The induction of RANKL molecule clustering could stimulate early osteoblast differentiation
Abstract
We recently found that the membrane-bound receptor activator of NF-κB ligand (RANKL) on osteoblasts works as a receptor to stimulate osteoblast differentiation, however, the reason why the RANKL-binding molecules stimulate osteoblast differentiation has not been well clarified. Since the induction of cell-surface receptor clustering is known to lead to cell activation, we hypothesized that the induction of membrane-RANKL clustering on osteoblasts might stimulate osteoblast differentiation. Immunoblotting showed that the amount of RANKL on the membrane was increased by the RANKL-binding peptide OP3-4, but not by osteoprotegerin (OPG), the other RANKL-binding molecule, in Gfp-Rankl-transfected ST2 cells. Observation under a high-speed atomic force microscope (HS-AFM) revealed that RANKL molecules have the ability to form clusters. The induction of membrane-RANKL-OPG-Fc complex clustering by the addition of IgM in Gfp-Rankl-transfected ST2 cells could enhance the expression of early markers of osteoblast differentiation to the same extent as OP3-4, while OPG-Fc alone could not. These results suggest that the clustering-formation of membrane-RANKL on osteoblasts could stimulate early osteoblast differentiation.
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Multimeric assembly of RANKL can activate reverse signaling.
“In contrast, the multimeric assembly of RANKL can activate reverse signaling.”
From Bone remodeling: an operational process ensuring survival and bone mechanical competence (2022) · cites this paper as [246]
OP3-4, a RANKL-binding peptide that induces clustering of RANKL on the cell membrane, induces activation of RANKL reverse signaling.
“In fact, OP3-4, a RANKL-binding peptide that induces the clustering of RANKL on the cell membrane, induces the activation of RANKL reverse signaling.246”
From Bone remodeling: an operational process ensuring survival and bone mechanical competence (2022) · cites this paper as [246]
Cites
Osteoclast differentiation and activation
Boyle, W. J., Simonet, W. S., & Lacey, D. L. (2003) · Nature · doi:10.1038/nature01658 · Cited by 6,670 · see citing works
Osteoprotegerin ligand is a cytokine that regulates osteoclast differentiation and activation.
Lacey, D. L., Timms, E., Tan, H.-L., Kelley, M. J., Dunstan, C. R., Burgess, T., Elliott, R., Colombero, A., Elliott, G., Scully, S., et al. (1998) · Cell · doi:10.1016/s0092-8674(00)81569-x · Cited by 5,439 · see citing works
Osteoprotegerin: a novel secreted protein involved in the regulation of bone density.
Simonet, W. S., Lacey, D. L., Dunstan, C. R., Kelley, M., Chang, M.-S., Lüthy, R., Nguyen, H. Q., Wooden, S., Bennett, L., Boone, T., et al. (1997) · Cell · doi:10.1016/s0092-8674(00)80209-3 · Cited by 5,130 · see citing works
Functions of RANKL/RANK/OPG in bone modeling and remodeling
Boyce, B. F., & Xing, L. (2008) · Archives of Biochemistry and Biophysics · doi:10.1016/j.abb.2008.03.018 · Cited by 1,844 · see citing works
Dimerization of cell surface receptors in signal transduction
Heldin, C.-H. (1995) · Cell · doi:10.1016/0092-8674(95)90404-2 · Cited by 1,634 · see citing works
Coupling of bone resorption and formation by RANKL reverse signalling
