Anti‐angiogenic properties of a sulindac analogue

Abstract

Background and purpose: Angiopoietins (Ang) are crucial for new blood vessel formation and exert their effects by acting on the Tie2 receptor. We have recently described a sulindac analogue 2-((1E,Z)-1-benzylidene-5-bromo-2-methyl-1H-inden-3-yl)acetic acid; termed C-18 from now onwards) that inhibits Tie2 receptor activity in kinase assays in vitro. Here, we have assessed the ability of C-18 to inhibit angiogenesis-related properties of endothelial cells and tested its selectivity for the Tie2 receptor. Experimental approach: For in vitro experiments human umbilical vein endothelial cells (HUVEC) were used. Proliferation was measured using the MTT assay; migration assays were performed in a modified Boyden chamber and tube-like structure formation was determined on matrigel. The effects of C-18 in vivo were evaluated in the chicken chorioallantoic membrane (CAM). Key results: Pre-treatment of HUVEC with C-18 blocked Ang-1-stimulated migration, but also abolished vascular endothelial cell growth factor (VEGF)-and fibroblast growth factor 2-induced responses. Incubation with C-18 inhibited serum-induced proliferation in a concentration-dependent manner; C-18 was, however, without effect on Ang-1-induced survival. In addition, we observed that C-18 did not inhibit ligand-induced receptor phosphorylation of Tie2 or VEGFR2. On the other hand, C-18 blocked activation of members of the mitogen-activated protein kinase family and of the Ser/Thr kinase Akt induced by both VEGF and Ang-1. Furthermore, incubation of CAMs with C-18 led to a dose-dependent inhibition of vascular length. Conclusions and implications: C-18 did not act as a Tie2 inhibitor, as originally thought, but rather inhibited growth factorstimulated signalling pathways that regulate endothelial cell migration and potently reduces neovascularization in vivo.

4-isothiocyanate-2, 2, 6, 6-tetramethyl piperidinooxyl inhibits angiogenesis by suppressing VEGFR2 and Tie2 phosphorylation

“…Blockade of the Ang1/Tie2 and VEGF/VEGFR2 signaling pathways strengthened the anti-VEGF therapy efficacy on angiogenesis (33,34). In our study, we observed that 4-ISO-Tempo blocked the phosphorylation of Tie2 at Tyr 992 after stimulating Ang1 (200 ng/ml) for 30 min.…”

Application of magnetic force‐based cell patterning for controlling cell–cell interactions in angiogenesis

“…Angiogenesis is a tightly regulated, multi-step process that involves endothelial cell proliferation, migration, alignment, tube formation, and organization into capillary structures (Pyriochou et al, 2007). Tissue engineering seeks to provide regenerated tissue architectures in vitro and create thick, highly vascularized, multi-functional tissues capable of replicating native structures (Tsuda et al, 2007).…”

Lipopolysaccharides (LPS) Induced Angiogenesis During Chicken Embryogenesis is Abolished by Combined ETA/ETB Receptor Blockade

“…The signal for ET-1 mRNA was most prominent and the signal for ETRA was relatively low as compared to the expression of VEGFR2, a maker of angiogenesis [12][13][14]. Combined blockage of ET receptors using classical ETRA and ETRB blockers did not affect baseline angiogenesis and the mRNA expression of the angiogenesis marker VEGFR2, whereas ETRA and ETRB mRNA expression were significantly decreased after combined ETRA/ETRB blockade.…”