Scientist II Thermo Fisher Scientific Westminster, Maryland, United States
Introduction: Patient-derived 3D cancer models effectively mimic genomic and phenotypic characteristics of human tumors. Multiple studies have demonstrated that these models mirror the treatment response of the patients they originate from, highlighting their potential in drug development and, potentially, in screening therapies for personalized medicine. Here, we demonstrate the compatibility of tumoroids expanded in OncoPro ™ Tumoroid Culture Medium with compound screens using both OncoPro ™ and Human Plasma-Like Medium (HPLM). Interestingly, the potency of compounds across multiple drug classes varied depending on the chemical microenvironment (cell culture media) in which the assay was performed.
Materials and
Methods: Patient-derived tumoroids were cultured in OncoPro™ Tumroid Culture Medium before being dissociated and plated for drug screening assays. Colorectal cancer tumoroid lines from unique donors were seeded in either complete OncoPro™ medium or a screening medium in which HPLM was substituted for the OncoPro™ basal medium during formulation. Tumoroids were cultured in their respective medium in 96-well plates for three days prior to exposure to pharmacological compounds. The drug response assessment in these experiments utilized a plate reader-based assay to quantify ATP levels after 3 days of compound exposure, with subsequent calculation of IC50 values.
Results, Conclusions, and Discussions: Variations in the efficacy of compounds were observed depending on the basal medium used in the screening assay. Across multiple tumoroid lines, the microtubule destabilizer rigosertib had higher efficacy when screens were performed in medium formulated with the OncoPro™ basal compared to HPLM, while IC50 values were comparably lower when HPLM-based medium was used during testing of the antimetabolite azaserine. Dose response curves for clofarabine were comparable in each system. Literature suggests that these differences are due to the presence or absence of components in one basal medium versus the other. For example, uric acid present at physiologically-relevant levels in HPLM may interfere with microtubule destabilization caused by rigosertib in classical basal media (Rawat et al., 2023).
In conclusion, our findings emphasize the importance of selecting the most suitable basal medium, which can vary based on research aims and should be carefully considered during experimental design. This critical choice helps ensure robust and relevant outcomes, ultimately helping to advance our understanding of the translation between the response of patient-derived tumoroids and clinical relevance in the context of compound sensitivity screening.
Acknowledgements (Optional):
References: Rawat V, DeLear P, Prashanth P, Ozgurses ME, Tebeje A, Burns PA, Conger KO, Solís C, Hasnain Y, Novikova A, Endress JE, González-Sánchez P, Dong W, Stephanopoulos G, DeNicola GM, Harris IS, Sept D, Mason FM, Coloff JL. Drug screening in human physiologic medium identifies uric acid as an inhibitor of rigosertib efficacy. bioRxiv [Preprint]. 2023 Jul 28:2023.07.26.550731. doi: 10.1101/2023.07.26.550731. PMID: 37546939; PMCID: PMC10402161.
