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The vitamin D analogue ED71 but Not 1,25(OH)2D3 targets HIF1α protein in osteoclasts.

Sato Y, Miyauchi Y, Yoshida S, Morita M, Kobayashi T, Kanagawa H, Katsuyama E, Fujie A, Hao W, Tando T, Watanabe R, Miyamoto K, Morioka H, Matsumoto M, Toyama Y, Miyamoto T - PLoS ONE (2014)

Bottom Line: We found that 1,25(OH)2D3 or ED71 function in osteoclasts requires the vitamin D receptor (VDR).ED71 was significantly less effective in inhibiting M-CSF and RANKL-stimulated osteoclastogenesis than was 1,25(OH)2D3 in vitro.Downregulation of c-Fos protein and induction of Ifnβ mRNA in osteoclasts, both of which reportedly block osteoclastogenesis induced by 1,25(OH)2D3 in vitro, were both significantly higher following treatment with 1,25(OH)2D3 than with ED71.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Department of Musculoskeletal Reconstruction and Regeneration Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.

ABSTRACT
Although both an active form of the vitamin D metabolite, 1,25(OH)2D3, and the vitamin D analogue, ED71 have been used to treat osteoporosis, anti-bone resorbing activity is reportedly seen only in ED71- but not in 1,25(OH)2D3 -treated patients. In addition, how ED71 inhibits osteoclast activity in patients has not been fully characterized. Recently, HIF1α expression in osteoclasts was demonstrated to be required for development of post-menopausal osteoporosis. Here we show that ED71 but not 1,25(OH)2D3, suppress HIF1α protein expression in osteoclasts in vitro. We found that 1,25(OH)2D3 or ED71 function in osteoclasts requires the vitamin D receptor (VDR). ED71 was significantly less effective in inhibiting M-CSF and RANKL-stimulated osteoclastogenesis than was 1,25(OH)2D3 in vitro. Downregulation of c-Fos protein and induction of Ifnβ mRNA in osteoclasts, both of which reportedly block osteoclastogenesis induced by 1,25(OH)2D3 in vitro, were both significantly higher following treatment with 1,25(OH)2D3 than with ED71. Thus, suppression of HIF1α protein activity in osteoclasts in vitro, which is more efficiently achieved by ED71 rather than by 1,25(OH)2D3, could be a reliable read-out in either developing or screening reagents targeting osteoporosis.

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Related in: MedlinePlus

ED71 or 1,25(OH)2D3 induce Ifnβ and suppress c-Fos protein through the VDR.(A and B) M-CSF-dependent osteoclast progenitor cells were isolated from wild-type or VDR-deficient mice and cultured in the presence of M-CSF alone (50 ng/ml) or M−CSF + RANKL (25 ng/ml) with or without 10−7 M of ED71 or 1,25(OH)2D3 (1,25D) for 5 days. Ifnβ expression was then analyzed by realtime PCR (A). Data represent mean Ifnβ expression relative to that of Actb ± SD (n = 5). c-Fos protein was analyzed by western blot (B).
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pone-0111845-g004: ED71 or 1,25(OH)2D3 induce Ifnβ and suppress c-Fos protein through the VDR.(A and B) M-CSF-dependent osteoclast progenitor cells were isolated from wild-type or VDR-deficient mice and cultured in the presence of M-CSF alone (50 ng/ml) or M−CSF + RANKL (25 ng/ml) with or without 10−7 M of ED71 or 1,25(OH)2D3 (1,25D) for 5 days. Ifnβ expression was then analyzed by realtime PCR (A). Data represent mean Ifnβ expression relative to that of Actb ± SD (n = 5). c-Fos protein was analyzed by western blot (B).

Mentions: Moreover, decreased c-Fos protein and elevated Ifnβ expression seen following treatment with 1,25(OH)2D3 or ED71 were abrogated in VDR-deficient osteoclasts (Fig. 4A and B), supporting the idea that both compounds act on osteoclasts via the VDR.


