Cross-species comparison of CAR-mediated procarcinogenic key events in a 3D liver microtissue model
Open accessAbstract
Characterisation of the mode of action (MOA) of constitutive androstane receptor (CAR)-mediated rodent liver tumours involves measurement 5 key events including activation of the CAR receptor, altered gene expression, hepatocellular proliferation, clonal expansion and increased hepatocellular adenomas/carcinomas. To test whether or not liver 3D microtissues (LiMTs) recapitulate CAR- mediated procarcinogenic key events in response to the prototypical CAR activator phenobarbital (PB) we performed hepatocyte proliferation (LI%) analysis in rat and human LiMTs using a microTMA technology in conjunction with integrated transcriptomics (microarray) and proteomics analysis. The rationale for this approach was that LiMTs containing parenchymal and non-parenchymal cells (NPCs) are more physiologically representative of liver and thus would generate data more relevant to the in vivo situation. Rat and human LiMTs were treated with PB over a range of concentrations (500 uM - 2000 uM) and times (24 h - 96 h) in a dose-response/time-course analysis. There was a dose-dependent induction of LI% in rat LiMTs, however there was little or no effect of PB on LI% in human LiMTs. ATP levels in the rat and human LiMTs were similar to control in all of the PB treatments. There was also a dose- and time-dependent PB-mediated RNA induction of CAR regulated genes CYP2B6/Cyp2b2, CYP3A7/Cyp3a9 and UGT1A6/Ugt1a6 in human and rat LiMTs, respectively. These CAR regulated genes were also upregulated at the protein level. Ingenuity pathways analysis (IPA) indicated that there was a significant (Z score >2.0;-log p value >) activation of CAR by PB in both human and rat LiMTs. These results indicate that human and rat LiMTs showed the expected responses at the level of PB-induced hepatocyte proliferation and enzyme induction with rat LiMTs showing significant dose-dependent effects while human LiMTs showed no proliferation response but did show dose-dependent enzyme induction at the RNA and protein levels. In conclusion LiMTs serve as a model to provide mechanistic data for 3 of the 5 key events considered necessary to establish a CAR-mediated MOA for liver tumourigenesis and thus can potentially reduce the use of animals when compiling mechanistic data packages.
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The effects of the CAR-driven c-Myc-Foxm1 axis are not recapitulated in humanized CAR transgenic mice or human hepatocytes.
“these effects are not recapitulated in humanized CAR transgenic mice or human hepatocytes,45, 46, 47”
From Constitutive Androstane Receptor Regulates Bile Acid Homeostasis and Metabolism During Liver Regeneration (2026) · cites this paper as [47]
The Results (Hepatocyte proliferation; page not available) report that phenobarbital induced proliferation in rat liver microtissues but not in human liver microtissues, supporting a narrower human-hepatocyte comparison. The paper does not establish that the c-Myc–Foxm1 axis itself was absent in human hepatocytes, and the retrieved passages do not address humanized CAR transgenic mice. (LLM, conf=0.84)
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Proteomic analysis of hepatic effects of phenobarbital in mice with humanized liver
Sprenger, H., Rasinger, J. D., Hammer, H., Naboulsi, W., Zabinsky, E., Planatscher, H., Schwarz, M., Poetz, O., & Braeuning, A. (2022) · Archives of Toxicology · doi:10.1007/s00204-022-03338-7 · Cited by 12 · see citing works
Zhou, W., Chembazhi, U. V., Huang, J., Bangru, S., Dean, A. E., Asokakumar, A., Kalsotra, A., Rudnick, D. A., & Anakk, S. (2026) · Cellular and Molecular Gastroenterology and Hepatology · doi:10.1016/j.jcmgh.2026.101858
- Authors
- Plummer, S., Beaumont, B., Wallace, S., Ball, G., Wright, J., McInnes, L., Currie, R., Peffer, R., & Cowie, D.
- Affiliations
- University of Dundee (GB)
- Syngenta (United Kingdom) (GB)
- Corresponding author
- Plummer, Simon
- Venue
- Toxicology Reports
- Publisher
- Elsevier BV
- Published
- 2019
- Volume
- 6
- Pages
- 998-1005
- ISSN
- 2214-7500 (ISSN-L)
- PMC
- PMC6816142
- OpenAlex
- W2976437814
- Cited by
- 20
- References
- 2
- Citations by year
- 2026: 3 · 2025: 4 · 2024: 4 · 2023: 1 · 2022: 2 · 2021: 3 · 2020: 3
- FWCI
- 0.63
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Toxicology Reports (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Discovery Research Portal (University of Dundee) (publishedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Carcinogens and Genotoxicity Assessment
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Pharmacogenetics and Drug Metabolism
- Life Sciences › Pharmacology, Toxicology and Pharmaceutics › Pharmacology › Drug-Induced Hepatotoxicity and Protection
- Keywords
- constitutive androstane receptor, phenobarbital, hepatocyte proliferation, enzyme induction, CYP2B6, CYP3A7, UGT1A6, non-parenchymal cells, cross-species comparison, mechanism of action, hepatocellular adenoma, hepatocellular carcinoma, transcriptomics, proteomics
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