Abstract 20373: Inhibition of Glycogen Synthase Kinase-3β Amplifies β-catenin Signaling to Promote Dedifferentiation of Endothelial Cells to Improve Neovessel Formation

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Kishore K Wary ◽  
Erin E Kohler
Keyword(s):  
Endothelial Cells ◽  
Glycogen Synthase ◽  
Glycogen Synthase Kinase ◽  
Nuclear Accumulation ◽  
Luciferase Reporter ◽  
Reporter System ◽  
Notch 1 ◽  
Matrigel Plug ◽  
Gsk 3Β

Rationale: Wnt signaling plays a key role in the development and the regeneration of neovessels, however, the underlying mechanisms of these processes are not clearly understood. Goal: To elucidate the mechanism of dedifferentiation of venous endothelial cells (VECs) to arterial (A)ECs. Methods and Results: Herein we demonstrate that 6-bromoindirubin-3'-oxime (BIO), an inhibitor of Glycogen Synthase Kinase (GSK)-3β elicits the stabilization and the nuclear accumulation of β-catenin, thereby its direct interaction with the transcription factor NANOG in the nucleus of ECs. In a ChiP experiment, NANOG protein bound to the NANOG-, BRACHYURY-, CD133- and VEGFR2 -promoters in these cells. In a cell-based assay, BIO induced the activation of the NANOG -promoter-luciferase reporter system. Accordingly, stimulation of VECs with BIO increased Notch-1, Ephrin-B2 and Hey2 expressions, indicating dedifferentiation these cells to AECs. While NANOG-depletion decreased BIO-induced NOTCH-1 expression and neovascularization. BIO increased Matrigel plug neovessel formation in nude mice. Immunostaining of Matrigel plug sections prepared from these mice showed increased expression of Notch-1, Ephrin-B2 and Hey2, indicating dedifferentiation of a subset of VECs into AECs. In a mouse model of hind limb ischemia, BIO induced the incorporation of VECs into collateral neovessels as characterized by Ephrin-B2- and Hey2-positivities, and smooth muscle cell recruitment. Summary: Thus, we show that a subset of mature VECs may retain the ability to dedifferentiate into AECs in vivo and suggest critical roles of GSK-3β inhibition in this process.

scholarly journals GSK-3β in Pancreatic Cancer: Spotlight on 9-ING-41, Its Therapeutic Potential and Immune Modulatory Properties

Biology ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 610
Author(s):  
Robin Park ◽  
Andrew L. Coveler ◽  
Ludimila Cavalcante ◽  
Anwaar Saeed
Keyword(s):  
Pancreatic Cancer ◽  
Therapeutic Potential ◽  
Glycogen Synthase ◽  
Glycogen Synthase Kinase ◽  
Glycogen Synthase Kinase 3 ◽  
In Vivo Models ◽  
Gsk 3Β ◽  
Early Phase Clinical Trials

Glycogen synthase kinase-3 beta is a ubiquitously and constitutively expressed molecule with pleiotropic function. It acts as a protooncogene in the development of several solid tumors including pancreatic cancer through its involvement in various cellular processes including cell proliferation, survival, invasion and metastasis, as well as autophagy. Furthermore, the level of aberrant glycogen synthase kinase-3 beta expression in the nucleus is inversely correlated with tumor differentiation and survival in both in vitro and in vivo models of pancreatic cancer. Small molecule inhibitors of glycogen synthase kinase-3 beta have demonstrated therapeutic potential in pre-clinical models and are currently being evaluated in early phase clinical trials involving pancreatic cancer patients with interim results showing favorable results. Moreover, recent studies support a rationale for the combination of glycogen synthase kinase-3 beta inhibitors with chemotherapy and immunotherapy, warranting the evaluation of novel combination regimens in the future.

scholarly journals Anesthetic Propofol Causes Glycogen Synthase Kinase-3β-regulated Lysosomal/Mitochondrial Apoptosis in Macrophages

