High throughput kinase inhibitor screens reveal TRB3 and MAPK-ERK/TGFβ pathways as fundamental Notch regulators in breast cancer
High throughput kinase inhibitor screens reveal TRB3 and MAPK-ERK/TGFβ pathways as fundamental Notch regulators in breast cancer
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DOI：
10.1073/pnas.1214014110
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发表时间：
2013-01-29
影响因子：
11.1
通讯作者：
Reedijk, Michael
中科院分区：
文献类型：
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作者：
Izrailit, Julia;Berman, Hal K.;Reedijk, Michael
Notch配体Jagged 1（JAG 1）的表达和Notch激活促进乳腺癌的不良预后。我们使用高通量筛选来鉴定在这种情况下负责Notch激活的元件。化学激酶抑制剂和激酶特异性小干扰RNA文库在乳腺癌细胞系中筛选，该细胞系经工程改造以报告Notch。通路分析显示MAPK-ERK信号传导是主要的JAG 1/Notch调节因子，这得到了51个乳腺癌细胞系中基因集富集分析的支持。根据化学筛选，激酶组小干扰RNA高通量筛选鉴定了Tribbles同源物3（TRB 3），其是MAPK-ERK的已知调节剂，是最显著的命中物之一。我们证明TRB 3是通过MAPK-ERK和TGF β途径调节Notch的主要调节因子。互补的体外和体内研究强调了TRB 3对肿瘤生长的重要性。这些数据表明TRB 3和MAPK-ERK/TGF β途径作为Notch调节剂在乳腺癌中的主导作用，确立TRB 3作为潜在的治疗靶点。
Expression of the Notch ligand Jagged 1 (JAG1) and Notch activation promote poor-prognosis in breast cancer. We used high throughput screens to identify elements responsible for Notch activation in this context. Chemical kinase inhibitor and kinase-specific small interfering RNA libraries were screened in a breast cancer cell line engineered to report Notch. Pathway analyses revealed MAPK-ERK signaling to be the predominant JAG1/Notch regulator and this was supported by gene set enrichment analyses in 51 breast cancer cell lines. In accordance with the chemical screen, kinome small interfering RNA high throughput screens identified Tribbles homolog 3 (TRB3), a known regulator of MAPK-ERK, among the most significant hits. We demonstrate that TRB3 is a master regulator of Notch through the MAPK-ERK and TGF beta pathways. Complementary in vitro and in vivo studies underscore the importance of TRB3 for tumor growth. These data demonstrate a dominant role for TRB3 and MAPK-ERK/TGF beta pathways as Notch regulators in breast cancer, establishing TRB3 as a potential therapeutic target.