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GSK3beta Inhibitor Peptide Protects Mice from LPS-induced Endotoxin Shock.

Ko R, Jang HD, Lee SY - Immune Netw (2010)

Bottom Line: We first demonstrate its effects on LPS-stimulated pro-inflammatory cytokine production including interleukin (IL)-6 and IL-12p40.Collectively, we present a rational strategy for the development of a therapeutic GSK3i peptide.This peptide may serve as a novel template for the design of non-ATP competitive GSK3 inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Division of Life and Pharmaceutical Sciences, Center for Cell Signaling & Drug Discovery Research, Ewha Womans University, Seoul 120-750, Korea.

ABSTRACT

Background: Glycogen synthase kinase 3beta (GSK3beta) is a ubiquitous serine/threonine kinase that is regulated by serine phosphorylation at 9. Recent studies have reported the beneficial effects of a number of the pharmacological GSK3beta inhibitors in rodent models of septic shock. Since most of the GSK3beta inhibitors are targeted at the ATP-binding site, which is highly conserved among diverse protein kinases, the development of novel non-ATP competitive GSK3beta inhibitors is needed.

Methods: Based on the unique phosphorylation motif of GSK3beta, we designed and generated a novel class of GSK3beta inhibitor (GSK3i) peptides. In addition, we investigated the effects of a GSK3i peptide on lipopolysaccharide (LPS)-stimulated cytokine production and septic shock. Mice were intraperitoneally injected with GSK3i peptide and monitored over a 7-day period for survival.

Results: We first demonstrate its effects on LPS-stimulated pro-inflammatory cytokine production including interleukin (IL)-6 and IL-12p40. LPS-induced IL-6 and IL-12p40 production in macrophages was suppressed when macrophages were treated with the GSKi peptide. Administration of the GSK3i peptide potently suppressed LPS-mediated endotoxin shock.

Conclusion: Collectively, we present a rational strategy for the development of a therapeutic GSK3i peptide. This peptide may serve as a novel template for the design of non-ATP competitive GSK3 inhibitors.

No MeSH data available.


Related in: MedlinePlus

The GSKi peptide decreased pro-inflammatory cytokines production after LPS stimulation. BMDMs were pre-incubated for 2 hours with medium only and either 10 µM SB216763 or 5 µM GSK3i peptide, and then stimulated with 1 µg/ml LPS for 20 hours. Cell-free supernatants were analyzed by ELISA for production of pro-inflammatory cytokines; IL-6 (A) or IL-12p40 (B). Data represent mean±s.d. and are representative of at least three experiments.
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Figure 2: The GSKi peptide decreased pro-inflammatory cytokines production after LPS stimulation. BMDMs were pre-incubated for 2 hours with medium only and either 10 µM SB216763 or 5 µM GSK3i peptide, and then stimulated with 1 µg/ml LPS for 20 hours. Cell-free supernatants were analyzed by ELISA for production of pro-inflammatory cytokines; IL-6 (A) or IL-12p40 (B). Data represent mean±s.d. and are representative of at least three experiments.

Mentions: We designed a cell-permeable GSKi peptide spanning the serine 9 phosphorylation motif of GSK3β that was fused with recently characterized cell-permeable sequences derived from the human transcription factor Hph-1 (Fig. 1) (29,30). Since GSK3β is known key regulator of pro-inflammatory cytokine production (16), we examined the ability of the GSKi peptide to regulate cytokine production in response to LPS stimulation. BMDMs from male 6~8 week-old mice were pre-incubated for 2 hours with either 5 µM GSKi peptide or 10 µM SB216763 as a positive control, and then the cells were stimulated with 1 µg/ml LPS for 20 hours. The control peptide containing the cell-permeable sequences only did not affect cytokine production stimulated by LPS (data not shown). As shown in Fig. 2, the presence of the GSK3i peptide was shown to attenuate pro-inflammatory cytokine production; IL-6 and IL-12p40. These inhibitory effects were comparable to that of SB216763 which is a well-characterized pharmacological inhibitor of GSK3. These results demonstrate that the GSKi peptide can regulate LPS-mediated pro-inflammatory cytokine production.


