Target Identification in Small Cell Lung Cancer via Integrated Phenotypic Screening and Activity-Based Protein Profiling.
Target Identification in Small Cell Lung Cancer via Integrated Phenotypic Screening and Activity-Based Protein Profiling.
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DOI：
10.1158/1535-7163.mct-15-0444
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发表时间：
2016-02
影响因子：
5.7
通讯作者：
Haura EB
中科院分区：
文献类型：
--
作者：
Li J;Fang B;Kinose F;Bai Y;Kim JY;Chen YA;Rix U;Koomen JM;Haura EB
为了克服在小细胞肺癌（SCLC）中识别关键激酶的障碍，我们将激酶抑制剂的靶标不确定表型筛选与基于活性的蛋白质谱分析（ABPP）的靶标识别结合起来，其中使用了去硫生物素- atp探针。我们使用235个小分子激酶抑制剂的化学文库筛选了21个已知c-MYC扩增状态的SCLC细胞系，以观察其活力的变化。ABPP对一个筛选命中的化合物进行了询问，通过这种方法，我们重新确定了极光激酶B是myc扩增的SCLC细胞中的关键激酶。接下来，我们将该平台扩展到在缺乏c-MYC扩增的SCLC细胞系中具有活性的第二种化合物，并鉴定了tank结合激酶1 (TBK1)，这种激酶影响缺乏MYC扩增的SCLC细胞的细胞活力、polo样激酶1信号传导、G2/M阻滞和凋亡。这些结果表明，表型筛选结合基于活性的蛋白质谱分析可以识别关键的疾病驱动因素，这表明这种结合了新的化学探针和疾病细胞筛选的方法具有识别其他癌症类型中其他重要靶点的潜力。
To overcome hurdles in identifying key kinases in small cell lung cancer (SCLC), we integrated a target-agnostic phenotypic screen of kinase inhibitors with target identification using activity-based protein profiling (ABPP) in which a desthiobiotin-ATP probe was used. We screened 21 SCLC cell lines with known c-MYC amplification status for alterations in viability using a chemical library of 235 small molecule kinase inhibitors. One screen hit compound was interrogated with ABPP, and through this approach we re-identified aurora kinase B as a critical kinase in MYC-amplified SCLC cells. We next extended the platform to a second compound that had activity in SCLC cell lines lacking c-MYC amplification and identified TANK-binding kinase 1 (TBK1), a kinase that affects cell viability, polo-like kinase-1 signaling, G2/M arrest, and apoptosis in SCLC cells lacking MYC amplification. These results demonstrate that phenotypic screening combined with activity-based protein profiling can identify key disease drivers, suggesting that this approach, which combines new chemical probes and disease cell screens, has the potential to identify other important targets in other cancer types.