Robotic Generation and Kinetic Screening of Patient-Derived Glioblastoma Organoids Reveal Pharmaceutical Dynamics

Glioblastoma (GBM) presents a significant challenge in oncology due to its high heterogeneity and poor response to conventional therapies. Current preclinical models are highly variable, creating disparate patient-derived organoids. We developed a Robot-Directed Organoid Deposition (RODEO) system that reproducibly generates complex patient-derived GBM organoids for high-throughput screening. RODEO-GBM organoids recapitulate the cellular heterogeneity, specific tumor niches, and complex vascularization of GBM. Integrating tissue-embedded microsensors in RODEO-GBM organoids allowed us to screen 126 FDA-approved oncology drugs, on 1,638 patient-derived organoids, generating 105 data points in 96 hrs. Our screen revealed distinct kinetic patterns of drug responses, identifying transient as well as slow-acting compounds often overlooked by conventional methods. Slow-acting drugs preferentially targeted stem-like SOX2+ tumor cells, thus preserving peripheral cell populations. Comparative analysis on RODEO-Liver organoids showed that slow-acting drugs exhibit lower liver toxicity supporting a higher therapeutic index. Coupling RODEO with sensor-based kinetic screening provides critical new insights into pharmaceutical dynamics, paving the way for the identification of more effective treatments for GBM and other cancers.