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Updated: Jan 16, 2026
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An Intestine/Liver Microphysiological System for Drug Pharmacokinetic and Toxicological Assessment
Published on: December 3, 2020
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Single-cell transcriptomics of acetaminophen-induced responses in human 2D and 3D liver microtissues
Brian Bwanya1, Marcha C T Verheijen1, Duncan Hauser1
1Department of Translational Genomics, GROW Research Institute for Oncology and Developmental Biology, Maastricht University, 6229 ER, Maastricht, The Netherlands.
Archives of Toxicology
|January 14, 2026
Summary
Three-dimensional (3D) liver spheroids better model drug-induced liver injury than 2D cultures. Hypoxia in 3D spheroids impacts acetaminophen metabolism, revealing crucial insights for pharmaceutical development.
Area of Science:
- Hepatotoxicity and Drug Metabolism
- In Vitro Liver Models
- Cellular and Molecular Toxicology
Background:
- Drug-induced liver injury (DILI) is a significant challenge in drug development and a primary cause of acute liver failure.
- Predictive and human-relevant in vitro models are essential for evaluating liver toxicity.
- Acetaminophen serves as a benchmark for studying hepatotoxicity mechanisms due to its well-defined dose-dependent effects.
Purpose of the Study:
- To compare cellular responses to acetaminophen exposure in 2D and 3D human liver cell culture models using single-cell RNA sequencing.
- To investigate the influence of oxygen availability and microenvironment on drug metabolism and toxicity.
- To assess the physiological relevance of 3D liver spheroids for mechanistic toxicity studies.
Main Methods:
- Primary human hepatocytes, Kupffer cells, and liver endothelial cells were cultured in 2D monolayers and 3D spheroids.
- Cultures were exposed to low and high acetaminophen concentrations for 24 hours.
- Single-cell RNA sequencing and Reactome pathway analysis were employed to characterize cellular responses.
Main Results:
- 3D spheroids demonstrated greater transcriptional diversity and enhanced metabolic/biosynthetic activity compared to 2D cultures.
- 3D cultures exhibited significant hypoxia-associated signaling, particularly in endothelial cells, due to restricted oxygen diffusion.
- Acetaminophen exposure in hypoxic hepatocytes led to increased CYP450 enzyme expression and decreased detoxification enzyme activity, indicating compromised Phase II metabolism.
Conclusions:
- 3D liver spheroids offer superior physiological relevance for studying DILI compared to 2D models.
- Oxygen tension dynamically interacts with acetaminophen metabolism, influencing cellular responses and toxicity.
- Spatial microenvironmental context, including oxygen gradients, is critical for accurate mechanistic insights into hepatotoxicity.