The vitamin D analogue ED71 but Not 1,25(OH)2D3 targets HIF1α protein in osteoclasts.

Sato Y, Miyauchi Y, Yoshida S, Morita M, Kobayashi T, Kanagawa H, Katsuyama E, Fujie A, Hao W, Tando T, Watanabe R, Miyamoto K, Morioka H, Matsumoto M, Toyama Y, Miyamoto T - PLoS ONE (2014)

ED71 or 1,25(OH)2D3 induce Ifnβ and suppress c-Fos protein through the VDR.(A and B) M-CSF-dependent osteoclast progenitor cells were isolated from wild-type or VDR-deficient mice and cultured in the presence of M-CSF alone (50 ng/ml) or M−CSF + RANKL (25 ng/ml) with or without 10−7 M of ED71 or 1,25(OH)2D3 (1,25D) for 5 days. Ifnβ expression was then analyzed by realtime PCR (A). Data represent mean Ifnβ expression relative to that of Actb ± SD (n = 5). c-Fos protein was analyzed by western blot (B).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4222951&req=5

pone-0111845-g004: ED71 or 1,25(OH)2D3 induce Ifnβ and suppress c-Fos protein through the VDR.(A and B) M-CSF-dependent osteoclast progenitor cells were isolated from wild-type or VDR-deficient mice and cultured in the presence of M-CSF alone (50 ng/ml) or M−CSF + RANKL (25 ng/ml) with or without 10−7 M of ED71 or 1,25(OH)2D3 (1,25D) for 5 days. Ifnβ expression was then analyzed by realtime PCR (A). Data represent mean Ifnβ expression relative to that of Actb ± SD (n = 5). c-Fos protein was analyzed by western blot (B).
Mentions: Moreover, decreased c-Fos protein and elevated Ifnβ expression seen following treatment with 1,25(OH)2D3 or ED71 were abrogated in VDR-deficient osteoclasts (Fig. 4A and B), supporting the idea that both compounds act on osteoclasts via the VDR.

Bottom Line: We found that 1,25(OH)2D3 or ED71 function in osteoclasts requires the vitamin D receptor (VDR).ED71 was significantly less effective in inhibiting M-CSF and RANKL-stimulated osteoclastogenesis than was 1,25(OH)2D3 in vitro.Downregulation of c-Fos protein and induction of Ifnβ mRNA in osteoclasts, both of which reportedly block osteoclastogenesis induced by 1,25(OH)2D3 in vitro, were both significantly higher following treatment with 1,25(OH)2D3 than with ED71.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedic Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan; Department of Musculoskeletal Reconstruction and Regeneration Surgery, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan.

ABSTRACT
Although both an active form of the vitamin D metabolite, 1,25(OH)2D3, and the vitamin D analogue, ED71 have been used to treat osteoporosis, anti-bone resorbing activity is reportedly seen only in ED71- but not in 1,25(OH)2D3 -treated patients. In addition, how ED71 inhibits osteoclast activity in patients has not been fully characterized. Recently, HIF1α expression in osteoclasts was demonstrated to be required for development of post-menopausal osteoporosis. Here we show that ED71 but not 1,25(OH)2D3, suppress HIF1α protein expression in osteoclasts in vitro. We found that 1,25(OH)2D3 or ED71 function in osteoclasts requires the vitamin D receptor (VDR). ED71 was significantly less effective in inhibiting M-CSF and RANKL-stimulated osteoclastogenesis than was 1,25(OH)2D3 in vitro. Downregulation of c-Fos protein and induction of Ifnβ mRNA in osteoclasts, both of which reportedly block osteoclastogenesis induced by 1,25(OH)2D3 in vitro, were both significantly higher following treatment with 1,25(OH)2D3 than with ED71. Thus, suppression of HIF1α protein activity in osteoclasts in vitro, which is more efficiently achieved by ED71 rather than by 1,25(OH)2D3, could be a reliable read-out in either developing or screening reagents targeting osteoporosis.

Show MeSH
Related in: MedlinePlus