Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression

Shevde, Nirupama K.; Bendixen, Amy C.; Dienger, Krista; Pike, J. Wesley

doi:10.1073/pnas.130200197

Cited by 442 publications

(324 citation statements)

References 46 publications

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“…RAW 264.7 cells have been used also to generate OCLs by other investigators. (42) Our studies complement those obtained by them, because we further characterize the cells for expression of a panel of osteoclast-specific genes. It is worth noting that although Shevde et al use M-CSF and RANKL to induce the cells to differentiate, we obtain similar results by using RANKL alone.…”

Section: Discussionsupporting

confidence: 57%

c-myc Is Required for Osteoclast Differentiation

Battaglino¹,

D²,

Fu³

et al. 2002

Journal of Bone and Mineral Research

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The role of the receptor activator of nuclear factor B (NF-B) ligand (RANKL)-a tumor necrosis factor (TNF)-related cytokine-in osteoclast formation has been established clearly. However, the downstream signaling pathways activated by this cytokine remain largely unknown. To identify genes that play a role in osteoclastogenesis, we used RAW 264.7 mouse monocytes as a model system for the differentiation of multinucleated osteoclasts from mononucleated precursors. RAW 264.7 cells were induced with RANKL to form multinucleated giant osteoclast-like cells (OCLs) that expressed a number of osteoclast-specific markers and were able to form resorption pits on both calcium phosphate films and bone slices. This system was used to identify genes that are regulated by RANKL and may play a role in osteoclast differentiation.

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Section: Discussionsupporting

confidence: 57%

c-myc Is Required for Osteoclast Differentiation

Battaglino¹,

D²,

Fu³

et al. 2002

Journal of Bone and Mineral Research

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“…Confluent cultures were treated with 1,25(OH) 2D3 or 2MD at the concentrations indicated on days 0, 3, and 6 followed by treatment with ascorbic acid (50 g͞ml) and ␤Ϫglycerophosphate (10 mM) on days 9 and 12. Cells were stained on day 14 by using the Von Kossa staining technique to detect the presence of calcified matrix͞bone (11). The dark brown to black stain is indicative of calcified bone nodules.…”

Section: Discussionmentioning

confidence: 99%

“…Primary human osteoblasts were isolated as described (11) and cultured in duplicate in 6-well plates at a density of 1 ϫ 10 5 cells͞ml in DMEM͞F12 medium with 10% FBS. Primary fetal mouse calvarial cells were isolated as described (12) and cultured in ␣MEM containing 10% FBS.…”

Section: Bone Calcium Mobilization and Intestinal Calcium Transport Smentioning

confidence: 99%

A potent analog of 1α,25-dihydroxyvitamin D ₃ selectively induces bone formation

Shevde

Plum

Clagett‐Dame

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

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171

160

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1,25-Dihydroxyvitamin D3 [1,25(OH)2D3] is a principal regulator of calcium and phosphorus homeostasis through actions on intestine, kidney, and bone. 1,25(OH) 2D3 is not considered to play a significant role in bone formation, except for its role in supporting mineralization. We report here on the properties of 2-methylene-19-nor-(20S)-1␣,25(OH) 2D3 (2MD), a highly potent analog of 1,25(OH)2D3 that induces bone formation both in vitro and in vivo. Selectivity for bone was first demonstrated through the observation that 2MD is at least 30-fold more effective than 1,25(OH) 2D3 in stimulating osteoblast-mediated bone calcium mobilization while being only slightly more potent in supporting intestinal calcium transport. 2MD is also highly potent in promoting osteoblast-mediated osteoclast formation in vitro, a process essential to both bone resorption and formation. Most significantly, 2MD at concentrations as low as 10 ؊12 M causes primary cultures of osteoblasts to produce bone in vitro. This effect is not found with 1,25(OH) 2D3 even at 10 ؊8 M, suggesting that 2MD might be osteogenic in vivo. Indeed, 2MD (7 pmol͞day) causes a substantial increase (9%) in total body bone mass in ovariectomized rats over a 23-week period. 1,25(OH) 2D3 (500 pmol three times a week) only prevented the bone loss associated with ovariectomy and did not increase bone mass. These results indicate that 2MD is a potent bone-selective analog of 1,25(OH) 2D3 potentially effective in treating bone loss diseases.V itamin D-deficiency diseases such as rickets and osteomalacia represent a failure in bone formation primarily caused by a failure of mineralization (1-3). Indeed, early investigations into the underlying defect associated with rickets revealed that incubation of bone slices in media containing physiologic levels of calcium and phosphorus resulted in normal mineralization of rachitic matrix (3). Underwood and DeLuca (4) subsequently demonstrated that the mineralization defect associated with vitamin D deficiency is due directly to insufficient calcium and phosphorus in plasma at sites of mineralization. These early conclusions have been supported independently by recent studies that show that skeletal disease, which arises from deletion of the vitamin D receptor (VDR), the exclusive transcriptional mediator of vitamin D action in vivo, can be normalized in mice through lactose-containing diets high in calcium and phosphorus (5). Thus, there is little evidence that vitamin D plays a direct role in new bone formation apart from its action to maintain calcium and phosphorus levels in the blood.We recently discovered a class of vitamin D compounds modified at the 2-carbon position of the A ring that is highly potent. The introduction of a methylene group at carbon 2, removal of a methylene group at carbon 10, and alteration of the stereochemistry of the 20-carbon to yield the 20S-derivative resulted in 2-methylene-19-nor-(20S)-1␣,25(OH) 2 D 3 (2MD), a compound that exhibits an affinity for the VDR equal to that of 1,25-dihydroxyvitamin D...

