Molecular Targeted Therapies Elicit Concurrent Apoptotic and GSDME-Dependent Pyroptotic Tumor Cell Death
Molecular Targeted Therapies Elicit Concurrent Apoptotic and GSDME-Dependent Pyroptotic Tumor Cell Death
复制标题
分子靶向治疗引发并发凋亡和 GSDME 依赖性焦亡肿瘤细胞死亡
DOI：
10.1158/1078-0432.ccr-18-1478
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发表时间：
2018-12-01
影响因子：
11.5
通讯作者：
Zhuang, Guanglei
中科院分区：
文献类型：
--
作者：
Lu, Haijiao;Zhang, Shengzhe;Zhuang, Guanglei
目的：癌基因抑制后诱导的死亡信号是分子靶向治疗人类癌症临床疗效的基础，而完整细胞凋亡机制的缺陷常常导致治疗失败。尽管具有潜在的重要性，但药物干预引发的其他形式的调节细胞死亡尚未得到系统的表征。实验设计：通过免疫印迹分析、相差成像、扫描电子显微镜和流式细胞术评估焦亡细胞死亡。使用 IHC 分析肺癌患者的肿瘤组织。在细胞系和异种移植模型中研究了细胞焦亡对药物反应的功能影响。结果：我们发现，专门针对 KRAS、EGFR 或 ALK 驱动的肺癌的多种小分子抑制剂除了同时引发细胞凋亡外，还一致引发强烈的焦亡细胞死亡。药物治疗后，线粒体内在凋亡途径被激活，动员的 caspase-3 蛋白酶裂解并激活 Gasdermin E（GSDME，由 DFNA5 编码），从而透化细胞质膜并执行细胞裂解性焦亡。 GSDME 在各种肺癌细胞系和临床标本中普遍表达，包括 KRAS 突变、EGFR 改变和 ALK 重排腺癌。结果，细胞凋亡和焦亡在肺癌细胞中广泛存在并相互作用，屈服于基因型匹配的治疗方案。我们进一步证明，焦亡细胞死亡在一定程度上促进了一部分癌症模型的药物反应。结论：这些结果表明，GSDME 依赖性细胞焦亡是分子靶向药物消除致癌基因成瘾性肿瘤细胞的一种先前未被认识的作用机制，这可能对抗癌治疗的临床开发和最佳应用具有重要意义。
Purpose: The induced death signals following oncogene inhibition underlie clinical efficacy of molecular targeted therapies against human cancer, and defects of intact cell apoptosis machinery often lead to therapeutic failure. Despite potential importance, other forms of regulated cell death triggered by pharmacologic intervention have not been systematically characterized. Experimental Design: Pyroptotic cell death was assessed by immunoblot analysis, phase-contrast imaging, scanning electron microscopy, and flow cytometry. Tumor tissues of patients with lung cancer were analyzed using IHC. Functional impact of pyroptosis on drug response was investigated in cell lines and xenograft models. Results: We showed that diverse small-molecule inhibitors specifically targeting KRAS-, EGFR-, or ALK-driven lung cancer uniformly elicited robust pyroptotic cell death, in addition to simultaneously invoking cellular apoptosis. Upon drug treatment, the mitochondrial intrinsic apoptotic pathway was engaged and the mobilized caspase-3 protease cleaved and activated gasdermin E (GSDME, encoded by DFNA5), which permeabilized cytoplasmic membrane and executed cell-lytic pyroptosis. GSDME displayed ubiquitous expression in various lung cancer cell lines and clinical specimens, including KRAS-mutant, EGFR-altered, and ALK-rearranged adenocarcinomas. As a result, cooccurrence and interplay of apoptosis and pyroptosis were widespread in lung cancer cells, succumbing to genotype-matched regimens. We further demonstrated that pyroptotic cell death partially contributed to the drug response in a subset of cancer models. Conclusions: These results pinpoint GSDME-dependent pyroptosis as a previously unrecognized mechanism of action for molecular targeted agents to eradicate oncogene-addicted neoplastic cells, which may have important implications for the clinical development and optimal application of anticancer therapeutics.