Center for Medicine Research, Eberhard Karls University, 72072 Tuebingen, Germany.
Department of Urology, University Hospital, 72076 Tuebingen, Germany.
Int J Mol Sci. 2022 Jun 4;23(11):6305. doi: 10.3390/ijms23116305.
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 IC 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.
几年前，人们开创性地从尿路上皮肿瘤样本中生成类器官。我们从两个上尿路尿路上皮癌和一个膀胱癌样本中生成了类器官，并通过免疫组织化学证实了细胞角蛋白作为尿路上皮抗原、波形蛋白作为间充质标志物和成纤维细胞生长因子受体 3 的表达。我们研究了两种新型成分 venetoclax 和 S63845 与临床标准顺铂在类器官中的剂量反应曲线，与相应的二维培养物进行比较。正常尿路上皮细胞和肿瘤细胞系 RT4 和 HT1197 作为对照。我们报告称，在上尿路尿路上皮癌细胞和膀胱癌细胞的二维培养物中，venetoclax、S63845 和顺铂对其的敏感性明显不同。二维培养物在低药物浓度下更敏感，而类器官在更高剂量下产生更高的药物疗效。在一些二维细胞活力实验中，与化学发光测定法相比，比色法得出的 IC 毒性水平不同。类器官对顺铂表现出明显的敏感性，对 venetoclax 或 S63845 的敏感性略低，与相应的二维培养物相比，对三种研究药物的敏感性明显不同。我们得出结论，类器官保持了个体间对 venetoclax、S63845 和顺铂的敏感性。所使用的临床前模型和测试系统可能会影响细胞毒性研究的结果。
