Silencing FLI or targeting CD13/ANPEP lead to dephosphorylation of EPHA2, a mediator of BRAF inhibitor resistance, and induce growth arrest or apoptosis in melanoma cells.
Silencing FLI or targeting CD13/ANPEP lead to dephosphorylation of EPHA2, a mediator of BRAF inhibitor resistance, and induce growth arrest or apoptosis in melanoma cells.
复制标题
DOI：
10.1038/cddis.2017.406
复制
发表时间：
2017-08-31
影响因子：
9
通讯作者：
Egyhazi Brage S
中科院分区：
文献类型：
--
作者：
Azimi A;Tuominen R;Costa Svedman F;Caramuta S;Pernemalm M;Frostvik Stolt M;Kanter L;Kharaziha P;Lehtiö J;Hertzman Johansson C;Höiom V;Hansson J;Egyhazi Brage S
大多数BRAF突变转移性黑色素瘤患者对BRAF抑制剂（BRAFi）治疗有反应，但由于获得性耐药，复发很常见。为了阐明BRAFi抗性机制，我们对敏感的亲本A375 BRAF V600 E突变的人黑素瘤细胞系和具有诱导的BRAFi抗性的子细胞系进行了基于基因表达和质谱的蛋白质组分析。在BRAFi耐药子细胞系中观察到两种新的耐药候选物氨基肽酶-N（CD 13/ANPEP）和ETS转录因子FLI 1的表达增加。此外，还发现了先前报告的耐药介质受体酪氨酸激酶受体A2（EPHA 2）和肝细胞生长因子受体MET水平升高。在BRAFi之前和疾病进展之后获得的来自黑素瘤患者的匹配肿瘤样品中评估这些蛋白质的表达。MET在所有进展样本中过表达，而其他候选者的表达在个体患者之间变化。通过阻断抗体靶向CD 13/ANPEP在亲本A375-和BRAFi-抗性子细胞以及具有内在BRAFi抗性的黑素瘤细胞中诱导凋亡，并导致S897上EPHA 2的去磷酸化，先前证明引起迁移能力的抑制。据报道，AKT和RSK均诱导EPHA 2 S897磷酸化，在抑制CD 13/ANPEP后也去磷酸化。FLI 1沉默还导致EPHA 2 S897磷酸化和总MET蛋白表达的降低。此外，FLI 1的沉默使抗性细胞对BRAFi敏感。此外，我们表明BRAFi与多激酶抑制剂达沙替尼组合可以消除BRAFi抗性并降低EPHA 2 S897磷酸化和总FLI 1蛋白表达。这是第一份报告提出CD 13/ANPEP和FLI 1作为BRAF抑制的重要介质，具有作为BRAF难治性黑色素瘤药物靶点的潜力。
A majority of patients with BRAF-mutated metastatic melanoma respond to therapy with BRAF inhibitors (BRAFi), but relapses are common owing to acquired resistance. To unravel BRAFi resistance mechanisms we have performed gene expression and mass spectrometry based proteome profiling of the sensitive parental A375 BRAF V600E-mutated human melanoma cell line and of daughter cell lines with induced BRAFi resistance. Increased expression of two novel resistance candidates, aminopeptidase-N (CD13/ANPEP) and ETS transcription factor FLI1 was observed in the BRAFi-resistant daughter cell lines. In addition, increased levels of the previously reported resistance mediators, receptor tyrosine kinase ephrine receptor A2 (EPHA2) and the hepatocyte growth factor receptor MET were also identified. The expression of these proteins was assessed in matched tumor samples from melanoma patients obtained before BRAFi and after disease progression. MET was overexpressed in all progression samples while the expression of the other candidates varied between the individual patients. Targeting CD13/ANPEP by a blocking antibody induced apoptosis in both parental A375- and BRAFi-resistant daughter cells as well as in melanoma cells with intrinsic BRAFi resistance and led to dephosphorylation of EPHA2 on S897, previously demonstrated to cause inhibition of the migratory capacity. AKT and RSK, both reported to induce EPHA2 S897 phosphorylation, were also dephosphorylated after inhibition of CD13/ANPEP. FLI1 silencing also caused decreases in EPHA2 S897 phosphorylation and in total MET protein expression. In addition, silencing of FLI1 sensitized the resistant cells to BRAFi. Furthermore, we show that BRAFi in combination with the multi kinase inhibitor dasatinib can abrogate BRAFi resistance and decrease both EPHA2 S897 phosphorylation and total FLI1 protein expression. This is the first report presenting CD13/ANPEP and FLI1 as important mediators of resistance to BRAF inhibition with potential as drug targets in BRAFi refractory melanoma.
登录
查看更多内容
Epha2是vemurafenib耐药性和黑色素瘤中新型治疗靶标的介体。
影响因子：
28.2
作者：
Miao B;Ji Z;Tan L;Taylor M;Zhang J;Choi HG;Frederick DT;Kumar R;Wargo JA;Flaherty KT;Gray NS;Tsao H
通讯作者：
Tsao H
不依赖配体的EPHA2信号传导驱动靶向治疗介导的转移性黑色素瘤表型的采用。
影响因子：
28.2
作者：
Paraiso KH;Das Thakur M;Fang B;Koomen JM;Fedorenko IV;John JK;Tsao H;Flaherty KT;Sondak VK;Messina JL;Pasquale EB;Villagra A;Rao UN;Kirkwood JM;Meier F;Sloot S;Gibney GT;Stuart D;Tawbi H;Smalley KS
通讯作者：
Smalley KS
影响因子：
28.2
作者：
Held MA;Langdon CG;Platt JT;Graham-Steed T;Liu Z;Chakraborty A;Bacchiocchi A;Koo A;Haskins JW;Bosenberg MW;Stern DF
通讯作者：
Stern DF
影响因子：
11.2
作者：
Paraiso, Kim H. T.;Xiang, Yun;Smalley, Keiran S. M.
通讯作者：
Smalley, Keiran S. M.
影响因子：
2
作者：
Park, Sung-Won;Do, Hyun-Jin;Kim, Jae-Hwan
通讯作者：
Kim, Jae-Hwan