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Inhibitory Effects of KP-A159, a Thiazolopyridine Derivative, on Osteoclast Differentiation, Function, and Inflammatory Bone Loss via Suppression of RANKL-Induced MAP Kinase Signaling Pathway.

Ihn HJ, Lee D, Lee T, Kim SH, Shin HI, Bae YC, Hong JM, Park EK - PLoS ONE (2015)

Bottom Line: Abnormally elevated formation and activation of osteoclasts are primary causes for a majority of skeletal diseases.Moreover, actin ring and resorption pit formation were inhibited by KP-A159.Analysis of the signaling pathway involved showed that KP-A159 inhibited RANKL-induced activation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and mitogen-activated protein kinase kinase1/2 (MEK1/2).

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.

ABSTRACT
Abnormally elevated formation and activation of osteoclasts are primary causes for a majority of skeletal diseases. In this study, we found that KP-A159, a newly synthesized thiazolopyridine derivative, inhibited osteoclast differentiation and function in vitro, and inflammatory bone loss in vivo. KP-A159 did not cause a cytotoxic response in bone marrow macrophages (BMMs), but significantly inhibited the formation of multinucleated tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts induced by macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL). KP-A159 also dramatically inhibited the expression of marker genes related to osteoclast differentiation, including TRAP (Acp5), cathepsin K (Ctsk), dendritic cell-specific transmembrane protein (Dcstamp), matrix metallopeptidase 9 (Mmp9), and nuclear factor of activated T-cells, cytoplasmic 1 (Nfatc1). Moreover, actin ring and resorption pit formation were inhibited by KP-A159. Analysis of the signaling pathway involved showed that KP-A159 inhibited RANKL-induced activation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and mitogen-activated protein kinase kinase1/2 (MEK1/2). In a mouse inflammatory bone loss model, KP-A159 significantly rescued lipopolysaccharide (LPS)-induced bone loss by suppressing osteoclast numbers. Therefore, KP-A159 targets osteoclasts, and may be a potential candidate compound for prevention and/or treatment of inflammatory bone loss.

No MeSH data available.


Related in: MedlinePlus

Effects of KP-A159 on RANKL-induced osteoclast differentiation.(A) BMMs were cultured for 4 or 5 days in the presence of M-CSF (10 ng/mL) and RANKL (20 ng/mL) with or without 1 μM or 5 μM KP-A159. Osteoclasts were stained with TRAP. (B) TRAP-positive multinucleated cells with ≥3 nuclei were counted. ** p < 0.01 versus vehicle-treated control. (C) and (D) BMMs were cultured for 3 days with M-CSF (10 ng/mL) either with or without RANKL (20 ng/ml) in the presence or absence of 1 μM or 5 μM KP-A159. Cell viability was evaluated by the MTT assay.
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pone.0142201.g002: Effects of KP-A159 on RANKL-induced osteoclast differentiation.(A) BMMs were cultured for 4 or 5 days in the presence of M-CSF (10 ng/mL) and RANKL (20 ng/mL) with or without 1 μM or 5 μM KP-A159. Osteoclasts were stained with TRAP. (B) TRAP-positive multinucleated cells with ≥3 nuclei were counted. ** p < 0.01 versus vehicle-treated control. (C) and (D) BMMs were cultured for 3 days with M-CSF (10 ng/mL) either with or without RANKL (20 ng/ml) in the presence or absence of 1 μM or 5 μM KP-A159. Cell viability was evaluated by the MTT assay.

Mentions: To examine the effect of KP-A159 on osteoclast differentiation, we treated BMMs, stimulated with M-CSF and RANKL, with KP-A159 (1 μM or 5 μM) and analyzed the formation of osteoclast-like cells (TRAP-positive MNCs). After 4 days of culture, TRAP-positive MNCs were generated in the positive control (Fig 2A). Compared to the control, the formation of MNCs was considerably reduced by treatment with KP-A159 in a dose-dependent manner, with the number of MNCs being decreased by 62.7% at 1 μM and 85.9% at 5 μM KP-A159 (p < 0.01; Fig 2B). The inhibitory effect was not attributable to the cytotoxicity of KP-A159 because the MTT assay showed that KP-A159 (≤5 μM) did not elicit cytotoxic responses in macrophages and pre-osteoclasts (Fig 2C and 2D). These results indicate that KP-A159 dramatically suppresses the generation of osteoclast-like MNCs from BMMs without any cytotoxic effect.


