Urinary Tract Tumor Organoids Reveal Eminent Differences in Drug Sensitivities When Compared to 2-Dimensional Culture Systems

Abstract

Generation of organoids from urinary tract tumor samples was pioneered a few years ago. We generated organoids from two upper tract urothelial carcinomas and from one bladder cancer sample, and confirmed the expression of cytokeratins as urothelial antigens, vimentin as a mesenchymal marker, and fibroblast growth factor receptor 3 by immunohistochemistry. We investigated the dose response curves of two novel components, venetoclax versus S63845, in comparison to the clinical standard cisplatin in organoids in comparison to the corresponding two-dimensional cultures. Normal urothelial cells and tumor lines RT4 and HT1197 served as controls. We report that upper tract urothelial carcinoma cells and bladder cancer cells in two-dimensional cultures yielded clearly different sensitivities towards venetoclax, S63845, and cisplatin. Two-dimensional cultures were more sensitive at low drug concentrations, while organoids yielded higher drug efficacies at higher doses. In some two-dimensional cell viability experiments, colorimetric assays yielded different IC50 toxicity levels when compared to chemiluminescence assays. Organoids exhibited distinct sensitivities towards cisplatin and to a somewhat lesser extent towards venetoclax or S63845, respectively, and significantly different sensitivities towards the three drugs investigated when compared to the corresponding two-dimensional cultures. We conclude that organoids maintained inter-individual sensitivities towards venetoclax, S63845, and cisplatin. The preclinical models and test systems employed may bias the results of cytotoxicity studies.

Effects of Scaffolds on Urine- and Urothelial Carcinoma Tissue-Derived Organoids from Bladder Cancer Patients

“…Information on the patients and stages of the disease is summarized in Table 1 . The tissue was minced by blade and scissors followed by enzymatic degradation as described recently [ 17 , 18 ]. Debris was removed by passing the extract through a cell strainer with 70 µm mesh size and cells were washed by centrifugation (150 g, 7 min, 20 °C).…”

3D Tumor Models in Urology

“…Wei and colleagues found significant differences in the sensitivities of BCa-cells and PDOs to cisplatin, venetoclax (a new Bcl-2 inhibitor), and S63845 (a novel MCL1 inhibitor). They conclude that PDOs more likely reflect the in vivo situation of BCa, affecting the reliability of preclinical test systems [ 81 ]. Yu et al have already gone a step further in their studies and established PDOs from luminal and basal resection specimens of a muscle-invasive BCa.…”

Advanced Bladder Cancer: Changing the Treatment Landscape

“…In the first report on BC PDOs, they often retained parental tumor heterogeneity [ 53 ]. It has been demonstrated that urinary tract PDOs maintained inter-individual sensitivity towards targeting and cytotoxic agents [ 54 ], which is not always true for 2D primary culture.…”