The immune phenotype of a tumour is a key predictor of its response to immunotherapy^1,2,3,4. Patients who respond to checkpoint blockade generally present with immune-inflamed^5,6,7 tumours that are highly infiltrated by T cells. However, not all inflamed tumours respond to therapy, and even lower response rates occur among tumours that lack T cells (immune desert) or that spatially exclude T cells to the periphery of the tumour lesion (immune excluded)⁸. Despite the importance of these tumour immune phenotypes in patients, little is known about their development, heterogeneity or dynamics owing to the technical difficulty of tracking these features in situ. Here we introduce skin tumour array by microporation (STAMP)—a preclinical approach that combines high-throughput time-lapse imaging with next-generation sequencing of tumour arrays. Using STAMP, we followed the development of thousands of arrayed tumours in vivo to show that tumour immune phenotypes and outcomes vary between adjacent tumours and are controlled by local factors within the tumour microenvironment. Particularly, the recruitment of T cells by fibroblasts and monocytes into the tumour core was supportive of T cell cytotoxic activity and tumour rejection. Tumour immune phenotypes were dynamic over time and an early conversion to an immune-inflamed phenotype was predictive of spontaneous or therapy-induced tumour rejection. Thus, STAMP captures the dynamic relationships of the spatial, cellular and molecular components of tumour rejection and has the potential to translate therapeutic concepts into successful clinical strategies.

肿瘤的免疫表型是其对免疫治疗反应的关键预测因素^1,2,3,4。对检查点封锁有反应的患者通常会出现免疫发炎的^5、6、7肿瘤，这些肿瘤被 T 细胞高度浸润。然而，并非所有发炎的肿瘤都对治疗有反应，在缺乏 T 细胞（免疫沙漠）或在空间上将 T 细胞排除在肿瘤病灶周围（免疫排除）的肿瘤中，反应率甚至更低⁸。尽管这些肿瘤免疫表型对患者很重要，但由于原位追踪这些特征的技术困难，人们对它们的发展、异质性或动态知之甚少。在这里，我们介绍微孔化皮肤肿瘤阵列（STAMP）——一种将高通量延时成像与下一代肿瘤阵列测序相结合的临床前方法。使用 STAMP，我们跟踪了体内数千个阵列肿瘤的发育，结果表明肿瘤免疫表型和结果在相邻肿瘤之间存在差异，并受肿瘤微环境中的局部因素控制。特别是，成纤维细胞和单核细胞将 T 细胞募集到肿瘤核心，支持 T 细胞的细胞毒活性和肿瘤排斥。肿瘤免疫表型随着时间的推移呈动态变化，早期转化为免疫炎症表型可预测自发性或治疗诱导的肿瘤排斥。因此，STAMP 捕捉了肿瘤排斥的空间、细胞和分子成分的动态关系，并有可能将治疗概念转化为成功的临床策略。