肿瘤细胞 (TC) 上的 PD-L1 表达被用作肺癌的免疫治疗生物标志物，但经常观察到肿瘤内的异质表达。我们使用数字空间分析，根据肿瘤 PD-L1 表达和免疫微环境的状态，对空间匹配区域的 TC 和免疫细胞 (IC) 进行蛋白质组和全转录组分析。我们的研究结果通过免疫组织化学、癌症基因组图谱和免疫治疗队列进行了验证。 TC 上 PD-L1 高表达区域的 IC 比 TC 上 PD-L1 低表达区域的 IC 显示更多的免疫抑制和衰竭特征。在免疫炎症 (IF) 区域内高表达 PD-L1 的 TC 显示出促炎症过程的上调，而在免疫缺陷 (ID) 区域内高表达 PD-L1 的 TC 显示出各种代谢过程的上调。利用 IF 和 ID 区域之间 TC 的差异表达基因，我们确定了一种新的肺癌预后基因特征。此外，CD8 细胞与 M2 巨噬细胞的高比例被发现可以预测免疫检查点抑制剂治疗后表达 PD-L1 的肺癌患者的良好结果。这项研究表明，TC 和 IC 在肿瘤微环境中具有独特的空间特征，这些特征与肿瘤 PD-L1 表达和 IC 浸润相关。

The expression of PD-L1 on tumor cells (TCs) is used as an immunotherapy biomarker in lung cancer, but heterogeneous intratumoral expression is often observed. Using a Digital Spatial Profiling, we performed proteomic and whole-transcriptomic analyses of TCs and immune cells (ICs) in spatially matched areas based on tumor PD-L1 expression and the status of the immune microenvironment. Our findings were validated using immunohistochemistry, The Cancer Genome Atlas, and immunotherapy cohorts. ICs in areas with high PD-L1 expression on TCs showed more features indicative of immunosuppression and exhaustion than ICs in areas with low PD-L1 expression on TCs. TCs highly expressing PD-L1 within immune-inflamed (IF) areas show up-regulation of pro-inflammatory processes, whereas TCs highly expressing PD-L1 within immune-deficient (ID) areas show up-regulation of various metabolic processes. Using differentially expressed genes of TCs between the IF and ID areas, we identified a novel prognostic gene signature for lung cancer. In addition, a high ratio of CD8+ cells to M2 macrophages was found to predict favorable outcomes in patients with PD-L1-expressing lung cancer after immune checkpoint inhibitor therapy. This study demonstrates that TCs and ICs have distinct spatial features within the tumor microenvironment that are related to tumor PD-L1 expression and IC infiltration.