Ikebuchi, Y., Aoki, S., Honma, M., Hayashi, M., Sugamori, Y., Khan, M., Kariya, Y., Kato, G., Tabata, Y., Penninger, J. M., et al. (2018) · Nature · doi:10.1038/s41586-018-0482-7 · Cited by 557 · see citing works
Self-citationFilming Biomolecular Processes by High-Speed Atomic Force Microscopy
Ando, T., Uchihashi, T., & Simon Scheuring, A. (2014) · Chemical Reviews · doi:10.1021/cr4003837 · Cited by 398 · see citing works
Self-citationUchihashi, T., Kodera, N., & Ando, T. (2012) · Nature Protocols · doi:10.1038/nprot.2012.047 · Cited by 289 · see citing works
Self-citationDiscovery and characterization of a selective, nonpeptidyl thrombopoietin receptor agonist
Erickson‐Miller, C. L., Delorme, E., Tian, S., Hopson, C. B., Stark, K., Giampa, L., Valoret, E. I., Duffy, K. J., Luengo, J. L., Rosen, J., et al. (2005) · Experimental Hematology · doi:10.1016/j.exphem.2004.09.006 · Cited by 208 · see citing works
RANKL Inhibition Through Osteoprotegerin Blocks Bone Loss in Experimental Periodontitis
Jin, Q., Cirelli, J. A., Park, C. H., Sugai, J. V., Taba, M., Kostenuik, P. J., & Giannobile, W. V. (2007) · Journal of Periodontology · doi:10.1902/jop.2007.070073 · Cited by 198 · see citing works
Aoki, K., Saito, H., Itzstein, C., Ishiguro, M., Shibata, T., Blanqué, R., Mian, A. H., Takahashi, M., Suzuki, Y., Yoshimatsu, M., et al. (2006) · Journal of Clinical Investigation · doi:10.1172/jci22513 · Cited by 135 · see citing works
Self-citationCheng, X., Kinosaki, M., Takami, M., Choi, Y., Zhang, H., & Murali, R. (2004) · Journal of Biological Chemistry · doi:10.1074/jbc.m309690200 · Cited by 92 · see citing works
Self-citationFuruya, Y., Inagaki, A., Khan, M., Mori, K., Penninger, J. M., Nakamura, M., Udagawa, N., Aoki, K., Ohya, K., Uchida, K., et al. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.426080 · Cited by 77 · see citing works
Self-citationSeparating T Cell Targeting Components onto Magnetically Clustered Nanoparticles Boosts Activation
Kosmides, A. K., Necochea, K., Hickey, J. W., & Schneck, J. P. (2018) · Nano Letters · doi:10.1021/acs.nanolett.7b05284 · Cited by 66 · see citing works
Saito, H., Kojima, T., Takahashi, M., Horne, W. C., Baron, R., Amagasa, T., Ohya, K., & Aoki, K. (2007) · Arthritis & Rheumatism · doi:10.1002/art.22495 · Cited by 59 · see citing works
Self-citationOminsky, M. S., Kostenuik, P. J., Cranmer, P., Smith, S. Y., & Atkinson, J. E. (2007) · Osteoporosis International · doi:10.1007/s00198-007-0363-7 · Cited by 51 · see citing works
Cytokine receptor dimerization and activation: prospects for small molecule agonists
Boger, D. L., & Goldberg, J. (2001) · Bioorganic & Medicinal Chemistry · doi:10.1016/s0968-0896(00)00276-5 · Cited by 47 · see citing works
Fei, Q., Guo, C.-J., Xu, X., Jianjun, G., Zhang, J., Chen, T., & Cui, D. (2010) · Acta Biochimica et Biophysica Sinica · doi:10.1093/abbs/gmq086 · Cited by 40 · see citing works
Vps33a Mediates RANKL Storage in Secretory Lysosomes in Osteoblastic Cells
Kariya, Y., Honma, M., Aoki, S., Chiba, A., & Suzuki, H. (2009) · Journal of Bone and Mineral Research · doi:10.1359/jbmr.090409 · Cited by 36 · see citing works
Self-citationKariya, Y., Honma, M., Hanamura, A., Aoki, S., Ninomiya, T., Nakamichi, Y., Udagawa, N., & Suzuki, H. (2010) · Journal of Bone and Mineral Research · doi:10.1002/jbmr.268 · Cited by 36 · see citing works
Self-citationSugamori, Y., Mise‐Omata, S., Maeda, C., Aoki, S., Tabata, Y., Murali, R., Yasuda, H., Udagawa, N., Suzuki, H., Honma, M., et al. (2016) · BioEssays · doi:10.1002/bies.201600104 · Cited by 31 · see citing works
