Zibrova Darya, Ernst Thomas, Hochhaus Andreas, Heller Regine
Center for Molecular Biomedicine, Institute of Molecular Cell Biology, Jena University Hospital, Hans-Knöll-Straße 2, 07745, Jena, Germany.
Department of Hematology and Oncology, Jena University Hospital, Jena, Germany.
Mol Cell Biochem. 2025 Mar;480(3):1627-1643. doi: 10.1007/s11010-024-05070-5. Epub 2024 Jul 15.
:ABL1 inhibitors, the treatment of choice for the majority of patients with chronic myeloid leukaemia (CML), can cause vascular side effects that vary between agents. The exact underlying mechanisms are still poorly understood, but the vascular endothelium has been proposed as a site of origin. The present study investigates the effects of three BCR::ABL1 inhibitors, ponatinib, nilotinib and imatinib, on angiogenesis and signalling in human endothelial cells in response to vascular endothelial growth factor (VEGF). The experiments were performed in endothelial cells isolated from human umbilical veins. After exposure to imatinib, ponatinib and nilotinib, the angiogenic capacity of endothelial cells was assessed in spheroid assays. VEGF-induced signalling pathways were examined in Western blotting experiments using different specific antibodies. RNAi technology was used to downregulate proteins of interest. Intracellular cGMP levels were measured by ELISA. Imatinib had no effect on endothelial function. Ponatinib inhibited VEGF-induced sprouting, while nilotinib increased spontaneous and VEGF-stimulated angiogenesis. These effects did not involve wild-type ABL1 or ABL2, as siRNA-mediated knockdown of these kinases did not affect angiogenesis and VEGF signalling. Consistent with their effects on sprouting, ponatinib and nilotinib affected angiogenic pathways in opposite directions. While ponatinib inhibited VEGF-induced signalling and cGMP formation, nilotinib activated angiogenic signalling, in particular phosphorylation of extracellular signal-regulated kinase 1/2 (Erk1/2). The latter occurred in an epidermal growth factor receptor (EGFR)-dependent manner possibly via suppressing Fyn-related kinase (FRK), a negative regulator of EGFR signalling. Both, pharmacological inhibition of Erk1/2 or EGFR suppressed nilotinib-induced angiogenic sprouting. These results support the notion that the vascular endothelium is a site of action of BCR::ABL1 inhibitors from which side effects may arise, and that the different vascular toxicity profiles of BCR::ABL1 inhibitors may be due to their different actions at the molecular level. In addition, the as yet unknown pro-angiogenic effect of nilotinib should be considered in the treatment of patients with comorbidities associated with pathological angiogenesis, such as ocular disease, arthritis or obesity.
:ABL1抑制剂是大多数慢性髓性白血病（CML）患者的首选治疗药物，可引起不同药物之间存在差异的血管副作用。确切的潜在机制仍知之甚少，但血管内皮已被认为是起源部位。本研究调查了三种BCR::ABL1抑制剂波纳替尼、尼洛替尼和伊马替尼对人内皮细胞中血管生成和血管内皮生长因子（VEGF）信号传导的影响。实验在从人脐静脉分离的内皮细胞中进行。在暴露于伊马替尼、波纳替尼和尼洛替尼后，通过球体试验评估内皮细胞的血管生成能力。使用不同的特异性抗体，通过蛋白质印迹实验检测VEGF诱导的信号通路。RNA干扰技术用于下调感兴趣的蛋白质。通过酶联免疫吸附测定法测量细胞内cGMP水平。伊马替尼对内皮功能无影响。波纳替尼抑制VEGF诱导的芽生，而尼洛替尼增加自发的和VEGF刺激的血管生成。这些作用不涉及野生型ABL1或ABL2，因为小干扰RNA介导的这些激酶的敲低不影响血管生成和VEGF信号传导。与它们对芽生的作用一致，波纳替尼和尼洛替尼以相反的方向影响血管生成途径。虽然波纳替尼抑制VEGF诱导的信号传导和cGMP形成，但尼洛替尼激活血管生成信号传导，特别是细胞外信号调节激酶1/2（Erk1/2）的磷酸化。后者以表皮生长因子受体（EGFR）依赖性方式发生，可能是通过抑制Fyn相关激酶（FRK），EGFR信号传导的负调节因子。对Erk1/2或EGFR的药理学抑制均抑制了尼洛替尼诱导的血管生成芽生。这些结果支持这样的观点：血管内皮是BCR::ABL1抑制剂的作用位点，副作用可能由此产生，并且BCR::ABL1抑制剂不同的血管毒性特征可能归因于它们在分子水平上的不同作用。此外，在治疗与病理性血管生成相关的合并症患者，如眼部疾病、关节炎或肥胖症患者时，应考虑尼洛替尼尚未明确的促血管生成作用。