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“…Cells were cultured in phenol red-free ␣-MEM with 10% charcoal dextran-treated FBS in the presence or absence of indicated compounds and the medium replaced on day 4 and day 7. Cells were fixed and stained for tartrate-resistant acid phosphatase (TRAP) activity on day 8 -10 as previously described (24). Osteoclast number was determined by counting the total number of multinucleated (Ͼ3 nuclei), TRAP-positive cells/well.…”

Section: Reagentsmentioning

confidence: 99%

“…Serum also contains DBP, an avid binder of and presumed primary although not exclusive carrier in the blood for most vitamin D metabolites (24). In contrast to 1,25(OH) 2 D 3 , 2MD binds only weakly to DBP, an effect that could lead to an increase in its functional concentration relative to 1,25(OH) 2 D 3 .…”

Section: MD Is a 125(oh) 2 D 3 -Like Superagonist In Cell Culture-mentioning

confidence: 99%

2-Methylene-19-nor-(20S)-1,25-dihydroxyvitamin D3 Potently Stimulates Gene-specific DNA Binding of the Vitamin D Receptor in Osteoblasts

Yamamoto¹,

Shevde

Warrier

et al. 2003

Journal of Biological Chemistry

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2-Methylene-19-nor-(20S)-1,25-dihydroxyvitaminD 3 (2MD) is a highly potent analog of 1,25-dihydroxyvitamin D 3 (1,25(OH) 2 D 3 ) whose actions are mediated through the vitamin D receptor (VDR). In this report, we have replicated this increased potency of 2MD in vitro using osteoblastic cells and explored its underlying molecular mechanism. 2MD stimulates the expression of several vitamin D-sensitive genes including 25-hydroxyvitamin D 3 -24 hydroxylase (Cyp24), osteopontin and receptor activator of NFB ligand and suppresses osteoprotegerin at concentrations two logs lower than that for 1,25(OH) 2 D 3 . 2MD is also more potent in stimulating transfected chimeric reporter genes under either Cyp24 or the osteocalcin promoter control. Enhanced potency is retained regardless of medium serum content. Interestingly, the uptake of both 1,25(OH) 2 D 3 and 2MD into cells is similar, as is their rapid association with the VDR. This indicates that comparable levels of occupied VDR do not elicit equivalent levels of transactivation. Using chromatin immunoprecipitation (ChIP), however, we observed a strong correlation between DNA-bound receptor and the level of induced transcription suggesting a 2MD-induced increase in affinity of the VDR for DNA. Additional studies using a mammalian two-hybrid system and ChIP indicate that 2MD is also more potent in promoting interaction with RXR and the coactivators SRC-1 and DRIP205. Finally, protease digestion studies revealed a unique VDR conformation in the presence of 2MD. These studies suggest that the molecular mechanism of 2MD potency is due to its ability to promote enhanced levels of specific DNA binding by the VDR and could suggest possible explanations for the tissue-and gene-selective actions of 2MD.

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Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression

Cited by 442 publications

References 46 publications

c-myc Is Required for Osteoclast Differentiation

c-myc Is Required for Osteoclast Differentiation

A potent analog of 1α,25-dihydroxyvitamin D ₃ selectively induces bone formation

2-Methylene-19-nor-(20S)-1,25-dihydroxyvitamin D3 Potently Stimulates Gene-specific DNA Binding of the Vitamin D Receptor in Osteoblasts

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Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression

Cited by 442 publications

References 46 publications

c-myc Is Required for Osteoclast Differentiation

c-myc Is Required for Osteoclast Differentiation

A potent analog of 1α,25-dihydroxyvitamin D 3 selectively induces bone formation

2-Methylene-19-nor-(20S)-1,25-dihydroxyvitamin D3 Potently Stimulates Gene-specific DNA Binding of the Vitamin D Receptor in Osteoblasts

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A potent analog of 1α,25-dihydroxyvitamin D ₃ selectively induces bone formation