Novel role of Engrailed 1 as a prosurvival transcription factor in basal-like breast cancer and engineering of interference peptides block its oncogenic function.
Novel role of Engrailed 1 as a prosurvival transcription factor in basal-like breast cancer and engineering of interference peptides block its oncogenic function.
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DOI：
10.1038/onc.2013.422
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发表时间：
2014-09-25
期刊：
影响因子：
8
通讯作者：
Blancafort P
中科院分区：
文献类型：
--
作者：
Beltran AS;Graves LM;Blancafort P
基底细胞样乳腺肿瘤是一种侵袭性肿瘤，具有高度的增殖和转移。化疗是目前唯一的治疗选择;然而，耐药性经常发生，导致复发和患者死亡。一些极具侵袭性的癌症也与缺氧、炎症和高白细胞浸润有关。在此，我们发现神经特异性转录因子Engrailed 1（EN 1）仅在这些肿瘤中过表达。短发夹RNA（shRNA）介导的EN 1敲低引发了有效的和选择性的细胞死亡。相反，正常细胞中EN 1的异位过表达激活了存活途径，并赋予了对化疗药物的抗性。外源性表达的EN 1 cDNA重新编程的乳腺上皮细胞向一个长寿的，神经样表型显示多巴胺能标记。基因表达微阵列表明，EN 1 cDNA改变了大量炎症分子的转录，特别是趋化因子和趋化因子受体，它们可以介导促生存途径。为了阻断EN 1功能，我们设计了合成干扰肽（iPeps），其包含EN 1特异性序列和N末端细胞穿透肽/核定位序列，所述序列介导EN 1功能所必需的基本蛋白质-蛋白质相互作用。这些EN 1-iPeps在过表达EN 1的肿瘤细胞中迅速介导强烈的凋亡反应，对正常或非EN 1表达细胞没有毒性。将EN 1-iPeps递送到基底样癌细胞中显著降低了常规用于治疗乳腺癌的化疗药物的50%抑制浓度（IC 50）。最后，基质辅助激光解吸/电离飞行时间质谱和免疫沉淀分析表明，EN 1-iPeps捕获的目标参与转录和转录后调控。重要的是，EN 1-iPeps结合谷氨酰-脯氨酰tRNA合成酶（EPRS）靶标，该靶标与炎症蛋白的转录特异性翻译控制和氨基酸应激途径的激活相关。这项工作揭示了EN 1作为与基底样乳腺癌中促生存相关的内在炎症通路的激活剂。我们进一步建立在这些结果的基础上，并描述了靶向EN 1（EN 1-iPeps）的iPeps工程作为一种新的和选择性的治疗策略，以打击这些致命形式的乳腺癌。
Basal-like breast tumors are aggressive cancers associated with high proliferation and metastasis. Chemotherapy is currently the only treatment option; however, resistance often occurs resulting in recurrence and patient death. Some extremely aggressive cancers are also associated with hypoxia, inflammation and high leukocyte infiltration. Herein, we discovered that the neural-specific transcription factor, Engrailed 1 (EN1), is exclusively overexpressed in these tumors. Short hairpin RNA (shRNA)-mediated knockdown of EN1 triggered potent and selective cell death. In contrast, ectopic overexpression of EN1 in normal cells activated survival pathways and conferred resistance to chemotherapeutic agents. Exogenous expression of EN1 cDNA reprogrammed the breast epithelial cells toward a long-lived, neural-like phenotype displaying dopaminergic markers. Gene expression microarrays demonstrated that the EN1 cDNA altered transcription of a high number of inflammatory molecules, notably chemokines and chemokine receptors, which could mediate prosurvival pathways. To block EN1 function, we engineered synthetic interference peptides (iPeps) comprising the EN1-specific sequences that mediate essential protein-protein interactions necessary for EN1 function and an N-terminal cell-penetrating peptide/nuclear localization sequence. These EN1-iPeps rapidly mediated a strong apoptotic response in tumor cells overexpressing EN1, with no toxicity to normal or non EN1-expressing cells. Delivery of EN1-iPeps into basal-like cancer cells significantly decreased the fifty percent inhibitory concentrations (IC50) of chemotherapeutic drugs routinely used to treat breast cancer. Lastly, matrix-assisted laser desorption/ionization-time of flight mass spectrometry and immunoprecipitation assays demonstrated that EN1-iPeps captured targets involved in transcriptional and post-transcriptional regulation. Importantly, the EN1-iPeps bound the glutamyl-prolyl tRNA synthetase (EPRS) target, which has been associated with the transcript-specific translational control of inflammatory proteins and activation of amino-acid stress pathways. This work unveils EN1 as an activator of intrinsic inflammatory pathways associated with prosurvival in basal-like breast cancer. We further build upon these results and describe the engineering of iPeps targeting EN1 (EN1-iPeps) as a novel and selective therapeutic strategy to combat these lethal forms of breast cancer.
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DOI：
10.1186/bcr554
发表时间：
2003
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Ben-Baruch A
通讯作者：
Ben-Baruch A
影响因子：
14.8
作者：
Keller, Tracy L.;Zocco, Davide;Sundrud, Mark S.;Hendrick, Margaret;Edenius, Maja;Yum, Jinah;Kim, Yeon-Jin;Lee, Hak-Kyo;Cortese, Joseph F.;Wirth, Dyann F.;Dignam, John David;Rao, Anjana;Yeo, Chang-Yeol;Mazitschek, Ralph;Whitman, Malcolm
通讯作者：
Whitman, Malcolm
影响因子：
2.7
作者：
Fan C;Prat A;Parker JS;Liu Y;Carey LA;Troester MA;Perou CM
通讯作者：
Perou CM
DOI：
10.1186/bcr3019
发表时间：
2011-09-27
期刊：
Breast cancer research : BCR
影响因子：
--
作者：
Beltran AS;Rivenbark AG;Richardson BT;Yuan X;Quian H;Hunt JP;Zimmerman E;Graves LM;Blancafort P
通讯作者：
Blancafort P
影响因子：
5.3
作者：
Arif, Abul;Chatterjee, Piyali;Fox, Paul L.
通讯作者：
Fox, Paul L.