Chapter 3 │ Page 71 (HNSCC-PDOs), highlighting the potential of this combination to improve treatment outcomes. We analyzed the timely expression of key DAMPs, cytokines, and chemokines, providing insights into the immune-stimulatory properties of NTP-CDDP therapy. Finally, we validated bona fide ICD induction and immune engagement with the gold-standard vaccination assay in a syngeneic mouse model for HNSCC, confirming the ability of this combination to elicit a robust anti-tumor immune response in a complete biological system. Altogether, our findings underscore the potential of NTP to enhance the immunogenic properties of CDDP, which remains the classical backbone of many HNSCC treatment courses. This data provides a strong foundation for the rational design of NTP-based combination strategies, aiming to improve therapeutic e icacy and patient response without increasing treatment-related burden. Ultimately, these insights could accelerate the clinical translation of NTP, o ering a novel, immune-stimulatory approach to enhance conventional HNSCC therapies. 2. MATERIAL AND METHODS 2.1. Cell Culture Three human and one mouse HNSCC cell line, Cal27, SCC22B and SCC61, and SCC7, were used in this study. The Cal27 cell line was purchased from American Type Culture Collection (ATCC, Rockville, MD, USA), and the SCC22B and SCC61 cell line were kindly provided by Prof. Dr. Olivier De Wever (Laboratory of Experimental Cancer Research, Ghent University Hospital, Ghent, Belgium) and Prof. Dr. Sandra Nuyts (University Hospital Leuven, Leuven, Belgium), respectively. The mouse HNSCC SCC7 cells were kindly provided by Prof. Dr. Reinhard Zeidler (Helmholtz Munich, Germany). Cell lines were commercially available, and no ethical approval was required.
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