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Article ; Online: Development of an optogenetics tool, Opto-RANK, for control of osteoclast differentiation using blue light.

Takada, Aiko / Asano, Toshifumi / Nakahama, Ken-Ichi / Ono, Takashi / Nakata, Takao / Ishii, Tomohiro

2024 Volume 14, Issue 1, Page(s) 1749

Abstract: Optogenetics enables precise regulation of intracellular signaling in target cells. However, the application of optogenetics to induce the differentiation of precursor cells and generate mature cells with specific functions has not yet been fully ... ...

Abstract	Optogenetics enables precise regulation of intracellular signaling in target cells. However, the application of optogenetics to induce the differentiation of precursor cells and generate mature cells with specific functions has not yet been fully explored. Here, we focused on osteoclasts, which play an important role in bone remodeling, to develop a novel optogenetics tool, Opto-RANK, which can manipulate intracellular signals involved in osteoclast differentiation and maturation using blue light. We engineered Opto-RANK variants, Opto-RANKc and Opto-RANKm, and generated stable cell lines through retroviral transduction. Differentiation was induced by blue light, and various assays were conducted for functional analysis. Osteoclast precursor cells expressing Opto-RANK differentiated into multinucleated giant cells on light exposure and displayed upregulation of genes normally induced in differentiated osteoclasts. Furthermore, the differentiated cells exhibited bone-resorbing activities, with the possibility of spatial control of the resorption by targeted light illumination. These results suggested that Opto-RANK cells differentiated by light possess the features of osteoclasts, both morphological and functional. Thus, Opto-RANK should be useful for detailed spatiotemporal analysis of intracellular signaling during osteoclast differentiation and the development of new therapies for various bone diseases.
MeSH term(s)	Humans ; Osteoclasts/metabolism ; Bone Resorption/metabolism ; Blue Light ; Optogenetics ; Cell Differentiation/genetics ; RANK Ligand/metabolism
Chemical Substances	RANK Ligand
Language	English
Publishing date	2024-01-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-024-52056-w
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Article: Gap junctional intercellular communication attenuates osteoclastogenesis induced by activated osteoblasts

Kawatsura, Risa / Hara, Yusuke / Akiyama, Masako / Tachikawa, Noriko / Nakahama, Ken-ichi

Biochemical and biophysical research communications. 2022 Mar. 15, v. 597

2022

Abstract: Osteoblasts participate in both bone formation through the synthesis of extracellular matrix and osteoclast differentiation through the expression of osteoclast differentiation factor. Osteoblasts communicate with each other via gap junctions (GJ), which ...

Abstract	Osteoblasts participate in both bone formation through the synthesis of extracellular matrix and osteoclast differentiation through the expression of osteoclast differentiation factor. Osteoblasts communicate with each other via gap junctions (GJ), which enable small molecules, such as cAMP, to move to adjacent cells. Therefore, we focused on the role of cAMP propagation between osteoblasts via GJ in the osteoclast-supporting activity of osteoblasts. Osteoclast-supporting activity was evaluated by a co-culture system of osteoblasts with bone marrow-derived mononuclear cells. In this system, ablation of Gja1, a gene encoding connexin 43, in osteoblasts promoted osteoclastogenesis induced by prostaglandin E₂ (PGE₂). A phosphodiesterase 4 inhibitor increased both osteoclastogenesis and the intracellular cAMP concentration ([cAMP]ᵢ₎ in osteoblasts. Individual cell analysis of [cAMP]ᵢ in osteoblasts revealed different responses of each osteoblast to PGE₂. Moreover, measurement of real-time [cAMP]ᵢ demonstrated cAMP movement from cell to cell via GJ. The inhibition of GJ resulted in the upregulation of [cAMP]ᵢ in osteoblasts stimulated by PGE2. This study suggested that GJ intercellular communication exerts protective effects against excess osteoclastogenesis via cAMP movement between osteoblasts.
Keywords	bone formation ; cell communication ; coculture ; connexins ; extracellular matrix ; genes ; osteoblasts ; osteoclasts ; prostaglandins ; research
Language	English
Dates of publication	2022-0315
Size	p. 71-76.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	205723-2
ISSN	0006-291X ; 0006-291X
ISSN (online)	0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2022.01.118
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Article ; Online: GPR110, a receptor for synaptamide, expressed in osteoclasts negatively regulates osteoclastogenesis.

Hidaka, Shiho / Mouri, Yuki / Akiyama, Masako / Miyasaka, Naoyuki / Nakahama, Ken-Ichi

Prostaglandins, leukotrienes, and essential fatty acids

2022 Volume 182, Page(s) 102457

Abstract: Bone remodeling is precisely regulated mainly by osteoblasts and osteoclasts. Although some G-protein coupled receptors (GPCRs) were reported to play roles in osteoblast function, little is known about the roles in osteoclasts. In this study, we found, ... ...

