RNA-seq analysis of gene regulations by the combination treatment of crizotinib plus cisplatin in established MCA205 tumors. RNA-seq analysis of gene regulations by the combination treatment of crizotinib plus cisplatin in established MCA205 tumors

Immunogenic cell death (ICD) converts dying cancer cells into a therapeutic vaccine and stimulates antitumor immune responses. Here we unravel the results of an unbiased screen identifying high-dose (10 µM) crizotinib as an ICD-inducing tyrosine kinase inhibitor that has exceptional antineoplastic activity when combined with non-ICD inducing chemotherapeutics like cisplatin. The combination of cisplatin and high-dose crizotinib induces ICD in non-small cell lung carcinoma (NSCLC) cells and effectively controls the growth of distinct (transplantable, carcinogen- or oncogene induced) orthotopic NSCLC models. These anticancer effects are linked to increased T lymphocyte infiltration and are abolished by T cell depletion or interferon-g neutralization. Crizotinib plus cisplatin leads to an increase in the expression of PD-1 and PD-L1 in tumors, coupled to a strong sensitization of NSCLC to immunotherapy with PD-1 antibodies. Hence, a sequential combination treatment consisting in conventional chemotherapy together with crizotinib, followed by immune checkpoint blockade may be active against NSCLC. Overall design: Two groups: 4 replicats of control samples; 4 replicats of cisplatin plus crizotinib treated samples

免疫原性细胞死亡（Immunogenic cell death, ICD）可将濒死癌细胞转化为治疗性疫苗，并激活抗肿瘤免疫应答。本研究解析了一项无偏筛选的结果：高剂量（10 μM）克唑替尼（crizotinib）是一种可诱导免疫原性细胞死亡的酪氨酸激酶抑制剂（tyrosine kinase inhibitor），当其与顺铂（cisplatin）等非免疫原性细胞死亡诱导型化疗药物联用时，可展现优异的抗肿瘤活性。顺铂与高剂量克唑替尼的联合方案可在非小细胞肺癌（non-small cell lung carcinoma, NSCLC）细胞中诱导免疫原性细胞死亡，并有效抑制不同类型（可移植性、致癌物诱导或癌基因诱导）的原位非小细胞肺癌模型的肿瘤生长。此类抗癌效应与肿瘤内T淋巴细胞浸润增加密切相关，且可被T细胞耗竭或干扰素-γ中和作用完全阻断。克唑替尼联合顺铂可上调肿瘤组织中程序性死亡受体1（PD-1）与程序性死亡受体-配体1（PD-L1）的表达，同时可显著增强非小细胞肺癌对PD-1抗体免疫治疗的敏感性。因此，先采用常规化疗联合克唑替尼、后续予以免疫检查点阻断的序贯联合治疗策略，或许可对非小细胞肺癌产生临床疗效。整体实验设计：设置两组样本，对照组样本4个生物学重复，顺铂联合克唑替尼处理组样本4个生物学重复