Multipathway model enables prediction of kinase inhibitor cross-talk effects on migration of Her2-overexpressing mammary epithelial cells
Multipathway model enables prediction of kinase inhibitor cross-talk effects on migration of Her2-overexpressing mammary epithelial cells
复制标题
DOI：
10.1124/mol.107.043794
复制
发表时间：
2008-06-01
影响因子：
3.6
通讯作者：
Lauffenburger, Douglas A.
中科院分区：
文献类型：
--
作者：
Kumar, Neil;Afeyan, Raffi;Lauffenburger, Douglas A.
小分子激酶抑制剂通常通过直接的“脱靶”效应或间接的“通路串扰”效应来调节靶向信号通路以外的信号通路。这两类复杂因素中的一种或两种的存在阻碍了对激酶抑制剂对药物疗效反应所涉及的细胞表型行为的影响的预测性理解。为了解决这个问题，我们提供了一个途径，了解激酶抑制剂如何调节细胞信号网络导致改变细胞表型反应，通过应用定量的，多途径的计算建模方法。我们表明，整合测量信号跨越三个关键激酶途径参与调节人类乳腺上皮细胞的迁移，ErbB系统受体激活的表皮生长因子（EGF）或heregulin（HRG）的下游，显著改善了对小分子抑制剂2-（4-吗啉基）-8-苯基-4H-1-苯并吡喃-4-酮（LY 294002）和2 '-氨基-3'-甲氧基黄酮（PD 98059）用于正常和HER 2过表达细胞。这些抑制剂主要针对抑制磷脂酰肌醇3-激酶（PI 3 K）和促分裂原活化蛋白激酶激酶（MEK），但已知表现出脱靶效应;此外，还认识到PI 3 K/Akt和MEK/细胞外信号调节激酶（Erk）途径之间的复杂串扰相互作用。我们在此观察到，用LY 294002处理减少HRG刺激的细胞的迁移，但不减少EGF刺激的细胞的迁移，尽管在两种条件下Akt磷酸化的减少水平相当，这表明靶抑制作用不能单方面预测针对细胞表型应答的功效。LY 294002处理后，Erk和p38磷酸化水平的连续测量，沿着EGF受体磷酸化水平的测量，揭示了这些非靶向途径的非预期调节。然而，当这些测量被纳入一个偏最小二乘回归模型，细胞迁移反应的治疗成功地预测。在类似地预测PD 98059对细胞迁移的治疗作用中，发现相同的多途径模型取得了类似的成功。我们的结论是，定量，多途径建模方法可以提供一个重大的进步，了解激酶抑制剂的疗效，在面对脱靶和通路串扰的影响。
Small-molecule kinase inhibitors often modulate signaling pathways other than the one targeted, whether by direct "off-target" effects or by indirect "pathway cross-talk" effects. The presence of either or both of these classes of complicating factors impedes the predictive understanding of kinase inhibitor consequences for cell phenotypic behaviors involved in drug efficacy responses. To address this problem, we offer an avenue toward comprehending how kinase inhibitor modulations of cell signaling networks lead to altered cell phenotypic responses by applying a quantitative, multipathway computational modeling approach. We show that integrating measurements of signals across three key kinase pathways involved in regulating migration of human mammary epithelial cells, downstream of ErbB system receptor activation by epidermal growth factor (EGF) or heregulin (HRG), significantly improves prediction of cell migration changes resulting from treatment with the small-molecule inhibitors 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one (LY294002) and 2'-amino-3'-methoxyflavone (PD98059) for both normal and HER2-overexpressing cells. These inhibitors are primarily directed toward inhibition of phosphatidylinositol 3-kinase (PI3K) and mitogen-activated protein kinase kinase (MEK) but are known to exhibit off-target effects; moreover, complex crosstalk interactions between the PI3K/Akt and MEK/extracellular signal-regulated kinase (Erk) pathways are also appreciated. We observe here that treatment with LY294002 reduces migration of HRG-stimulated cells but not EGF-stimulated cells, despite comparable levels of reduction of Akt phosphorylation under both conditions, demonstrating that the target inhibition effect is not unilaterally predictive of efficacy against cell phenotypic response. Consequent measurement of levels of Erk and p38 phosphorylation, along with those for EGF receptor phosphorylation, after LY294002 treatment revealed unintended modulation of these nontargeted pathways. However, when these measurements were incorporated into a partial least-squares regression model, the cell migration responses to treatment were successfully predicted. Similar success was found for the same multipathway model in analogously predicting PD98059 treatment effects on cell migration. We conclude that a quantitative, multipathway modeling approach can provide a significant advance toward comprehending kinase inhibitor efficacy in the face of off-target and pathway cross-talk effects.