Self-citationDelivery of RANKL-Binding Peptide OP3-4 Promotes BMP-2–Induced Maxillary Bone Regeneration
Uehara, T., Mise‐Omata, S., Matsui, M., Tabata, Y., Murali, R., Miyashin, M., & Aoki, K. (2016) · Journal of Dental Research · doi:10.1177/0022034516633170 · Cited by 27 · see citing works
Self-citationHigh- and low-dose OPG–Fc cause osteopetrosis-like changes in infant mice
Bargman, R., Posham, R., Boskey, A. L., Carter, E. M., DiCarlo, E. F., Verdelis, K., Raggio, C., & Pleshko, N. L. (2012) · Pediatric Research · doi:10.1038/pr.2012.118 · Cited by 26 · see citing works
Arai, Y., Aoki, K., Shimizu, Y., Tabata, Y., Ono, T., Murali, R., Mise‐Omata, S., & Wakabayashi, N. (2016) · European Journal of Pharmacology · doi:10.1016/j.ejphar.2016.04.049 · Cited by 23 · see citing works
Self-citationThe local administration of TNF-α and RANKL antagonist peptide promotes BMP-2-induced bone formation
Khan, A. A. M., Alles, N., Soysa, N. S., Mamun, M. A. A., Nagano, K., Mikami, R., Furuya, Y., Yasuda, H., Ohya, K., & Aoki, K. (2013) · Journal of Oral Biosciences · doi:10.1016/j.job.2012.12.005 · Cited by 21 · see citing works
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Bolamperti, S., Villa, I., & Rubinacci, A. (2022) · Bone Research · doi:10.1038/s41413-022-00219-8 · Cited by 482 · see citing works
Takegahara, N., Kim, H., & Choi, Y. (2022) · Bone · doi:10.1016/j.bone.2022.116353 · Cited by 129 · see citing works
The RANK/RANKL/OPG system and tumor bone metastasis: Potential mechanisms and therapeutic strategies
Zhang, Y., Liang, J., Liu, P., Wang, Q., Liu, L., & Zhao, H. (2022) · Frontiers in Endocrinology · doi:10.3389/fendo.2022.1063815 · Cited by 111 · see citing works
The RANKL-RANK Axis: A Bone to Thymus Round Trip
Sobacchi, C., Menale, C., & VILLA, A. M. (2019) · Frontiers in Immunology · doi:10.3389/fimmu.2019.00629 · Cited by 65 · see citing works
Wei, Y., Tang, C., Zhang, J., Li, Z., Zhang, X., Miron, R. J., & Zhang, Y. (2019) · Biochemical and Biophysical Research Communications · doi:10.1016/j.bbrc.2019.04.029 · Cited by 61 · see citing works
Wang, W., Zhang, L., Guo, C., & Han, J. (2020) · Nutrition & Metabolism · doi:10.1186/s12986-020-00449-9 · Cited by 62 · see citing works
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Wang, H., Xu, Q., Hu, H., Shi, C., Lin, Z., Jiang, H., Dong, H., & Guo, J. (2020) · International Journal of Nanomedicine · doi:10.2147/ijn.s268657 · Cited by 33 · see citing works
Hou, J. F., Tamura, Y., Lu, H., Takahashi, Y., Kasugai, S., Nakata, H., & Kuroda, S. (2022) · Nanomaterials · doi:10.3390/nano12111850 · Cited by 27 · see citing works
Self-citationMechanisms of RANKL delivery to the osteoclast precursor cell surface
Honma, M., Ikebuchi, Y., & Suzuki, H. (2020) · Journal of Bone and Mineral Metabolism · doi:10.1007/s00774-020-01157-3 · Cited by 23 · see citing works
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Xu, J., Yu, L., Ye, S., Ye, Z., Yang, L., & Xu, X. (2024) · Frontiers in Immunology · doi:10.3389/fimmu.2024.1254516 · Cited by 22 · see citing works
Xu, D., Gao, H., Lu, C., Tian, H., & Yu, X. (2022) · Frontiers in Endocrinology · doi:10.3389/fendo.2022.1066089 · Cited by 20 · see citing works
Freeman, C., S, M. D. A., & Priscilla, A. S. (2024) · Molecular Neurobiology · doi:10.1007/s12035-024-04227-z · Cited by 19 · see citing works
