Limits...
Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide.

Lee YS, Park JS, Jung SM, Kim SD, Kim JH, Lee JY, Jung KC, Mamura M, Lee S, Kim SJ, Bae YS, Park SH - EMBO Mol Med (2015)

Bottom Line: Here, we developed Smaducin-6, a novel membrane-tethered palmitic acid-conjugated Smad6-derived peptide composed of amino acids 422-441 of Smad6.Smaducin-6 interacted with Pellino-1, located in the inner membrane, thereby disrupting the formation of IRAK1-, RIP1-, IKKε-mediated TLR4 signaling complexes.Our findings provide clues to develop new peptide-based drugs to target Pellino-1 protein in TLR4 signaling pathway for the treatment of sepsis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Sungkyunkwan University, Suwon, Korea.

No MeSH data available.


Related in: MedlinePlus

Smaducin-6 is therapeutic for mice with CLP-induced sepsisSubcutaneous injection of Smaducin-6 increases the survival rate of severe CLP-induced sepsis mice (BALB/c mice). Different amounts of Smaducin-6 were subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group per experiment.Smaducin-6 treatment reduces abnormal nuclei and cell morphology observed in tissues of CLP-induced BALB/c mice, as observed by hematoxylin and eosin (H/E) staining. Data shown are representative of five independent experiments. Scale bar, 40 μm (magnification, 400×).The survival rates of severe CLP-induced sepsis mice by subcutaneously injected Smaducin-6 were compared with those by intraperitoneally injected antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin). Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. n = 10 mice per group per experiment.Therapeutic effects of Smaducin-6 alone, or antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin), or different amounts of Smaducin-6 plus antibiotics were observed in CLP mice. Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. Smaducin-6 was subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group.Comparison of mortality of severe CLP-induced sepsis mice upon subcutaneous injections of a total of 16 mg/kg Smaducin-6 or TAT-S6(422-441) peptide. n = 10 mice per group.Data information: In (A–E), PBS and scrambled peptide (Pal-Scram #1) were subcutaneously injected into CLP mice, respectively, as negative controls. Data in (A–E) were statistically analyzed by the log-rank test. ***P < 0.001, **P < 0.005, *P < 0.05 compared to vehicle control (CLP + Pal-Scram #1 or CLP + PBS).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4492818&req=5

fig04: Smaducin-6 is therapeutic for mice with CLP-induced sepsisSubcutaneous injection of Smaducin-6 increases the survival rate of severe CLP-induced sepsis mice (BALB/c mice). Different amounts of Smaducin-6 were subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group per experiment.Smaducin-6 treatment reduces abnormal nuclei and cell morphology observed in tissues of CLP-induced BALB/c mice, as observed by hematoxylin and eosin (H/E) staining. Data shown are representative of five independent experiments. Scale bar, 40 μm (magnification, 400×).The survival rates of severe CLP-induced sepsis mice by subcutaneously injected Smaducin-6 were compared with those by intraperitoneally injected antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin). Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. n = 10 mice per group per experiment.Therapeutic effects of Smaducin-6 alone, or antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin), or different amounts of Smaducin-6 plus antibiotics were observed in CLP mice. Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. Smaducin-6 was subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group.Comparison of mortality of severe CLP-induced sepsis mice upon subcutaneous injections of a total of 16 mg/kg Smaducin-6 or TAT-S6(422-441) peptide. n = 10 mice per group.Data information: In (A–E), PBS and scrambled peptide (Pal-Scram #1) were subcutaneously injected into CLP mice, respectively, as negative controls. Data in (A–E) were statistically analyzed by the log-rank test. ***P < 0.001, **P < 0.005, *P < 0.05 compared to vehicle control (CLP + Pal-Scram #1 or CLP + PBS).

