Lactosylceramide Is Essential for the Osteoclastogenesis Mediated by Macrophage-Colony-stimulating Factor and Receptor Activator of Nuclear Factor-κB Ligand

Iwamoto, Tsutomu; Fukumoto, Satoshi; Kanaoka, Kazuhiro; Sakai, Eiko; Shibata, Mitsue; Fukumoto, Emiko; Inokuchi, Jin-ichi; Takamiya, Kogo; Furukawa, Koichi; Kato, Yuzo; Mizuno, Akio

doi:10.1074/jbc.m104464200

Cited by 52 publications

(39 citation statements)

References 64 publications

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“…L-Ser is known to be metabolized to ceramide/sphingolipids in mammalian cells through the condensation of serine and palmitoyl-CoA by SPT as a first step, producing KDS. In this regard, an important role for glycosphingolipid in osteoclastogenesis was reported (17). The involvement of this pathway in the formation of MN cells was actually confirmed in our system using myriocin, an SPT inhibitor.…”

Section: L-ser Metabolism Is Important For Osteoclastogenesis and Thsupporting

confidence: 62%

“…Therefore, it seems that reduced levels of sphingolipid metabolites caused by the down-regulation of SPT activity by the analog resulted in the down-regulation of RANK expression and the modulation of its localization in membrane lipid rafts. Regarding this, glycosphingolipid was reported to play an important role in osteoclastogenesis (17,21). D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol, a gycosylceramide synthase inhibitor, was shown to completely inhibit the formation of osteoclasts among bone marrow cells, and the addition of LacCer rescued the formation of TRAP-positive mononucleated cells but not multinucleated cells (17).…”

mentioning

confidence: 99%

“…Regarding this, glycosphingolipid was reported to play an important role in osteoclastogenesis (17,21). D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol, a gycosylceramide synthase inhibitor, was shown to completely inhibit the formation of osteoclasts among bone marrow cells, and the addition of LacCer rescued the formation of TRAP-positive mononucleated cells but not multinucleated cells (17). Here the addition of LacCer to the analog-treated RAW264 cells rescued the suppressed formation of TRAP-positive MN cells accompanying an up-regulation of NFAT2 expression.…”

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confidence: 99%

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Identification of a Novel l-Serine Analog That Suppresses Osteoclastogenesis in Vitro and Bone Turnover in Vivo

Bahtiar¹,

Μatsumoto²,

Nakamura

et al. 2009

Journal of Biological Chemistry

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Osteoclasts are multinucleated giant cells with bone resorbing activity. We previously reported that the expression of the transcription factor NFAT2 (NFATc1) induced by receptor activator of NF-B ligand (RANKL) is essential for the formation of multinucleated cells. We subsequently identified L-Ser in the differentiation medium as necessary for the expression of NFAT2. Here we searched for serine analogs that antagonize the function of L-Ser and suppress the formation of osteoclasts in bone marrow as well as RAW264 cells. An analog thus identified, H-Ser(tBu)-OMe⅐HCl, appeared to suppress the production of 3-ketodihydrosphingosine by serine palmitoyltransferase, and the expression and localization of RANK, a cognate receptor of RANKL, in membrane lipid rafts was down-regulated in the analog-treated cells. The addition of lactosylceramide, however, rescued the osteoclastic formation. When administered in vivo, the analog significantly increased bone density in mice and prevented high bone turnover induced by treatment with soluble RANKL. These results demonstrate a close connection between the metabolism of L-Ser and bone remodeling and also the potential of the analog as a novel therapeutic tool for bone destruction.Osteoclasts are multinucleated giant cells with bone resorbing activity and play a key role in bone remodeling in conjunction with osteoblasts (1, 2). It is well established that receptor activator of NF-B ligand (RANKL) plays a central role in osteoclastogenesis through its cognate receptor activator of NF-B (RANK) (3, 4). We previously found, using an in vitro model of osteoclastogenesis consisting of mouse cells and recombinant RANKL, that the expression of the transcription factor NFAT2 (NFATc1) induced by stimulation with RANKL is essential for the formation of mature osteoclasts (5). Interestingly, culture at high cell density in the in vitro differentiation system blocked progression to the multinucleated (MN) 3 cell stage. Subsequently, a high cell density was found to cause a change in the composition/state of the culture medium accompanying downregulation of NFAT2 expression, and we eventually identified L-Ser as a key component for the phenomenon (6). Although the differentiation medium contained seven nonessential amino acids (L-Ala, L-Asn, L-Asp, L-Glu, L-Gly, L-Pro, and L-Ser), no other amino acid showed such a characteristic property. In fact, no osteoclasts formed when only L-Ser was depleted from the differentiation medium with dialyzed serum. In this regard, D-Ser was completely inert, and moreover, when D-Ser was added together with L-Ser, there was a suppressive effect on NFAT2 expression/osteoclastic formation, implying certain analog(s) to be useful for suppressing osteoclastogenesis through the down-regulation of NFAT2 expression. Unfortunately, however, D-Ser had a toxic effect in RAW264 cells.Here we systematically searched for analogs with more effective suppressive activity and less toxicity. We consequently identified a novel serine analog, H-Ser(tBu)-OMe⅐HCl (or Ot.-Buty...

