免疫逃避的驱动因素尚不完全清楚，这限制了癌症免疫疗法的成功。在这里，我们应用单细胞空间和微扰转录组学来描述高级别浆液性输卵管卵巢癌的免疫逃避。为此，我们首先通过对 94 名患者的 130 个肿瘤中超过 250 万个原位细胞进行分析，绘制了高级浆液性输卵管卵巢癌的空间组织图。这揭示了一种反映肿瘤遗传学的恶性细胞状态，并可预测 T 细胞和自然杀伤细胞的浸润水平以及对免疫检查点封锁的反应。然后，我们进行了 Perturb-seq 筛选并鉴定了触发这种恶性细胞状态的遗传扰动（包括敲除 PTPN1 和 ACTR8）。最后，我们表明，正如预测的那样，这些扰动以及 PTPN1/PTPN2 抑制剂使卵巢癌细胞对 T 细胞和自然杀伤细胞的细胞毒性敏感。因此，本研究确定了通过将遗传变异、细胞状态调节器和空间生物学联系起来来研究和瞄准免疫逃避的方法。© 2024。作者。

The drivers of immune evasion are not entirely clear, limiting the success of cancer immunotherapies. Here we applied single-cell spatial and perturbational transcriptomics to delineate immune evasion in high-grade serous tubo-ovarian cancer. To this end, we first mapped the spatial organization of high-grade serous tubo-ovarian cancer by profiling more than 2.5 million cells in situ in 130 tumors from 94 patients. This revealed a malignant cell state that reflects tumor genetics and is predictive of T cell and natural killer cell infiltration levels and response to immune checkpoint blockade. We then performed Perturb-seq screens and identified genetic perturbations-including knockout of PTPN1 and ACTR8-that trigger this malignant cell state. Finally, we show that these perturbations, as well as a PTPN1/PTPN2 inhibitor, sensitize ovarian cancer cells to T cell and natural killer cell cytotoxicity, as predicted. This study thus identifies ways to study and target immune evasion by linking genetic variation, cell-state regulators and spatial biology.© 2024. The Author(s).