title: The prediction of drug metabolism using scaffold-mediated enhancement of the induced cytochrome P450 activities in fibroblasts by hepatic transcriptional regulators.
abstract: A reliable, reproducible, and convenient in vitro platform for drug metabolism determination and toxicity prediction is of tremendous value but still lacking. In the present study, a collection of 24 hepatic transcription factors and nuclear receptors in different combinations were surveyed, and 10 among them were finally selected to induce the expression and enzyme activities of cytochrome P450 (CYP) 3A4, 1B1, and 2C9 in human dermal fibroblasts (HDFs). The expression and activities of these CYPs in the induced HDFs were higher than those in commonly used hepatoma cell lines. High CYP expression and activities could be further enhanced by culturing the induced HDFs either as spheroids or into several kinds of scaffolds, particularly the tri-copolymer scaffold composed of gelatin, chondroitin and hyaluronan. More strikingly, there showed a synergistic effect of seeding and culturing the spheroids into the tri-copolymer scaffold. Scanning electron microscopy and confocal microscopy disclosed well accommodation of these spheroids inside the scaffolds and displayed a high survival rate. Moreover, the spheroid/scaffold constructs could metabolize an anti-hypertension drug nifedipine into oxidized nifedipine, showing their applicability in studying drug metabolism. This study presents a strategy to induce the expression and enzyme activities of critical CYPs in HDFs, and may have potential to establish an in vitro platform to study drug metabolism and to predict the possible human risk of drug toxicity.
authors.affiliation: Institute of Biotechnology, College of Bioresources and Agriculture, National Taiwan University, 4F, No. 81, Chang-Xing St., Taipei 106, Taiwan. d97642003@ntu.edu.tw
authors.forename: Tsai-Shin
authors.initials: TS
authors.lastname: Chiang
authors.forename: Kai-Chiang
authors.initials: KC
authors.lastname: Yang
authors.forename: Shu-Kai
authors.initials: SK
authors.lastname: Zheng
authors.forename: Ling-Ling
authors.initials: LL
authors.lastname: Chiou
authors.forename: Wen-Ming
authors.initials: WM
authors.lastname: Hsu
authors.forename: Feng-Huei
authors.initials: FH
authors.lastname: Lin
authors.forename: Guan-Tarn
authors.initials: GT
authors.lastname: Huang
authors.forename: Hsuan-Shu
authors.initials: HS
authors.lastname: Lee
mesh_terms: D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D020405:Dermis / Q000166:cytology; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D008658:Inactivation, Metabolic; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011108:Polymers / Q000494:pharmacology; D018874:Spheroids, Cellular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D054457:Tissue Scaffolds / Q000737:chemistry*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D011108:Polymers; D014157:Transcription Factors; D003577:Cytochrome P-450 Enzyme System