Glycogen synthase kinase 3 beta positively regulates Notch signaling in vascular smooth muscle cells: role in cell proliferation and survival

The role of glycogen synthase kinase 3 beta (GSK-3 beta) in modulating Notch control of vascular smooth muscle cell (vSMC) growth (proliferation and apoptosis) was examined in vitro under varying conditions of cyclic strain and validated in vivo following changes in medial tension and stress. Modulation of GSK-3 beta in vSMC following ectopic expression of constitutively active GSK-3 beta, siRNA knockdown and pharmacological inhibition with SB-216763 demonstrated that GSK-3 beta positively regulates Notch intracellular domain expression, CBF-1/RBP-J kappa transactivation and downstream target gene mRNA levels, while concomitantly promoting vSMC proliferation and inhibiting apoptosis. In contrast, inhibition of GSK-3 beta attenuated Notch signaling and decreased vSMC proliferation and survival. Exposure of vSMC to cyclic strain environments in vitro using both a Flexercell (TM) Tension system and a novel Sylgard (TM) phantom vessel following bare metal stent implantation revealed that cyclic strain inhibits GSK-3 beta activity independent of p42/p44 MAPK and p38 activation concomitant with reduced Notch signaling and decreased vSMC proliferation and survival. Exposure of vSMC to changes in medial strain microenvironments in vivo following carotid artery ligation revealed that enhanced GSK-3 beta activity was predominantly localized to medial and neointimal vSMC concomitant with increased Notch signaling, proliferating nuclear antigen and decreased Bax expression, respectively, as vascular remodeling progressed. GSK-3 beta is an important modulator of Notch signaling leading to altered vSMC cell growth where low strain/tension microenvironments prevail.