Kinase inhibitors are promising drugs to stabilize the endothelial barrier following inflammatory damage. However, our limited knowledge of how kinase signaling activates barrier-restorative pathways and the complexity of multi-target drugs have hindered drug discovery and repurposing efforts. Here, we apply a kinase regression approach that exploits drug polypharmacology to investigate endothelial barrier regulation. A screen of 28 kinase inhibitors identified multiple inhibitors that promote endothelial barrier integrity and revealed divergent barrier phenotypes for BCR-ABL drugs. Target deconvolution predicted 50 barrier-regulating kinases from diverse kinase families. Using gene knockdowns, we identified kinases with a role in endothelial barrier regulation and dissected different mechanisms of action of barrier-protective kinase inhibitors. These results demonstrate the importance of polypharmacology in the endothelial barrier phenotype of kinase inhibitors and provide promising new leads for barrier-strengthening therapies.

激酶抑制剂是有前途的药物，可在炎症损伤后稳定内皮屏障。然而，我们对激酶信号如何激活屏障修复途径的了解有限以及多靶点药物的复杂性阻碍了药物发现和再利用工作。在这里，我们应用一种激酶回归方法，利用药物多药理学来研究内皮屏障调节。对 28 种激酶抑制剂的筛选确定了多种促进内皮屏障完整性的抑制剂，并揭示了 BCR-ABL 药物的不同屏障表型。目标去卷积预测了来自不同激酶家族的 50 种屏障调节激酶。使用基因敲低，我们鉴定了在内皮屏障调节中起作用的激酶，并剖析了屏障保护性激酶抑制剂的不同作用机制。