Hanne Verswyvel

Chapter 3 │ Page 84 Figure 3: Synergy calculation for NTP-CDDP combinations. Heatmap shows synergy scores of viability reduction for the di erent NTP-CDDP combinations 24 hours and 48 hours post-treatment for the 3 HNSCC cell lines. Bliss synergy analysis demonstrated additivity for scores between -10 and +10, and synergy for scores ≥ 10. Analysis performed with the online tool ‘Synergy Finder’. Taken together, our results demonstrated a significant reduction in cell viability at both 24 and 48 hours, with strong additive to synergistic synergy scores. These findings highlight the favorable indications for a NTP-CDDP combination. 3.2. NTP-CDDP Treatment E icacy Is Validated in Patient-Derived Organoids for HNSCC To further support our promising data in the HNSCC spheroids, our goal was to validate the e ects of our novel NTP-CDDP treatment on tumor kinetics in patientderived HNSCC organoids (HNSCC-PDO) (Figure 4). This advanced system would allow us to assess whether the observed tumor inhibition patterns and responses seen in 3D spheroids were consistent in a far more advanced, physiologically relevant context. We first evaluated treatment e icacy of both NTP and CDDP separately in two distinct human HNSCC-PDOs, i.e., HNSCC_001T and HNSCC_007T. Four-day-old PDOs were treated with di erent regimes of NTP or a concentration of the 7-points titration of CDDP, and followed up for 72 hours with live-cell imaging. We observed a dose-dependent decrease in organoid viability for both entities. Furthermore, we could appreciate the heterogeneity captured in the HNSCC-PDO model, as the

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