2012 ◽  
Vol 116 (4) ◽  
pp. 868-881 ◽  
Author(s):  
Chung-Hsi Hsing ◽  
Yu-Hong Chen ◽  
Chia-Ling Chen ◽  
Wei-Ching Huang ◽  
Ming-Chung Lin ◽  
...  
Keyword(s):  
Glycogen Synthase ◽  
Membrane Permeabilization ◽  
Lysosomal Membrane ◽  
Glycogen Synthase Kinase ◽  
Human Neutrophils ◽  
Mitochondrial Apoptosis ◽  
Lysosomal Membrane Permeabilization ◽  
Gsk 3Β

Background Overdose propofol treatment with a prolong time causes injury to multiple cell types; however, its molecular mechanisms remain unclear. Activation of glycogen synthase kinase (GSK)-3β is proapoptotic under death stimuli. The authors therefore hypothesize that propofol overdose induces macrophage apoptosis through GSK-3β. Methods Phagocytic analysis by uptake of Staphylococcus aureus showed the effects of propofol overdose on murine macrophages RAW264.7 and BV2 and primary human neutrophils in vitro. The authors further investigated cell apoptosis in vitro and in vivo, lysosomal membrane permeabilization, and the loss of mitochondrial transmembrane potential (MTP) by propidium iodide, annexin V, acridine orange, and rhodamine 123 staining, respectively. Protein analysis identified activation of apoptotic signals, and pharmacologic inhibition and genetic knockdown using lentiviral-based short hairpin RNA were further used to clarify their roles. Results A high dose of propofol caused phagocytic inhibition and apoptosis in vitro for 24 h (25 μg/ml, in triplicate) and in vivo for 6 h (10 mg/kg/h, n = 5 for each group). Propofol induced lysosomal membrane permeabilization and MTP loss while stabilizing MTP and inhibiting caspase protected cells from mitochondrial apoptosis. Lysosomal cathepsin B was required for propofol-induced lysosomal membrane permeabilization, MTP loss, and apoptosis. Propofol decreased antiapoptotic Bcl-2 family proteins and then caused proapoptotic Bcl-2-associated X protein (Bax) activation. Propofol-activated GSK-3β and inhibiting GSK-3β prevented Mcl-1 destabilization, MTP loss, and lysosomal/mitochondrial apoptosis. Forced expression of Mcl-1 prevented the apoptotic effects of propofol. Decreased Akt was important for GSK-3β activation caused by propofol. Conclusions These results suggest an essential role of GSK-3β in propofol-induced lysosomal/mitochondrial apoptosis.

scholarly journals Glycogen synthase kinase-3-mediated phosphorylation of serine 73 targets sterol response element binding protein-1c (SREBP-1c) for proteasomal degradation

2016 ◽  
Vol 36 (1) ◽  
Author(s):  
Qingming Dong ◽  
Francesco Giorgianni ◽  
Sarka Beranova-Giorgianni ◽  
Xiong Deng ◽  
Robert N. O'Meally ◽  
...  
Keyword(s):  
Functional Role ◽  
Glycogen Synthase ◽  
Proteasomal Degradation ◽  
Glycogen Synthase Kinase ◽  
Glycogen Synthase Kinase 3 ◽  
Response Element ◽  
Element Binding Protein ◽  
Gsk 3Β

We have identified Serine 73 as a novel GSK-3β site on SREBP-1c that alters its affinity for SCAP, and proteasomal degradation. Phosphorylation of Serine 73 by GSK-3β during starvation (insulin-depleted stat) may lead to lower levels of SREBP-1c; conversely, de-phosphorylation of this site may be involved in stabilizing SREBP-1c by insulin (by blocking GSK-3β action). A functional role of this site needs to be corroborated in vivo.

