Gut-liver-on-a-chip Enables Mechanistic Study and Risk Assessment of Drug-Induced Liver Injury and Drug-Drug Interactions

Abstract

Current preclinical models face challenges in recapitulating organ-level interactions affecting drug safety and there has been little investigation into drug toxicity and related DILI. We presented a pump-less gut-liver-on-chip enabling integrated analysis of drug exposure-toxicity relationships and inter-organ pharmacological interactions. The platform incorporated a polarized intestinal barrier with a quadruple cell co-cultured liver spheroid. Through simulation and comparative evaluation of oral versus systemic drug administration, we demonstrated the intestinal barrier's critical role in modulating drug exposure, corresponding toxic responses and first-pass effects. Temporal profiling revealed progressive hepatic injury mechanisms involving mitochondrial dysfunction and apoptotic pathway activation. Pharmacological inhibition of cytochrome P450 attenuated victim-induced oxidative stress without affecting hepatic drug exposure, confirming enzyme-related bioactivation as the toxicity mechanism. Furthermore, transporter-mediated drug-drug interactions were functionally recapitulated, with perpetrator compounds altering substrate pharmacokinetics through competitive efflux inhibition and modified intestinal disposition. The platform's ability to monitor drug exposure-toxicity relationships and drug-drug interactions was validated using combinations of perpetrator and victim drugs. This integrated approach advances organ-on-chips’ applications by establishing causal relationships between drug exposure and toxicity, resolving temporal toxicity progression, and modeling drug-drug interactions, which are critical factors in predicting clinical hepatotoxicity and complex pharmacokinetic interactions.