Abstract 16804: Tideglusib, a Novel Allosteric Inhibitor of Glycogen Synthase Kinase-3-β Mimics Canonical Wnt Signaling
Rationale: As Glycogen Synthase Kinase-3 (GSK-3)β enzyme regulates β-catenin stability, we addressed the ability of Tideglusib, a novel allosteric inhibitor of GSK-3β, to activate canonical Wnt signaling in microvascular endothelial cells (ECs).
Methods and Results: The treatment of primary ECs with Wnt3a or Tideglusib stabilized β-catenin polypeptide species in a dose-dependent manner. A time-course analyses showed increased in phosphorylated form of GSK-3β-S9, thereby, increased stabilization of β-catenin within 3-4 hours in response to Tideglusib. The addition of Tideglusib induced robust expression of not only transcription factor Nanog, but also Cyclin-D1 and Vascular Endothelial Growth Factor Receptor-2 proteins. Microscopic analyses demonstrated disappearance of VE-cadherin from the adherens junctions, while increased accumulation of β-catenin in the nucleus. Moreover, Fluorescence-activated cell sorting (FACS) of Propidium iodide and Annexin-V labeled ECs previously treated with Tideglusib showed increased survival and decreased cell death (p<0.01 vs control groups). Accordingly, Tideglusib stimulation and the addition of 5-bromo-2'-deoxyuridine (BrdU) followed by microscopy showed increased BrdU incorporation in ECs. Furthermore, scratch wound closer and Boyden-chamber experiments provided evidence that Tideglusib induced migration of ECs in culture. Ongoing experiments are designed to address the toxicity and efficacy of Tideglusib in vivo.
Summary: For the first time, these data indicate the capacity of nanomolar concentration of Tideglusib to mimic canonical Wnt signaling to induce dedifferentiation of ECs accompanied by robust expression of transcription factor Nanog. Thus, we propose that Tideglusib could become useful in reactivating post-mitotic cells in ischemic heart and vascular tissues to replace damaged and lost cells in situ.
Author Disclosures: J. Baruah: None. K. Wary: None.
This research has received full or partial funding support from the American Heart Association
- © 2014 by American Heart Association, Inc.