Luo, P., Fang, J., Yang, D., Yu, L., Chen, H., Jiang, C., Guo, R., Zhu, T., & Tang, S. (2022) · Bioengineering & Translational Medicine · doi:10.1002/btm2.10414 · Cited by 14 · see citing works
Rashed, F. M., Kamijo, S., Shimizu, Y., Hirohashi, Y., Khan, M., Sugamori, Y., Murali, R., & Aoki, K. (2021) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2021.648084 · Cited by 14 · see citing works
Self-citationRuan, S., Rody, W. J., Patel, S. S., Hammadi, L. I., Martin, M. L., de Faria, L. P., Daaboul, G. G., Anderson, L., He, M., & Holliday, L. S. (2023) · Extracellular Vesicles and Circulating Nucleic Acids · doi:10.20517/evcna.2023.38 · Cited by 7 · see citing works
Keo, P., Matsumoto, Y., Shimizu, Y., Nagahiro, S., Ikeda, M., Aoki, K., & Ono, T. (2020) · European Journal of Orthodontics · doi:10.1093/ejo/cjaa018 · Cited by 4 · see citing works
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Li, C., Yang, P., Kou, Y., Zhang, D., & Li, M. (2021) · Archives of Oral Biology · doi:10.1016/j.archoralbio.2021.105243 · Cited by 3 · see citing works
Nagahiro, S., Uehara, T., Kawai, M. Y., Keo, P., Sato, T., Ochi, H., Sato, S., Kuroda, S., Ono, T., Miyashin, M., et al. (2020) · Dental Oral and Maxillofacial Research · doi:10.15761/domr.1000327
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Rashed, F., & Aoki, K. (2025) · Journal of Sciences and Innovation for Sustainable Peace · doi:10.23184/jhja.vol.3.no.2.06
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- Authors
- Sone, E., Noshiro, D., Ikebuchi, Y., Nakagawa, M., Khan, M., Tamura, Y., Ikeda, M., Oki, M., Murali, R., Fujimori, T., Yoda, T., Honma, M., Suzuki, H., Ando, T., & Aoki, K.
- Affiliations
- Tokyo Medical and Dental University (JP)
- Kanazawa University (JP)
- University of Tokyo Hospital (JP)
- National Institute for Basic Biology (JP)
- Cedars-Sinai Medical Center (US)
- Corresponding author
- Aoki, Kazuhiro
- ORCID
- Venue
- Biochemical and Biophysical Research Communications
- Publisher
- Elsevier BV
- Published
- 2019-02
- Volume
- 509
- Issue
- 2
- Pages
- 435-440
- ISSN
- 0006-291X (ISSN-L)
- OpenAlex
- W2906550673
- Cited by
- 24
- References
- 25
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 3 · 2023: 1 · 2022: 7 · 2021: 2 · 2020: 5 · 2019: 3
- FWCI
- 0.89
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- Locations
- Biochemical and Biophysical Research Communications (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Japan Society for the Promotion of Science (16K15656, 17F17124, JP16H06280)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bone Metabolism and Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Health Sciences › Medicine › Oncology › Cytokine Signaling Pathways and Interactions
- Keywords
- RANKL, receptor clustering, osteogenic differentiation, osteoprotegerin, IgM, ST2 cells, membrane receptor signaling, high-speed atomic force microscopy
- MeSH terms
- Animals, Binding Sites, Cell Differentiation, Cell Line, Cell Membrane, Gene Expression Regulation, Immunoglobulin M, Immunoglobulin Fc Fragments, Models, Molecular, Oligopeptides, Osteoblasts, Protein Binding, Recombinant Fusion Proteins, Time Factors, Signal Transduction, Microscopy, Atomic Force, Green Fluorescent Proteins, Mice, Osteoprotegerin, RANK Ligand, Peptidomimetics
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