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Section: L-ser Metabolism Is Important For Osteoclastogenesis and Thsupporting

confidence: 62%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Identification of a Novel l-Serine Analog That Suppresses Osteoclastogenesis in Vitro and Bone Turnover in Vivo

Bahtiar¹,

Μatsumoto²,

Nakamura

et al. 2009

Journal of Biological Chemistry

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“…MC3T3-E1 and RAW264.7, NIH3T3, and J774 cells were maintained in ␣-MEM, Dulbecco's modified Eagle's medium, and RPMI 1640 (Invitrogen, Carlsbad, CA), respectively, containing 10% fetal bovine serum (FBS; Invitrogen) and 100 units/ml of penicillin-G at 37°C in a humidified atmosphere of 5% CO 2 in air. Bone marrow cells were obtained from tibias and femurs of 4-week-old male mice (ddY strain) and passed through a Sephadex G-10 column (Amersham Biosciences, Piscataway, NJ) as described previously (25)(26)(27) to partially enrich osteoclast precursor cells. G-10 column-eluted bone marrow cells were then cultured at 6.5 ϫ 10 5 cells per well in 96-well plates for 3 or 5 days in culture medium consisting of ␣-MEM and 10% FBS at 37°C in a humidified atmosphere of 5% CO 2 in air.…”

Section: Methodsmentioning

confidence: 99%

Infection-induced Up-regulation of the Costimulatory Molecule 4-1BB in Osteoblastic Cells and Its Inhibitory Effect on M-CSF/RANKL-induced in Vitro Osteoclastogenesis

Saito

Ohara

Hotokezaka

et al. 2004

Journal of Biological Chemistry

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Bacterial infection sometimes impairs bone metabolism. In this study, we infected the osteoblastic cell line MC3T3-E1 with Mycobacterium bovis bacillus CalmetteGué rin (BCG) and identified genes that were up-regulated in the BCG-infected cells by the suppression subtractive hybridization method. A gene encoding 4-1BB (CD137), a member of the tumor necrosis factor-␣ receptor family, was found to be one of the up-regulated genes. Up-regulation of 4-1BB was also observed by infection with Escherichia coli, Salmonella typhimurium, and Tuberculosis is an ancient disease but still ranks as one of the foremost killers of the 21st century. About one-third of the world's population is infected with Mycobacterium tuberculosis and more than two million people die annually from tuberculosis (1). Most tuberculosis cases are pulmonary, but the microorganism sometimes invades the central nervous system, the lymphatic system, and the musculoskeletal system, or is sometimes disseminated throughout the whole body. Mycobacterium infection in bones and joints presents ϳ10% of the extra-pulmonary cases (2). Vertebral osteomyelitis and osteitis also occur as a complication of vaccination with bacillus Calmette-Guérin (BCG) 1 or intravesical use of BCG (3, 4). Osteomyelitis and osteitis caused by M. tuberculosis or BCG infection sometimes lead to vertebral caries in some patients as a result of increased bone resorption.Bone is maintained by dynamic equilibrium between osteoblasts and osteoclasts in bone marrow cells. M. tuberculosis and BCG infection of bone alters this equilibrium, resulting in the loss of extracellular matrix and collapse of bone, especially vertebrae. Whether this bone resorption is attributable to direct effects of the bacteria on bone cells or infection-induced activation of inflammatory cells has not been clarified. However, a few studies have shown that mycobacterial antigens interfere with bone metabolism. Wax D, a mycobacterial cell wall peptidoglycan fragment-arabinogalactan-mycolic acid complex, induced reactive bone formation accompanied with osteomyelitis in Buffalo rats (5). Heat shock protein 10 of M. tuberculosis stimulates bone resorption in bone explanting cultures and induces osteoclast recruitment but inhibits proliferation of an osteoblast bone-forming cell line (6). The secreted protein MPB70 of BCG has significant homology with four repeat domains of osteoblast-specific factor 2/periostin (7). Epidemiological study suggests that MPB70-overproducing strains of BCG seem to be associated with an increased incidence of osteitis after BCG vaccination of neonates (7).Osteoblasts express various enzymatic markers such as alkaline phosphatase (ALP) and produce collagenous and noncol-

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“…For example, lactosylceramide (LacCer), a pivotal intermediate in the degradation and synthesis of many complex glycosphingolipids, including gangliosides, [1][2][3] is known to be involved in cell-cell and cell-matrix interactions and in signaling events linked to cell differentiation, development, apoptosis, and oncogenesis. 4,5) These applications, however, appear to have limited applicability, due to the difficulties in producing naturally occurring glycolipids such LacCer in large quantities and in purified forms. The commercial production of liposomes containing such naturally occurring substance is therefore unfeasible.…”

mentioning

confidence: 99%

Design and Facile Synthesis of Neoglycolipids as Lactosylceramide Mimetics and Their Transformation into Glycoliposomes

Harada

Murata

Totani

et al. 2005

Bioscience, Biotechnology, and Biochemistry

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Lactosylceramide Is Essential for the Osteoclastogenesis Mediated by Macrophage-Colony-stimulating Factor and Receptor Activator of Nuclear Factor-κB Ligand

Cited by 52 publications

References 64 publications

Identification of a Novel l-Serine Analog That Suppresses Osteoclastogenesis in Vitro and Bone Turnover in Vivo

Identification of a Novel l-Serine Analog That Suppresses Osteoclastogenesis in Vitro and Bone Turnover in Vivo

Infection-induced Up-regulation of the Costimulatory Molecule 4-1BB in Osteoblastic Cells and Its Inhibitory Effect on M-CSF/RANKL-induced in Vitro Osteoclastogenesis

Design and Facile Synthesis of Neoglycolipids as Lactosylceramide Mimetics and Their Transformation into Glycoliposomes

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