Predicting the toxicity of cancer immunotherapies preclinically is challenging because models of tumours and healthy organs do not typically fully recapitulate the expression of relevant human antigens. Here we show that patient-derived intestinal organoids and tumouroids supplemented with immune cells can be used to study the on-target off-tumour toxicities of T-cell-engaging bispecific antibodies (TCBs), and to capture clinical toxicities not predicted by conventional tissue-based models as well as inter-patient variabilities in TCB responses. We analysed the mechanisms of T-cell-mediated damage of neoplastic and donor-matched healthy epithelia at a single-cell resolution using multiplexed immunofluorescence. We found that TCBs that target the epithelial cell-adhesion molecule led to apoptosis in healthy organoids in accordance with clinical observations, and that apoptosis is associated with T-cell activation, cytokine release and intra-epithelial T-cell infiltration. Conversely, tumour organoids were more resistant to damage, probably owing to a reduced efficiency of T-cell infiltration within the epithelium. Patient-derived intestinal organoids can aid the study of immune–epithelial interactions as well as the preclinical and clinical development of cancer immunotherapies.

临床前预测癌症免疫疗法的毒性具有挑战性，因为肿瘤和健康器官模型通常不能完全重现相关人类抗原的表达。在这里，我们表明，补充有免疫细胞的患者来源的肠道类器官和类肿瘤可用于研究 T 细胞接合双特异性抗体 (TCB) 的脱靶毒性，并捕获常规组织无法预测的临床毒性基于模型以及 TCB 反应的患者间差异。我们使用多重免疫荧光以单细胞分辨率分析了 T 细胞介导的肿瘤和供体匹配的健康上皮损伤的机制。根据临床观察，我们发现靶向上皮细胞粘附分子的 TCB 会导致健康类器官细胞凋亡，并且细胞凋亡与 T 细胞激活、细胞因子释放和上皮内 T 细胞浸润有关。相反，肿瘤类器官对损伤的抵抗力更强，这可能是由于上皮内 T 细胞浸润的效率降低。患者来源的肠道类器官可以帮助研究免疫-上皮相互作用以及癌症免疫疗法的临床前和临床开发。