Hepatocellular carcinoma organoids enable drug discovery and combination therapy across heterogeneous tumors.

Hepatocellular carcinoma (HCC) remains a leading cause of cancer mortality, with limited therapeutic options and poor clinical outcomes. Tumor heterogeneity is a defining feature of HCC and a major barrier to effective treatment. To address this, we established a biobank of patient-derived HCC organoids generated from diagnostic biopsies, capturing the diversity of clinical features across disease stages. Using a high-throughput phenotypic screening platform, we tested more than 1,600 compounds and identified multiple agents with strong antitumor activity, including candidates suitable for drug repurposing. To further improve therapeutic responses, we systematically evaluated rationally designed doublet and triplet combinations anchored on regorafenib. Several combinations showed enhanced efficacy across HCCOs while maintaining selectivity over non-tumoral cells. In vivo validation of a selected triplet confirmed improved tumor control and tolerability. Together, these findings establish patient-derived organoids as a scalable platform for functional drug discovery and support combination strategies improving response across heterogeneous HCC.