Mentions: To explore the therapeutic effect of Smaducin-6 on TLR4-related inflammatory diseases, we applied cecal-ligation–puncture (CLP) to BALB/c mice (Hubbard et al, 2005). We optimized the protocol for administration of Smaducin-6: time of initiation post-CLP and the route of injection. We subcutaneously injected total 16 mg/kg of Smaducin-6 into the mice subjected to CLP at 2, 6, and 10 h post-CLP followed by three injections at 12-h intervals. Initiation at 2 h post-CLP was the most effective with 90% survival rate, and later time points reduced the efficacy; 6 h post-CLP showed 60% survival rate, and 10 h post-CLP was ineffective (Supplementary Fig S7A). The best efficacy obtained at earlier time point is consistent with the fact that any anti-inflammatory therapies against sepsis should be initiated at early time point before developing septic shock, in which hemodynamic control is the most emergent and prioritized therapy. We then compared intravenous and subcutaneous injections at 2 h post-CLP followed by three injections at 12-h intervals. Intravenous injections of Smaducin-6 were not so effective as subcutaneous injections, showing 40–60% survival rates (Supplementary Fig S7B; Fig4A). We compared the tissue biodistributions of subcutaneously or intravenously injected Smaducin-6 in mice administered with fluorescent dye TAMRA (carboxytetramethylrhodamine)-conjugated Smaducin-6 (Supplementary Table S1). After 1 h of administration, we prepared various tissues from the mice and observed TAMRA using confocal microscope. Subcutaneously injected Smaducin-6 was distributed in the spleen, lungs, liver, and kidneys, whereas it was barely detected in the heart and brain (Supplementary Fig S8A). In contrast, intravenously injected Smaducin-6 was not detected as much as subcutaneously injected Smaducin-6 in any tested tissues (Supplementary Fig S8B). Subcutaneous injection is more effective than intravenous injection presumably because of the higher biodistribution of Smaducin-6 through lymphatic vessels than blood vessels. However, it is impossible to exclude other possibilities such as decreased stability of Smaducin-6 when intravenously injected.


Inhibition of lethal inflammatory responses through the targeting of membrane-associated Toll-like receptor 4 signaling complexes with a Smad6-derived peptide.

Lee YS, Park JS, Jung SM, Kim SD, Kim JH, Lee JY, Jung KC, Mamura M, Lee S, Kim SJ, Bae YS, Park SH - EMBO Mol Med (2015)

Smaducin-6 is therapeutic for mice with CLP-induced sepsisSubcutaneous injection of Smaducin-6 increases the survival rate of severe CLP-induced sepsis mice (BALB/c mice). Different amounts of Smaducin-6 were subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group per experiment.Smaducin-6 treatment reduces abnormal nuclei and cell morphology observed in tissues of CLP-induced BALB/c mice, as observed by hematoxylin and eosin (H/E) staining. Data shown are representative of five independent experiments. Scale bar, 40 μm (magnification, 400×).The survival rates of severe CLP-induced sepsis mice by subcutaneously injected Smaducin-6 were compared with those by intraperitoneally injected antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin). Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. n = 10 mice per group per experiment.Therapeutic effects of Smaducin-6 alone, or antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin), or different amounts of Smaducin-6 plus antibiotics were observed in CLP mice. Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. Smaducin-6 was subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group.Comparison of mortality of severe CLP-induced sepsis mice upon subcutaneous injections of a total of 16 mg/kg Smaducin-6 or TAT-S6(422-441) peptide. n = 10 mice per group.Data information: In (A–E), PBS and scrambled peptide (Pal-Scram #1) were subcutaneously injected into CLP mice, respectively, as negative controls. Data in (A–E) were statistically analyzed by the log-rank test. ***P < 0.001, **P < 0.005, *P < 0.05 compared to vehicle control (CLP + Pal-Scram #1 or CLP + PBS).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4492818&req=5

