Integrated assessment of rifampicin‐mediated induction of transporters and drug‐metabolizing enzymes in a long‐term human liver tissue chip system
Year: 2026
Type: article
Abstract: Rifampicin is a prototypical clinical inducer used in drug–drug interaction (DDI) studies. However, the clinical relevance and predictability of rifampicin‐mediated hepatic transporter induction remain poorly defined. Conventional hepatocyte culture models fail to capture clinically relevant regulation of transporters and drug‐metabolizing enzymes (DMEs), particularly at the protein level. In this study, we characterized a liver tissue chip (LTC) as a new approach methodology (NAM) for long‐term transporter‐ and DME‐mediated induction risk assessment. Transporter‐certified® cryopreserved primary human hepatocytes from three donors were cultured in the LTC under recirculating flow with or without 10 μM rifampicin for 7 days. The LTC facilitated simultaneous mRNA‐protein quantification for transporters and mRNA‐protein‐activity profiling for cytochrome P450s (CYPs) within individual chips across treatment and control conditions. In addition, extracellular bile acid profiling and intracellular rifampicin concentrations were measured by LC–MS/MS. Rifampicin induced multiple transporters and enzymes at the protein level, including organic anion transporting polypeptide (OATP)2B1, P‐glycoprotein, CYP3A4, CYP2C19, and uridine 5’‐diphospho‐glucuronosyltransferase (UGT)1A4. Induction patterns differed between transporter mRNA and protein in many cases, suggesting regulatory mechanisms beyond PXR‐mediated activation. Matched mRNA‐protein‐activity measurements of five CYP enzymes also revealed discrepancies in rifampicin‐mediated induction magnitude. Rifampicin altered bile acid composition, increasing taurine‐conjugated bile acids and inducing bile acid‐related genes. Measured intracellular rifampicin concentrations differed substantially from experimentally employed media concentrations. Overall, these findings demonstrate that the LTC enables integrated assessment of mRNA, protein, and activity measurements in addition to bile acid profiles and intracellular drug exposure, which may improve mechanistic understanding of induction processes.
Source: UNC Libraries
DOI: 10.17615/fy1d-dn68
Institution: University of North Carolina at Chapel Hill
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: green
Funders: Region Syddanmark, U.S. Department of Veterans Affairs, Arthritis Foundation, American Orthopaedic Foot and Ankle Society, Natural Science Foundation of Shanghai, Grantová Agentura, Univerzita Karlova, Deutsche Bundesstiftung Umwelt, Japan Agency for Medical Research and Development, UK Research and Innovation, Ministerio de Ciencia, Innovación y Universidades, Australian Government, Seerave Foundation, Försvarsdepartementet, Sveriges, Kern Family Foundation, Department for Environment, Food and Rural Affairs, UK Government, European Commission, Deutsche Forschungsgemeinschaft, Agence Nationale de la Recherche, National Natural Science Foundation of China, Ministerstvo Školství, Mládeže a Tělovýchovy, Grantová Agentura České Republiky, ZonMw, Bundesministerium für Bildung und Forschung, Austrian Science Fund, Generalitat de Catalunya, China Postdoctoral Science Foundation, Stichting MS Research, Ministry of Health, Labour and Welfare, Guangdong Medical Research Foundation, Fondation ARC pour la Recherche sur le Cancer, Ligue Contre le Cancer, Vetenskapsrådet, Türkiye Bilimsel ve Teknolojik Araştırma Kurumu, Institut Universitaire de France, Natural Science Foundation of Chongqing, Norges Forskningsråd, Turun Yliopisto, Malmö Högskola, Institut National Du Cancer, Natural Science Foundation for Young Scientists of Shanxi Province, National Institutes of Health, Qatar National Research Fund, Horizon 2020 Framework Programme, Clinical and Translational Science Collaborative of Cleveland, School of Medicine, Case Western Reserve University, Chief Scientist Office, Scottish Government Health and Social Care Directorate, HORIZON EUROPE Framework Programme, Canadian Institutes of Health Research, Natural Sciences and Engineering Research Council of Canada, Natural Environment Research Council, Japan Society for the Promotion of Science, European Regional Development Fund, University Natural Science Research Project of Anhui Province, National Key Research and Development Program of China, Faculty of Medicine and Dentistry, University of Alberta, Basic and Applied Basic Research Foundation of Guangdong Province, Företagsforskarskolan för Samverkan och Innovation, Umeå Universitet, National Heart, Lung, and Blood Institute, H2020 European Research Council, Natural Science Foundation of Zhejiang Province, Cliff Lede Family Charitable Foundation
OpenAlex ID: W7203994269
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