Domingues Ana Carolina M, Palin Claire, Sun Yi, Xie Hongyan, Woods Elliot C, Jenkins Russell W, Revach Or-Yam
Mass General Cancer Center, Krantz Family Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States; Instituto do Câncer do Estado de São Paulo, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil.
Mass General Cancer Center, Krantz Family Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States.
Methods Cell Biol. 2025;196:139-159. doi: 10.1016/bs.mcb.2024.11.003. Epub 2024 Dec 14.
Patient-derived tumor models are emerging as promising tools to explore patient-specific tumor behavior and to address a central gap in tumor immunology and immunotherapy drug development - the need for clinically relevant tumor models that recapitulate the complexity of the human tumor ecosystem, in which cancer cells are interacting with various immune and stromal elements. Patient-derived organotypic tumor spheroids (PDOTS), a biomimetic 3D-patient tumor avatar (3D-PTA), are comprised of cancer cells and autologous tumor-infiltrating immune and stromal cells that are grown within collagen hydrogels embedded in a 3D microfluidic culture device to model physiologic conditions and enable the study of tumor-immune dynamics. PDOTS and their murine counterparts (MDOTS, murine-derived organotypic tumor spheroids) are responsive to immune checkpoint blockade (ICB) and mirror in vivo response dynamics. We have also confirmed the utility of MDOTS/PDOTS in examining novel therapeutic strategies to overcome ICB resistance and testing the efficiency of T cell-based immunotherapies, demonstrating the utility of PDOTS profiling in examining the tumor-immune dynamics of immunotherapy response and resistance. Here, we provide a detailed protocol for processing, ex vivo culture, and analysis of patient-derived tumor organotypic tumor spheroids (PDOTS) in 3D microfluidic culture for immuno-oncology applications. The protocol can be readily adapted for ex vivo profiling of murine-derived organotypic tumor spheroids (MDOTS) and cancer cell line-derived monotypic tumor spheroids (MTS) for robust and iterative testing of immuno-oncology targets.
患者来源的肿瘤模型正成为探索患者特异性肿瘤行为以及填补肿瘤免疫学和免疫治疗药物开发核心空白的有前景的工具，即需要临床相关的肿瘤模型来重现人类肿瘤生态系统的复杂性，其中癌细胞与各种免疫和基质成分相互作用。患者来源的器官型肿瘤球体（PDOTS），一种仿生的三维患者肿瘤替身（3D-PTA），由癌细胞以及自体肿瘤浸润免疫细胞和基质细胞组成，这些细胞在嵌入三维微流控培养装置的胶原蛋白水凝胶中生长，以模拟生理条件并能够研究肿瘤免疫动力学。PDOTS及其小鼠对应物（MDOTS，小鼠来源的器官型肿瘤球体）对免疫检查点阻断（ICB）有反应，并反映体内反应动力学。我们还证实了MDOTS/PDOTS在研究克服ICB耐药性的新治疗策略以及测试基于T细胞的免疫疗法效率方面的实用性，证明了PDOTS分析在研究免疫治疗反应和耐药性的肿瘤免疫动力学方面的实用性。在此，我们提供了一份详细的方案，用于在三维微流控培养中处理、体外培养和分析患者来源的肿瘤器官型肿瘤球体（PDOTS），以用于免疫肿瘤学应用。该方案可轻松适用于小鼠来源的器官型肿瘤球体（MDOTS）和癌细胞系来源的单型肿瘤球体（MTS）的体外分析，以对免疫肿瘤学靶点进行稳健且迭代的测试。
