Integration and validation of complementary ex vivo assays for functional precision oncology.

Functional precision oncology (FPO) enables individualized therapy selection using patient-derived tumor models, yet the concordance of distinct ex vivo testing strategies remains unclear. Here, we compare two orthogonal drug sensitivity platforms across molecularly characterized models spanning diverse pediatric (n = 13) and adult (n = 6) cancers. A rapid ATP-based assay quantifies viability within three days, whereas a long-term dynamic image-based platform captures microtumor dynamics over two weeks, incorporating pharmacokinetic features. Up to 50 drugs were profiled across both platforms, with additional combinations evaluated in the long-term assay; genomics-guided targeted drugs served as benchmarks. Both approaches robustly distinguished responders from non-responders and showed strong agreement in therapeutic prioritization (96.5% within 95% limits of agreement). The long-term dynamic platform achieved 81% sensitivity and 78% specificity, while resolving response depth and distinguishing cytostatic from cytotoxic effects. A representative sarcoma case highlights clinical relevance: long-term dynamic profiling predicted disease progression, whereas the short-term assay captured early treatment-associated viability effects. These findings establish cross-platform reproducibility in FPO and provide a systematic benchmarking of such approaches. Defining their complementary utility will be essential for integrating FPO strategies into clinical decision-making.