Department of Drug Discovery, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, 33612, USA.
Cancer Biology Ph.D. Program, University of South Florida, Tampa, FL-33620, USA.
Chembiochem. 2023 Jun 1;24(11):e202200766. doi: 10.1002/cbic.202200766. Epub 2023 May 4.
Metastasis poses a major challenge in cancer management, including EML4-ALK-rearranged non-small cell lung cancer (NSCLC). As cell migration is a critical step during metastasis, we assessed the anti-migratory activities of several clinical ALK inhibitors in NSCLC cells and observed differential anti-migratory capabilities despite similar ALK inhibition, with brigatinib displaying superior anti-migratory effects over other ALK inhibitors. Applying an unbiased in situ mass spectrometry-based chemoproteomics approach, we determined the proteome-wide target profile of brigatinib in EML4-ALK+ NSCLC cells. Dose-dependent and cross-competitive chemoproteomics suggested MARK2 and MARK3 as relevant brigatinib kinase targets. Functional validation showed that combined pharmacological inhibition or genetic modulation of MARK2/3 inhibited cell migration. Consistently, brigatinib treatment induced inhibitory YAP1 phosphorylation downstream of MARK2/3. Collectively, our data suggest that brigatinib exhibits unusual cross-phenotype polypharmacology as, despite similar efficacy for inhibiting EML4-ALK-dependent cell proliferation as other ALK inhibitors, it more effectively prevented migration of NSCLC cells due to co-targeting of MARK2/3.
转移是癌症管理中的一个主要挑战，包括 EML4-ALK 重排的非小细胞肺癌（NSCLC）。由于细胞迁移是转移过程中的一个关键步骤，我们评估了几种临床 ALK 抑制剂在 NSCLC 细胞中的抗迁移活性，尽管 ALK 抑制相似，但观察到不同的抗迁移能力，布加替尼显示出优于其他 ALK 抑制剂的抗迁移作用。应用基于无偏置原位质谱的化学蛋白质组学方法，我们确定了布加替尼在 EML4-ALK+ NSCLC 细胞中的全蛋白质组靶标谱。剂量依赖性和交叉竞争化学蛋白质组学表明 MARK2 和 MARK3 是相关的布加替尼激酶靶标。功能验证表明，联合药理学抑制或遗传调节 MARK2/3 可抑制细胞迁移。一致地，布加替尼处理诱导了 MARK2/3 下游抑制性 YAP1 磷酸化。总之，我们的数据表明，布加替尼表现出异常的交叉表型多效性，尽管与其他 ALK 抑制剂抑制 EML4-ALK 依赖性细胞增殖的疗效相似，但由于共同靶向 MARK2/3，它更有效地阻止了 NSCLC 细胞的迁移。
