Abstract 3152: Development of iPSC-derived human liver organoids for preclinical drug testing and toxicology studies

Abstract Background and Purpose: Over 90% of drug candidates fail during clinical trials, often due to insufficient efficacy and unmanageable toxicity. Drug-induced liver injury (DILI) is frequently missed in preclinical testing due to a lack of robust and predictive liver models. We report the de novo development of iPSC-derived human liver organoids as scalable, reliable, and physiologically relevant preclinical toxicology models. Methods: Using an efficient and reproducible iPSC-differentiation protocol, we generated long term expandable, cryopreservable bipotential 3dGROTM Human iPSC-derived Liver Progenitor Organoids, which differentiate into mature liver organoids (MLOs) containing multiple liver cell types. Results: The iPSC-derived mature liver organoids display long-term stability, secrete albumin and urea, and express key biomarkers of mature hepatocytes (albumin, CYP3A4, HNF4a, PCK2) and cholangiocytes (Sox17, Sox9, CK7, MRP2). They also exhibit functional Phase I/II liver enzymes (CYP3A4, CYP2C9, CYP1A2, ALT, AST, GST) and active bile salt/drug transporters. Compared to primary hepatocytes, our 3dGROTM Human iPSC-derived Mature Liver Organoids demonstrate comparable functionality and outperform HepG2 cells and liver spheroids. In DILI assays, MLOs responded to drug treatment with elevated liver enzymes, confirming their potential for toxicology screening. Conclusion: These liver organoids represent a powerful tool for high-throughput drug screening, DMPK and toxicological studies, with future applications including disease modeling of hepatocellular carcinoma, MASLD, and NASH. Citation Format: Fong Cheng Pan, Mahi Rahman, David Austin, Stephan Krieg, Luisa Marie Pfeifer, Anthony Saporita, Philip Hewitt, Steven Johnston, Laura Braeuninger-Weimer, Willem Kools, Vi Chu. Development of iPSC-derived human liver organoids for preclinical drug testing and toxicology studies [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 3152.