fig04: Smaducin-6 is therapeutic for mice with CLP-induced sepsisSubcutaneous injection of Smaducin-6 increases the survival rate of severe CLP-induced sepsis mice (BALB/c mice). Different amounts of Smaducin-6 were subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group per experiment.Smaducin-6 treatment reduces abnormal nuclei and cell morphology observed in tissues of CLP-induced BALB/c mice, as observed by hematoxylin and eosin (H/E) staining. Data shown are representative of five independent experiments. Scale bar, 40 μm (magnification, 400×).The survival rates of severe CLP-induced sepsis mice by subcutaneously injected Smaducin-6 were compared with those by intraperitoneally injected antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin). Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. n = 10 mice per group per experiment.Therapeutic effects of Smaducin-6 alone, or antibiotics (8 mg/kg gentamycin plus 8 mg/kg cephalosporin), or different amounts of Smaducin-6 plus antibiotics were observed in CLP mice. Antibiotics were intraperitoneally injected into CLP mice at 2 h post-CLP followed by an additional injection after 12 h. Smaducin-6 was subcutaneously injected at 2 h post-CLP followed by three injections at 12-h intervals. n = 10 mice per group.Comparison of mortality of severe CLP-induced sepsis mice upon subcutaneous injections of a total of 16 mg/kg Smaducin-6 or TAT-S6(422-441) peptide. n = 10 mice per group.Data information: In (A–E), PBS and scrambled peptide (Pal-Scram #1) were subcutaneously injected into CLP mice, respectively, as negative controls. Data in (A–E) were statistically analyzed by the log-rank test. ***P < 0.001, **P < 0.005, *P < 0.05 compared to vehicle control (CLP + Pal-Scram #1 or CLP + PBS).
Mentions: To explore the therapeutic effect of Smaducin-6 on TLR4-related inflammatory diseases, we applied cecal-ligation–puncture (CLP) to BALB/c mice (Hubbard et al, 2005). We optimized the protocol for administration of Smaducin-6: time of initiation post-CLP and the route of injection. We subcutaneously injected total 16 mg/kg of Smaducin-6 into the mice subjected to CLP at 2, 6, and 10 h post-CLP followed by three injections at 12-h intervals. Initiation at 2 h post-CLP was the most effective with 90% survival rate, and later time points reduced the efficacy; 6 h post-CLP showed 60% survival rate, and 10 h post-CLP was ineffective (Supplementary Fig S7A). The best efficacy obtained at earlier time point is consistent with the fact that any anti-inflammatory therapies against sepsis should be initiated at early time point before developing septic shock, in which hemodynamic control is the most emergent and prioritized therapy. We then compared intravenous and subcutaneous injections at 2 h post-CLP followed by three injections at 12-h intervals. Intravenous injections of Smaducin-6 were not so effective as subcutaneous injections, showing 40–60% survival rates (Supplementary Fig S7B; Fig4A). We compared the tissue biodistributions of subcutaneously or intravenously injected Smaducin-6 in mice administered with fluorescent dye TAMRA (carboxytetramethylrhodamine)-conjugated Smaducin-6 (Supplementary Table S1). After 1 h of administration, we prepared various tissues from the mice and observed TAMRA using confocal microscope. Subcutaneously injected Smaducin-6 was distributed in the spleen, lungs, liver, and kidneys, whereas it was barely detected in the heart and brain (Supplementary Fig S8A). In contrast, intravenously injected Smaducin-6 was not detected as much as subcutaneously injected Smaducin-6 in any tested tissues (Supplementary Fig S8B). Subcutaneous injection is more effective than intravenous injection presumably because of the higher biodistribution of Smaducin-6 through lymphatic vessels than blood vessels. However, it is impossible to exclude other possibilities such as decreased stability of Smaducin-6 when intravenously injected.

Bottom Line: Here, we developed Smaducin-6, a novel membrane-tethered palmitic acid-conjugated Smad6-derived peptide composed of amino acids 422-441 of Smad6.Smaducin-6 interacted with Pellino-1, located in the inner membrane, thereby disrupting the formation of IRAK1-, RIP1-, IKKε-mediated TLR4 signaling complexes.Our findings provide clues to develop new peptide-based drugs to target Pellino-1 protein in TLR4 signaling pathway for the treatment of sepsis.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Sungkyunkwan University, Suwon, Korea.

No MeSH data available.


Related in: MedlinePlus