Namjoo Mohadeseh, Ghafouri Hossein, Assareh Elham, Aref Amir Reza, Mostafavi Ebrahim, Hamrahi Mohsen Ali, Balalaie Saeed, Broussy Sylvain, Asghari S Mohsen
Department of Biology, University Campus II, University of Guilan, Rasht P.O. Box 14155-6619, Iran.
Department of Biology, Faculty of Sciences, University of Guilan, Rasht P.O. Box 14155-6619, Iran.
Pharmaceuticals (Basel). 2023 Jun 20;16(6):906. doi: 10.3390/ph16060906.
Vascular endothelial growth factor receptor 2 (VEGFR2) mediates VEGFA signaling mainly through the PI3K/AKT/mTOR and PLCγ/ERK1/2 pathways. Here we unveil a peptidomimetic (VGB3) based on the interaction between VEGFB and VEGFR1 that unexpectedly binds and neutralizes VEGFR2. Investigation of the cyclic and linear structures of VGB3 (named C-VGB3 and L-VGB3, respectively) using receptor binding and cell proliferation assays, molecular docking, and evaluation of antiangiogenic and antitumor activities in the 4T1 mouse mammary carcinoma tumor (MCT) model showed that loop formation is essential for peptide functionality. C-VGB3 inhibited proliferation and tubulogenesis of human umbilical vein endothelial cells (HUVECs), accounting for the abrogation of VEGFR2, p-VEGFR2 and, subsequently, PI3K/AKT/mTOR and PLCγ/ERK1/2 pathways. In 4T1 MCT cells, C-VGB3 inhibited cell proliferation, VEGFR2 expression and phosphorylation, the PI3K/AKT/mTOR pathway, FAK/Paxillin, and the epithelial-to-mesenchymal transition cascade. The apoptotic effects of C-VGB3 on HUVE and 4T1 MCT cells were inferred from annexin-PI and TUNEL staining and activation of P53, caspase-3, caspase-7, and PARP1, which mechanistically occurred through the intrinsic pathway mediated by Bcl2 family members, cytochrome c, Apaf-1 and caspase-9, and extrinsic pathway via death receptors and caspase-8. These data indicate that binding regions shared by VEGF family members may be important in developing novel pan-VEGFR inhibitors that are highly relevant in the pathogenesis of angiogenesis-related diseases.
血管内皮生长因子受体2（VEGFR2）主要通过PI3K/AKT/mTOR和PLCγ/ERK1/2途径介导VEGFA信号传导。在此，我们基于VEGFB与VEGFR1之间的相互作用揭示了一种拟肽（VGB3），其意外地结合并中和了VEGFR2。使用受体结合和细胞增殖试验、分子对接以及在4T1小鼠乳腺癌肿瘤（MCT）模型中评估抗血管生成和抗肿瘤活性，对VGB3的环状和线性结构（分别命名为C-VGB3和L-VGB3）进行研究，结果表明环的形成对于肽的功能至关重要。C-VGB3抑制人脐静脉内皮细胞（HUVECs）的增殖和管状形成，这是VEGFR2、p-VEGFR2以及随后的PI3K/AKT/mTOR和PLCγ/ERK1/2途径被废除的原因。在4T1 MCT细胞中，C-VGB3抑制细胞增殖、VEGFR2表达和磷酸化、PI3K/AKT/mTOR途径、FAK/桩蛋白以及上皮-间质转化级联反应。通过膜联蛋白-PI和TUNEL染色以及P53、半胱天冬酶-3、半胱天冬酶-7和PARP1的激活推断出C-VGB3对HUVE和4T1 MCT细胞的凋亡作用，其机制是通过由Bcl2家族成员、细胞色素c、凋亡蛋白酶激活因子-1和半胱天冬酶-9介导的内源性途径以及通过死亡受体和半胱天冬酶-8的外源性途径发生的。这些数据表明，VEGF家族成员共有的结合区域在开发新型泛VEGFR抑制剂中可能很重要，这些抑制剂在血管生成相关疾病的发病机制中具有高度相关性。