Inhibitory Effects of KP-A159, a Thiazolopyridine Derivative, on Osteoclast Differentiation, Function, and Inflammatory Bone Loss via Suppression of RANKL-Induced MAP Kinase Signaling Pathway.

Ihn HJ, Lee D, Lee T, Kim SH, Shin HI, Bae YC, Hong JM, Park EK - PLoS ONE (2015)

Effects of KP-A159 on RANKL-induced osteoclast differentiation.(A) BMMs were cultured for 4 or 5 days in the presence of M-CSF (10 ng/mL) and RANKL (20 ng/mL) with or without 1 μM or 5 μM KP-A159. Osteoclasts were stained with TRAP. (B) TRAP-positive multinucleated cells with ≥3 nuclei were counted. ** p < 0.01 versus vehicle-treated control. (C) and (D) BMMs were cultured for 3 days with M-CSF (10 ng/mL) either with or without RANKL (20 ng/ml) in the presence or absence of 1 μM or 5 μM KP-A159. Cell viability was evaluated by the MTT assay.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4633183&req=5

pone.0142201.g002: Effects of KP-A159 on RANKL-induced osteoclast differentiation.(A) BMMs were cultured for 4 or 5 days in the presence of M-CSF (10 ng/mL) and RANKL (20 ng/mL) with or without 1 μM or 5 μM KP-A159. Osteoclasts were stained with TRAP. (B) TRAP-positive multinucleated cells with ≥3 nuclei were counted. ** p < 0.01 versus vehicle-treated control. (C) and (D) BMMs were cultured for 3 days with M-CSF (10 ng/mL) either with or without RANKL (20 ng/ml) in the presence or absence of 1 μM or 5 μM KP-A159. Cell viability was evaluated by the MTT assay.
Mentions: To examine the effect of KP-A159 on osteoclast differentiation, we treated BMMs, stimulated with M-CSF and RANKL, with KP-A159 (1 μM or 5 μM) and analyzed the formation of osteoclast-like cells (TRAP-positive MNCs). After 4 days of culture, TRAP-positive MNCs were generated in the positive control (Fig 2A). Compared to the control, the formation of MNCs was considerably reduced by treatment with KP-A159 in a dose-dependent manner, with the number of MNCs being decreased by 62.7% at 1 μM and 85.9% at 5 μM KP-A159 (p < 0.01; Fig 2B). The inhibitory effect was not attributable to the cytotoxicity of KP-A159 because the MTT assay showed that KP-A159 (≤5 μM) did not elicit cytotoxic responses in macrophages and pre-osteoclasts (Fig 2C and 2D). These results indicate that KP-A159 dramatically suppresses the generation of osteoclast-like MNCs from BMMs without any cytotoxic effect.

Bottom Line: Abnormally elevated formation and activation of osteoclasts are primary causes for a majority of skeletal diseases.Moreover, actin ring and resorption pit formation were inhibited by KP-A159.Analysis of the signaling pathway involved showed that KP-A159 inhibited RANKL-induced activation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and mitogen-activated protein kinase kinase1/2 (MEK1/2).

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, School of Medicine, Kyungpook National University, Daegu, Republic of Korea.

ABSTRACT
Abnormally elevated formation and activation of osteoclasts are primary causes for a majority of skeletal diseases. In this study, we found that KP-A159, a newly synthesized thiazolopyridine derivative, inhibited osteoclast differentiation and function in vitro, and inflammatory bone loss in vivo. KP-A159 did not cause a cytotoxic response in bone marrow macrophages (BMMs), but significantly inhibited the formation of multinucleated tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts induced by macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL). KP-A159 also dramatically inhibited the expression of marker genes related to osteoclast differentiation, including TRAP (Acp5), cathepsin K (Ctsk), dendritic cell-specific transmembrane protein (Dcstamp), matrix metallopeptidase 9 (Mmp9), and nuclear factor of activated T-cells, cytoplasmic 1 (Nfatc1). Moreover, actin ring and resorption pit formation were inhibited by KP-A159. Analysis of the signaling pathway involved showed that KP-A159 inhibited RANKL-induced activation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and mitogen-activated protein kinase kinase1/2 (MEK1/2). In a mouse inflammatory bone loss model, KP-A159 significantly rescued lipopolysaccharide (LPS)-induced bone loss by suppressing osteoclast numbers. Therefore, KP-A159 targets osteoclasts, and may be a potential candidate compound for prevention and/or treatment of inflammatory bone loss.

No MeSH data available.


Related in: MedlinePlus