a promising alternative for inhalation studies with rodents as these 3D airway epithelial tissue cultures recapitulate the human airway in multiple aspects, including morphology, cell type composition, gene expression and xenobiotic metabolism. We performed a detailed longitudinal gene expression analysis during the differentiation of submerged primary human bronchial epithelial cells into ALI-PBEC to assess the reproducibility and inter-individual variability of changes in transcriptional activity during this process. We generated ALI-PBEC cultures from four donors and focussed our analysis on the expression levels of 362 genes involved in biotransformation, which are of primary importance for toxicological studies. Expression of various of these genes (e.g., GSTA1, ADH1C, ALDH1A1, CYP2B6, CYP2F1, CYP4B1, CYP4X1 and CYP4Z1) was elevated following the mucociliary differentiation of airway epithelial cells into a pseudo-stratified epithelial layer. Although a substantial number of genes were differentially expressed between donors, the differences in fold changes were generally small. Metabolic activity measurements applying a variety of different cytochrome p450 substrates indicated that epithelial cultures at the early stages of differentiation are incapable of biotransformation. In contrast, mature ALI-PBEC cultures were proficient in the metabolic conversion of a variety of substrates albeit with considerable variation between donors. In summary, our data indicate a distinct increase in biotransformation capacity during differentiation of PBECs at the air-liquid interface and that the generation of biotransformation competent ALI-PBEC cultures is a reproducible process with little variability between cultures derived from four different donors.
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Postal Zone S4-P, PO Box 9600, 2300 RC, Leiden, The Netherlands.
authors.forename: Jan J W A
authors.initials: JJWA
authors.lastname: Boei
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Postal Zone S4-P, PO Box 9600, 2300 RC, Leiden, The Netherlands.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Vermeulen
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Postal Zone S4-P, PO Box 9600, 2300 RC, Leiden, The Netherlands.
authors.forename: Binie
authors.initials: B
authors.lastname: Klein
authors.affiliation: Department of Pulmonology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
authors.forename: Pieter S
authors.initials: PS
authors.lastname: Hiemstra
authors.affiliation: Department of Pulmonology, Leiden University Medical Center, 2300 RC, Leiden, The Netherlands.
authors.forename: Renate M
authors.initials: RM
authors.lastname: Verhoosel
authors.affiliation: Department of Toxicogenomics, Maastricht University, Maastricht, The Netherlands.
authors.forename: Danyel G J
authors.initials: DGJ
authors.lastname: Jennen
authors.affiliation: Unidad de Hepatología Experimental, Instituto de Investigación Sanitaria-Fundación Hospital La Fe, 46009, Valencia, Spain.
authors.forename: Agustin
authors.initials: A
authors.lastname: Lahoz
authors.affiliation: Genedata AG, 4053, Basel, Switzerland.
authors.forename: Hans
authors.initials: H
authors.lastname: Gmuender
authors.affiliation: Department of Human Genetics, Leiden University Medical Center, Postal Zone S4-P, PO Box 9600, 2300 RC, Leiden, The Netherlands. h.vrieling@lumc.nl.
authors.forename: Harry
authors.identifier: http://orcid.org/0000-0002-3034-2664
authors.initials: H
authors.lastname: Vrieling
mesh_terms: D001551:Benz(a)Anthracenes / Q000493:pharmacokinetics; D001564:Benzo(a)pyrene / Q000493:pharmacokinetics; D001711:Biotransformation / Q000235:genetics; D001980:Bronchi / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D003580:Cytochromes / Q000235:genetics / Q000378:metabolism; D004798:Enzymes / Q000235:genetics; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D000072317:Polychlorinated Dibenzodioxins / Q000493:pharmacokinetics; D015203:Reproducibility of Results; D015262:Xenobiotics / Q000378:metabolism / Q000493:pharmacokinetics*
chemical_list: D001551:Benz(a)Anthracenes; D003580:Cytochromes; D004798:Enzymes; D000072317:Polychlorinated Dibenzodioxins; D015262:Xenobiotics; D001564:Benzo(a)pyrene; C013464:cytochrome P420; C026486:1,2,5,6-dibenzanthracene
keywords: Bronchial epithelial cells; Cytochrome P450; Gene expression profiling; Lung; Metabolic activity