Anstett Victor, Heinzelmann Elisa, Piraino Francesco, Roch Aline, Chrisnandy Antonius, Norkin Maxim, Garnier Virginie, Homicsko Krisztian, Hoehnel-Ka Sylke, Brandenberg Nathalie
Doppl SA, 1015 Lausanne, Switzerland.
Department of Oncology, CHUV, 1066 Lausanne, Switzerland.
Cancers (Basel). 2025 Jan 17;17(2):291. doi: 10.3390/cancers17020291.
: Developing ex vivo models that replicate immune-tumor interactions with high fidelity is essential for advancing immunotherapy research, as traditional two-dimensional in vitro systems often lack the complexity required to fully represent these interactions. : In this study, we establish a comprehensive 3D redirect lysis (3D-RDL) assay using colorectal cancer spheroids and adult stem cell-derived, healthy human organoids to evaluate the efficacy and safety profile of , a bispecific antibody targeting carcinoembryonic antigens (CEAs) on cancer cells and CD3 on T cells. This model allows us to assess cytotoxic activity and immune responses, capturing variations in therapeutic response not observable in simpler systems. Our model integrates live imaging and cytotoxicity analyses to enable precise, real-time tracking of antibody effects on CEA-expressing tumor cells compared to healthy cells. Additionally, by standardizing effector-to-target cell ratios in each co-culture, we establish a reproducible workflow that enhances data accuracy and comparability across assays. Flow cytometry and Granzyme B release profiling further allow us to characterize immune cell activation, revealing distinct T cell activation markers and Granzyme B release patterns tied to treatment. : Our results show that effectively induces cell death in cancer spheroids with high CEA expression while being dose-dependent on target, off-tumor binding and killing on non-cancerous cells of healthy organoids with intermediate CEA levels. This highlights our model's potential to predict clinical immunotherapy outcomes, capturing complex responses like immune activation, therapeutic selectivity, and potential resistance mechanisms. : These findings underscore the utility of our model as a reliable, physiologically relevant tool for screening new immunotherapies and advancing our understanding of tumor-immune dynamics.
开发能够高度逼真地复制免疫-肿瘤相互作用的体外模型对于推进免疫治疗研究至关重要，因为传统的二维体外系统往往缺乏充分展现这些相互作用所需的复杂性。在本研究中，我们使用结直肠癌球体和成人干细胞衍生的健康人类类器官建立了一种全面的3D重定向裂解（3D-RDL）检测方法，以评估一种双特异性抗体的疗效和安全性概况，该抗体靶向癌细胞上的癌胚抗原（CEA）和T细胞上的CD3。这个模型使我们能够评估细胞毒性活性和免疫反应，捕捉在更简单系统中无法观察到的治疗反应差异。我们的模型整合了实时成像和细胞毒性分析，以便与健康细胞相比，能够精确、实时地追踪抗体对表达CEA的肿瘤细胞的作用。此外，通过在每个共培养物中标准化效应细胞与靶细胞的比例，我们建立了一个可重复的工作流程，提高了数据准确性和不同检测之间的可比性。流式细胞术和颗粒酶B释放分析进一步使我们能够表征免疫细胞激活，揭示与治疗相关的独特T细胞激活标志物和颗粒酶B释放模式。我们的结果表明，该抗体在高表达CEA的癌球体中有效诱导细胞死亡，同时对具有中等CEA水平的健康类器官的非癌细胞的脱靶结合和杀伤呈剂量依赖性。这突出了我们的模型预测临床免疫治疗结果的潜力，捕捉免疫激活、治疗选择性和潜在耐药机制等复杂反应。这些发现强调了我们的模型作为一种可靠的、生理相关的工具在筛选新的免疫疗法和增进我们对肿瘤-免疫动力学理解方面的实用性。