GSK3beta Inhibitor Peptide Protects Mice from LPS-induced Endotoxin Shock.

Ko R, Jang HD, Lee SY - Immune Netw (2010)

The GSKi peptide decreased pro-inflammatory cytokines production after LPS stimulation. BMDMs were pre-incubated for 2 hours with medium only and either 10 µM SB216763 or 5 µM GSK3i peptide, and then stimulated with 1 µg/ml LPS for 20 hours. Cell-free supernatants were analyzed by ELISA for production of pro-inflammatory cytokines; IL-6 (A) or IL-12p40 (B). Data represent mean±s.d. and are representative of at least three experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC2902676&req=5

Figure 2: The GSKi peptide decreased pro-inflammatory cytokines production after LPS stimulation. BMDMs were pre-incubated for 2 hours with medium only and either 10 µM SB216763 or 5 µM GSK3i peptide, and then stimulated with 1 µg/ml LPS for 20 hours. Cell-free supernatants were analyzed by ELISA for production of pro-inflammatory cytokines; IL-6 (A) or IL-12p40 (B). Data represent mean±s.d. and are representative of at least three experiments.
Mentions: We designed a cell-permeable GSKi peptide spanning the serine 9 phosphorylation motif of GSK3β that was fused with recently characterized cell-permeable sequences derived from the human transcription factor Hph-1 (Fig. 1) (29,30). Since GSK3β is known key regulator of pro-inflammatory cytokine production (16), we examined the ability of the GSKi peptide to regulate cytokine production in response to LPS stimulation. BMDMs from male 6~8 week-old mice were pre-incubated for 2 hours with either 5 µM GSKi peptide or 10 µM SB216763 as a positive control, and then the cells were stimulated with 1 µg/ml LPS for 20 hours. The control peptide containing the cell-permeable sequences only did not affect cytokine production stimulated by LPS (data not shown). As shown in Fig. 2, the presence of the GSK3i peptide was shown to attenuate pro-inflammatory cytokine production; IL-6 and IL-12p40. These inhibitory effects were comparable to that of SB216763 which is a well-characterized pharmacological inhibitor of GSK3. These results demonstrate that the GSKi peptide can regulate LPS-mediated pro-inflammatory cytokine production.

Bottom Line: We first demonstrate its effects on LPS-stimulated pro-inflammatory cytokine production including interleukin (IL)-6 and IL-12p40.Collectively, we present a rational strategy for the development of a therapeutic GSK3i peptide.This peptide may serve as a novel template for the design of non-ATP competitive GSK3 inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Division of Life and Pharmaceutical Sciences, Center for Cell Signaling & Drug Discovery Research, Ewha Womans University, Seoul 120-750, Korea.

ABSTRACT

Background: Glycogen synthase kinase 3beta (GSK3beta) is a ubiquitous serine/threonine kinase that is regulated by serine phosphorylation at 9. Recent studies have reported the beneficial effects of a number of the pharmacological GSK3beta inhibitors in rodent models of septic shock. Since most of the GSK3beta inhibitors are targeted at the ATP-binding site, which is highly conserved among diverse protein kinases, the development of novel non-ATP competitive GSK3beta inhibitors is needed.

Methods: Based on the unique phosphorylation motif of GSK3beta, we designed and generated a novel class of GSK3beta inhibitor (GSK3i) peptides. In addition, we investigated the effects of a GSK3i peptide on lipopolysaccharide (LPS)-stimulated cytokine production and septic shock. Mice were intraperitoneally injected with GSK3i peptide and monitored over a 7-day period for survival.

Results: We first demonstrate its effects on LPS-stimulated pro-inflammatory cytokine production including interleukin (IL)-6 and IL-12p40. LPS-induced IL-6 and IL-12p40 production in macrophages was suppressed when macrophages were treated with the GSKi peptide. Administration of the GSK3i peptide potently suppressed LPS-mediated endotoxin shock.

Conclusion: Collectively, we present a rational strategy for the development of a therapeutic GSK3i peptide. This peptide may serve as a novel template for the design of non-ATP competitive GSK3 inhibitors.

No MeSH data available.


Related in: MedlinePlus