Tideglusib, an allosteric inhibitor of Glycogen Synthase Kinase (GSK)‐3β increases Angiogenic activities of Endothelial Cells

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RationaleThe regulation of blood vessel growth is of paramount importance in ischemic cardiovascular and peripheral limb diseases. However, there is no effective pharmacotherapy to induce rapid revascularization of ischemic tissues.GoalTo address if newly discovered allosteric inhibitor of GSK‐3β enzyme, Tideglusib (TDG) regulates angiogenic phenotypes of endothelial cells.Methods and ResultsThe treatment of cultured ECs with Wnt3a or TDG stabilized β‐catenin polypeptide species in a time dependent manner. Quantitative RT‐PCR showed increased Cyclin‐D1, Nanog, c‐myc and Vascular Endothelial Growth Factor Receptor‐2(VEGFR‐2) transcripts. Accordingly, TDG increased expression of not only transcription factor Nanog, but also Cyclin‐D1 and VEGFR‐2 proteins. Microscopic analyses of ECs stained with anti‐VE‐cadherin antibody provided evidence of the disappearance of VE‐cadherin from the adherens junctions, while increased accumulation of β‐catenin in the nucleus. Pretreatment of ECs with TDG followed by Propidium iodide (PI) and Annexin‐V labeling showed increased survival and decreased cell death (p<0.05). Importantly, TDG treatment and the addition of 5‐bromo‐2′‐deoxyuridine (BrdU) followed by microscopy showed significant increase in the incorporation of BrdU in ECs. In addition, in a scratch assay and Boyden‐chamber cell migration experiments indicated the ability of TDG to induce migration of ECs. Furthermore, in an aortic ring assay TDG increased angiogenesis significantly.SummaryThus, for the first time, we show that TDG up‐regulates expression of Nanog, Cyclin‐D1 and VEGFR‐2 to augment neovessel formation, a critical biological process required for re‐establishing blood flow to the ischemic tissues.