Low-Dose 6-Bromoindirubin-3′-oxime Induces Partial Dedifferentiation of Endothelial Cells to Promote Increased Neovascularization. (23rd May 2014)

• Record Type: Journal Article
• Authors: Kohler, Erin E.; Baruah, Jugajyoti; Urao, Norifumi; Ushio-Fukai, Masuko; Fukai, Tohru; Chatterjee, Ishita; Wary, Kishore K.
• Is Part Of: Stem cells. Volume 32:Number 6(2014:Jun.)
• Page Start: 1538
• Page End: 1552
• Publication Date: 2014-05-23
• Subjects: 6-Bromoindirubin-3′-oxime -- Dedifferentiation -- Endothelial cells -- Hind limb ischemia -- Neovascularization; Cloning -- Periodicals; Clone cells -- Periodicals; Stem cells -- Periodicals; Cell Differentiation -- Periodicals; Cell Division -- Periodicals; Clone Cells -- Periodicals; Hematopoietic Stem Cells -- Periodicals; Stem Cells -- Periodicals; 571.84
• DOI: 10.1002/stem.1658
• Languages: English
• ISSNs: 1066-5099

Abstract

Abstract : Endothelial cell (EC) dedifferentiation in relation to neovascularization is a poorly understood process. In this report, we addressed the role of Wnt signaling in the mechanisms of neovascularization in adult tissues. Here, we show that a low-dose of 6-bromoindirubin-3′-oxime (BIO), a competitive inhibitor of glycogen synthase kinase-3 β, induced the stabilization of β -catenin and its subsequent direct interaction with the transcription factor NANOG in the nucleus of ECs. This event induced loss of VE-cadherin from the adherens junctions, increased EC proliferation accompanied by asymmetric cell division (ACD), and formed cellular aggregates in hanging drop assays indicating the acquisition of a dedifferentiated state. In a chromatin immunoprecipitation assay, nuclear NANOG protein bound to the NANOG- and VEGFR2- promoters in ECs, and the addition of BIO activated the NANOG -promoter-luciferase reporter system in a cell-based assay. Consequently, NANOG -knockdown decreased BIO-induced NOTCH-1 expression, thereby decreasing cell proliferation, ACD, and neovascularization. In a Matrigel plug assay, BIO induced increased neovascularization, secondary to the presence of vascular endothelial growth factor (VEGF). Moreover, in a mouse model of hind limb ischemia, BIO augmented neovascularization that was coupled with increased expression of NOTCH-1 in ECs and increased smooth muscle α-actin + cell recruitment around the neovessels. Thus, these results demonstrate the ability of a low-dose of BIO to augment neovascularization secondary to VEGF, a process that was accompanied by a partial dedifferentiation of ECs via β -catenin and the NANOG signaling pathway. Stem Cells 2014;32:1538–1552