Synthesis of benzimidazole-based 1,3,4-oxadiazole-1,2,3-triazole conjugates as glycogen synthase kinase-3β inhibitors with antidepressant activity in in vivo models

RSC Advances ◽  
2016 ◽  
Vol 6 (49) ◽  
pp. 43345-43355 ◽  
Author(s):  
Mushtaq A. Tantray ◽  
Imran Khan ◽  
Hinna Hamid ◽  
Mohammad Sarwar Alam ◽  
Abhijeet Dhulap ◽  
...  
Keyword(s):  
Glycogen Synthase ◽  
Antidepressant Activity ◽  
Glycogen Synthase Kinase ◽  
Glycogen Synthase Kinase 3Β ◽  
In Vivo Models ◽  
Gsk 3Β

Synthesized benzimidazole based 1,3,4-oxadiazole-1,2,3-triazole conjugates were found to inhibit GSK-3β activityin vitroand exhibit antidepressant-like activity inin vivostudies.

scholarly journals Measurement of Effector Protein Injection by Type III and Type IV Secretion Systems by Using a 13-Residue Phosphorylatable Glycogen Synthase Kinase Tag

2006 ◽  
Vol 74 (10) ◽  
pp. 5645-5657 ◽  
Author(s):  
Julie Torruellas Garcia ◽  
Franco Ferracci ◽  
Michael W. Jackson ◽  
Sabrina S. Joseph ◽  
Isabelle Pattis ◽  
...  
Keyword(s):  
Glycogen Synthase ◽  
Eukaryotic Cell ◽  
Glycogen Synthase Kinase ◽  
Cell Protein ◽  
Secretion Systems ◽  
Reporter System ◽  
Simple Method ◽  
Type Iii ◽  
Gsk 3Β ◽  
Peptide Tag

ABSTRACT Numerous bacterial pathogens use type III secretion systems (T3SSs) or T4SSs to inject or translocate virulence proteins into eukaryotic cells. Several different reporter systems have been developed to measure the translocation of these proteins. In this study, a peptide tag-based reporter system was developed and used to monitor the injection of T3S and T4S substrates. The glycogen synthase kinase (GSK) tag is a 13-residue phosphorylatable peptide tag derived from the human GSK-3β kinase. Translocation of a GSK-tagged protein into a eukaryotic cell results in host cell protein kinase-dependent phosphorylation of the tag, which can be detected with phosphospecific GSK-3β antibodies. A series of expression plasmids encoding Yop-GSK fusion proteins were constructed to evaluate the ability of the GSK tag to measure the injection of Yops by the Yersinia pestis T3SS. GSK-tagged YopE, YopH, LcrQ, YopK, YopN, and YopJ were efficiently phosphorylated when translocated into HeLa cells. Similarly, the injection of GSK-CagA by the Helicobacter pylori T4SS into different cell types was measured via phosphorylation of the GSK tag. The GSK tag provides a simple method to monitor the translocation of T3S and T4S substrates.

scholarly journals Regulation of Glycogen Synthase Kinase 3β and Downstream Wnt Signaling by Axin

1999 ◽  
Vol 19 (10) ◽  
pp. 7147-7157 ◽  
Author(s):  
Chester M. Hedgepeth ◽  
Matthew A. Deardorff ◽  
Kathleen Rankin ◽  
Peter S. Klein
Keyword(s):  
Wnt Signaling ◽  
Glycogen Synthase ◽  
Glycogen Synthase Kinase ◽  
Axis Formation ◽  
Glycogen Synthase Kinase 3Β ◽  
Adenomatous Polyposis Coli Protein ◽  
Interaction Domain ◽  
Regulatory Domains ◽  
Gsk 3Β

ABSTRACT Axin is a recently identified protein encoded by thefused locus in mice that is required for normal vertebrate axis formation. We have defined a 25-amino-acid sequence in axin that comprises the glycogen synthase kinase 3β (GSK-3β) interaction domain (GID). In contrast to full-length axin, which has been shown to antagonize Wnt signaling, the GID inhibits GSK-3β in vivo and activates Wnt signaling. Similarly, mutants of axin lacking key regulatory domains such as the RGS domain, which is required for interaction with the adenomatous polyposis coli protein, bind and inhibit GSK-3β in vivo, suggesting that these domains are critical for proper regulation of GSK-3β activity. We have identified a novel self-interaction domain in axin and have shown that formation of an axin regulatory complex in vivo is critical for axis formation and GSK-3β activity. Based on these data, we propose that the axin complex may directly regulate GSK-3β enzymatic activity in vivo. These observations also demonstrate that alternative inhibitors of GSK-3β can mimic the effect of lithium in developingXenopus embryos.

Sign in / Sign up

Export Citation Format

Share Document