Next-Generation Sequencing of Pulmonary Sarcomatoid Carcinoma Reveals High Frequency of Actionable MET Gene Mutations
Next-Generation Sequencing of Pulmonary Sarcomatoid Carcinoma Reveals High Frequency of Actionable MET Gene Mutations
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DOI：
10.1200/jco.2015.62.0674
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发表时间：
2016-03-10
影响因子：
45.3
通讯作者：
Borczuk, Alain C.
中科院分区：
文献类型：
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作者：
Liu, Xuewen;Jia, Yuxia;Borczuk, Alain C.
目的进一步了解肺肉瘤样癌（pulmonary sarcomatoid carcinoma， PSC）的分子发病机制，为这种难治性疾病制定新的治疗策略。材料和方法对发现组（n = 10）进行全外显子组测序，对PSC独立验证组（n = 26）进行靶向MET突变筛选。通过逆转录酶聚合酶链反应和Western blotting验证MET外显子14的跳脱。在肺腺鳞细胞系H596 （MET外显子14被跳过，PIK3CA突变）和胃腺癌细胞系Hs746T （MET外显子14被跳过）中进行了功能研究，以验证MET外显子14跳过的致癌作用。我们对一名晚期PSC和MET外显子14跳脱患者对MET抑制剂治疗的反应进行了评估，以评估临床可转译性。结果除了确认已知癌症相关基因（TP53、KRAS、PIK3CA、MET、NOTCH、STK11和RB1）的突变外，还鉴定并验证了RASA1、CDH4、CDH7、LAMB4、SCAF1和LMTK2等其他基因的新突变。在36例患者中，有8例（22%）发现MET突变导致外显子14跳变；其中一个肿瘤也同时携带PIK3CA突变。短干扰RNA沉默MET和克唑替尼抑制MET对Hs746T和H596细胞的细胞活力有显著影响，下游AKT和丝裂原活化蛋白激酶活性降低。同时发生的PIK3CA突变需要添加第二种药物才能成功抑制途径并影响细胞活力。在一名携带MET外显子14跳变的晚期化疗难治性PSC患者中发现了对克唑替尼的显著反应。结论MET突变事件导致外显子14跳变是PSC中常见且潜在的靶向事件。(C) 2015年由美国临床肿瘤学会出版
PurposeTo further understand the molecular pathogenesis of pulmonary sarcomatoid carcinoma (PSC) and develop new therapeutic strategies in this treatment-refractory disease.Materials and MethodsWhole-exome sequencing in a discovery set (n = 10) as well as targeted MET mutation screening in an independent validation set (n = 26) of PSC were performed. Reverse transcriptase polymerase chain reaction and Western blotting were performed to validate MET exon 14 skipping. Functional studies for validation of the oncogenic roles of MET exon 14 skipping were conducted in lung adenosquamous cell line H596 (MET exon 14 skipped and PIK3CA mutated) and gastric adenocarcinoma cell line Hs746T (MET exon 14 skipped). Response to MET inhibitor therapy with crizotinib in a patient with advanced PSC and MET exon 14 skipping was evaluated to assess clinical translatability.ResultsIn addition to confirming mutations in known cancer-associated genes (TP53, KRAS, PIK3CA, MET, NOTCH, STK11, and RB1), several novel mutations in additional genes, including RASA1, CDH4, CDH7, LAMB4, SCAF1, and LMTK2, were identified and validated. MET mutations leading to exon 14 skipping were identified in eight (22%) of 36 patient cases; one of these tumors also harbored a concurrent PIK3CA mutation. Short interfering RNA silencing of MET and MET inhibition with crizotinib showed marked effects on cell viability and decrease in downstream AKT and mitogen-activated protein kinase activation in Hs746T and H596 cells. Concurrent PIK3CA mutation required addition of a second agent for successful pathway suppression and cell viability effect. Dramatic response to crizotinib was noted in a patient with advanced chemotherapy-refractory PSC carrying a MET exon 14 skipping mutation.ConclusionMutational events of MET leading to exon 14 skipping are frequent and potentially targetable events in PSC. (C) 2015 by American Society of Clinical Oncology