MEK is a canonical effector of mutant KRAS;however,MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers.Here,we identified mitochondrial oxidative phosphorylation(OXPHOS)induction as a pr...MEK is a canonical effector of mutant KRAS;however,MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers.Here,we identified mitochondrial oxidative phosphorylation(OXPHOS)induction as a profound metabolic alteration to confer KRAS-mutant non-small cell lung cancer(NSCLC)resistance to the clinical MEK inhibitor trametinib.Metabolic flux analysis demonstrated that pyruvate metabolism and fatty acid oxidation were markedly enhanced and coordinately powered the OXPHOS system in resistant cells after trametinib treatment,satisfying their energy demand and protecting them from apoptosis.As molecular events in this process,the pyruvate dehydrogenase complex(PDHc)and carnitine palmitoyl transferase IA(CPTIA),two rate-limiting enzymes that control the metabolic flux of pyruvate and palmitic acid to mitochondrial respiration were activated through phosphorylation and transcriptional regulation.Importantly,the co-administration of trametinib and IACS-010759,a clinical mitochondrial complex I inhibitor that blocks OXPHOS,significantly impeded tumor growth and prolonged mouse survival.Overall,our findings reveal that MEK inhibitor therapy creates a metabolic vulnerability in the mitochondria and further develop an effective combinatorial strategy to circumvent MEK inhibitors resistance in KRAS-driven NSCLC.展开更多
目的 探究Src同源2结构域蛋白酪氨酸磷酸酶(Src homology region 2-containing protein tyrosine phosphatase 2,SHP2)抑制剂对KRAS突变肺癌细胞中炎症相关基因表达的调节作用,旨在为KRAS突变肺癌治疗的个体化和精准化提供依据。方法 使...目的 探究Src同源2结构域蛋白酪氨酸磷酸酶(Src homology region 2-containing protein tyrosine phosphatase 2,SHP2)抑制剂对KRAS突变肺癌细胞中炎症相关基因表达的调节作用,旨在为KRAS突变肺癌治疗的个体化和精准化提供依据。方法 使用SHP2抑制剂SHP099处理5株KRAS突变的人肺癌细胞系,western blot分析ERK1/2等信号通路蛋白活化水平。采用RNA测序分析药物或DMSO作用后配对样本基因表达情况,对差异表达基因进行基因本体(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析揭示功能通路富集情况。采用RT-qPCR验证差异基因相对表达水平。结果 SHP2抑制剂处理后,KRAS突变肺癌细胞中的p-SHP2和p-ERK1/2表达水平显著下调。转录组测序和差异基因分析显示,SHP2靶向抑制改变了多个信号通路基因的表达,特别是白细胞介素17(interleukin 17,IL-17)通路相关的CXCL1、CXCL2、CXCL8、MMP1、PTGS2等炎症相关基因表达受到显著下调。结论 SHP2靶向抑制可影响KRAS突变肺癌细胞的IL-17信号通路炎症相关基因表达情况,为进一步阐述靶向SHP2调节肺癌免疫微环境提供了初步证据。展开更多
基金sponsored by the National Natural Science Foundation of China(82122045,82073073,81874207,and 81872418)Innovative Research Team of High-level Local Universities in Shanghai(SHSMU-ZDCX20210802,China)+4 种基金MOE Key Laboratory of Biosystems Homeostasis&Protection(Zhejiang University,China)Science and Technology Commission of Shanghai Municipality(21S11902000,China)Jointed PI Program from Shanghai Changning Maternity and Infant Health Hospital(11300-412311-20033,China)ECNU Construction Fund of Innovation and Entrepreneurship Laboratory(44400-20201-532300/021,China)the ECNU multifunctional platform for innovation(011 and 004,China).
文摘MEK is a canonical effector of mutant KRAS;however,MEK inhibitors fail to yield satisfactory clinical outcomes in KRAS-mutant cancers.Here,we identified mitochondrial oxidative phosphorylation(OXPHOS)induction as a profound metabolic alteration to confer KRAS-mutant non-small cell lung cancer(NSCLC)resistance to the clinical MEK inhibitor trametinib.Metabolic flux analysis demonstrated that pyruvate metabolism and fatty acid oxidation were markedly enhanced and coordinately powered the OXPHOS system in resistant cells after trametinib treatment,satisfying their energy demand and protecting them from apoptosis.As molecular events in this process,the pyruvate dehydrogenase complex(PDHc)and carnitine palmitoyl transferase IA(CPTIA),two rate-limiting enzymes that control the metabolic flux of pyruvate and palmitic acid to mitochondrial respiration were activated through phosphorylation and transcriptional regulation.Importantly,the co-administration of trametinib and IACS-010759,a clinical mitochondrial complex I inhibitor that blocks OXPHOS,significantly impeded tumor growth and prolonged mouse survival.Overall,our findings reveal that MEK inhibitor therapy creates a metabolic vulnerability in the mitochondria and further develop an effective combinatorial strategy to circumvent MEK inhibitors resistance in KRAS-driven NSCLC.
文摘目的 探究Src同源2结构域蛋白酪氨酸磷酸酶(Src homology region 2-containing protein tyrosine phosphatase 2,SHP2)抑制剂对KRAS突变肺癌细胞中炎症相关基因表达的调节作用,旨在为KRAS突变肺癌治疗的个体化和精准化提供依据。方法 使用SHP2抑制剂SHP099处理5株KRAS突变的人肺癌细胞系,western blot分析ERK1/2等信号通路蛋白活化水平。采用RNA测序分析药物或DMSO作用后配对样本基因表达情况,对差异表达基因进行基因本体(Gene Ontology,GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)富集分析揭示功能通路富集情况。采用RT-qPCR验证差异基因相对表达水平。结果 SHP2抑制剂处理后,KRAS突变肺癌细胞中的p-SHP2和p-ERK1/2表达水平显著下调。转录组测序和差异基因分析显示,SHP2靶向抑制改变了多个信号通路基因的表达,特别是白细胞介素17(interleukin 17,IL-17)通路相关的CXCL1、CXCL2、CXCL8、MMP1、PTGS2等炎症相关基因表达受到显著下调。结论 SHP2靶向抑制可影响KRAS突变肺癌细胞的IL-17信号通路炎症相关基因表达情况,为进一步阐述靶向SHP2调节肺癌免疫微环境提供了初步证据。