Abstract	Bone remodeling is precisely regulated mainly by osteoblasts and osteoclasts. Although some G-protein coupled receptors (GPCRs) were reported to play roles in osteoblast function, little is known about the roles in osteoclasts. In this study, we found, for the first time, that the expression of GPR110 increased during osteoclastogenesis. GPR110 belongs to adhesion GPCR and was the functional receptor of N-docosahexaenoyl ethanolamine (also called synaptamide). Synaptamide suppressed osteoclastogenesis induced by receptor activator of nuclear factor-kappa B ligand. Considering that synaptamide is the endogenous metabolite of DHA, we hypothesized that DHA may inhibit osteoclastogenesis by affecting synaptamide/GPR110 signaling. But GPR110 knockout and subsequent rescue experiments revealed a pivotal role of GPR110 in the attenuation of osteoclastogenesis by synaptamide but not by DHA. These results suggest that synaptamide/GPR110 signaling negatively regulates osteoclastogenesis. Our study suggested that ligands of GPR110, such as synaptamide, might be a useful drug for osteoporotic patients.
MeSH term(s)	Carrier Proteins/metabolism ; Cell Differentiation ; Ethanolamines ; Humans ; Osteoblasts/metabolism ; Osteoclasts ; Osteogenesis
Chemical Substances	Carrier Proteins ; Ethanolamines ; synaptamide
Language	English
Publishing date	2022-06-03
Publishing country	Scotland
Document type	Journal Article
ZDB-ID	286714-x
ISSN	1532-2823 ; 0952-3278
ISSN (online)	1532-2823
ISSN	0952-3278
DOI	10.1016/j.plefa.2022.102457
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Article ; Online: Gap junctional intercellular communication attenuates osteoclastogenesis induced by activated osteoblasts.

Kawatsura, Risa / Hara, Yusuke / Akiyama, Masako / Tachikawa, Noriko / Nakahama, Ken-Ichi

Biochemical and biophysical research communications

2022 Volume 597, Page(s) 71–76

Abstract	Osteoblasts participate in both bone formation through the synthesis of extracellular matrix and osteoclast differentiation through the expression of osteoclast differentiation factor. Osteoblasts communicate with each other via gap junctions (GJ), which enable small molecules, such as cAMP, to move to adjacent cells. Therefore, we focused on the role of cAMP propagation between osteoblasts via GJ in the osteoclast-supporting activity of osteoblasts. Osteoclast-supporting activity was evaluated by a co-culture system of osteoblasts with bone marrow-derived mononuclear cells. In this system, ablation of Gja1, a gene encoding connexin 43, in osteoblasts promoted osteoclastogenesis induced by prostaglandin E
Language	English
Publishing date	2022-01-31
Publishing country	United States
Document type	Journal Article
ZDB-ID	205723-2
ISSN	1090-2104 ; 0006-291X ; 0006-291X
ISSN (online)	1090-2104 ; 0006-291X
ISSN	0006-291X
DOI	10.1016/j.bbrc.2022.01.118
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Article ; Online: Inhibitory effects of miRNAs in astrocytes on C6 glioma progression via connexin 43.

Fukuda, Shuhei / Akiyama, Masako / Niki, Yuki / Kawatsura, Risa / Harada, Hiroyuki / Nakahama, Ken-Ichi

Molecular and cellular biochemistry

2021 Volume 476, Issue 7, Page(s) 2623–2632

Abstract: In many types of tumor cells, cell communication via gap junction is decreased or missing. Therefore, cancer cells acquire unique cytosolic environments that differ from those of normal cells. This study assessed the differences in microRNA (miRNA) ... ...

Abstract	In many types of tumor cells, cell communication via gap junction is decreased or missing. Therefore, cancer cells acquire unique cytosolic environments that differ from those of normal cells. This study assessed the differences in microRNA (miRNA) expression between cancer and normal cells. MicroRNA microarray analysis revealed five miRNAs that were highly expressed in normal astrocytes compared with that in C6 gliomas. To determine whether these miRNAs could pass through gap junctions, connexin 43 was expressed in C6 glioma cells and co-cultured with normal astrocytes. The co-culture experiment showed the possibility that miR-152-3p and miR-143-3p propagate from normal astrocytes to C6 glioma in connexin 43-dependent and -independent manners, respectively. Moreover, we established C6 glioma cells that expressed miR-152-3p or miR-143-3p. Although the proliferation of these miRNA-expressing C6 glioma cells did not differ from that of empty vectors introduced in C6 glioma cells, cell migration and invasion were significantly decreased in C6 glioma cells expressing miR-152-3p or miR-143-3p. These results suggest the possibility that miRNA produced by normal cells attenuates tumor progression through connexin 43-dependent and -independent mechanisms.
Language	English
Publishing date	2021-03-03
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	184833-1
ISSN	1573-4919 ; 0300-8177
ISSN (online)	1573-4919
ISSN	0300-8177
DOI	10.1007/s11010-021-04118-0
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Article ; Online: Cellular communications in bone homeostasis and repair.

Nakahama, Ken-Ichi

Cellular and molecular life sciences : CMLS

2010 Volume 67, Issue 23, Page(s) 4001–4009

Abstract: Cellular communication between the bone component cells osteoblasts, osteocytes and (pre-)osteoclasts is essential for bone remodeling which maintains bone integrity. As in the remodeling of other organs, cell death is a trigger for remodeling of bone. ... ...

Abstract	Cellular communication between the bone component cells osteoblasts, osteocytes and (pre-)osteoclasts is essential for bone remodeling which maintains bone integrity. As in the remodeling of other organs, cell death is a trigger for remodeling of bone. During the systematic process of bone remodeling, direct or indirect cell-cell communication is indispensable. Thus, osteoblasts induce migration and differentiation of preosteoclasts, which is followed by bone resorption (by mature multinuclear osteoclasts). After completion of bone resorption, apoptosis of mature osteoclasts and differentiation of osteoblasts are initiated. At this time, the osteoblasts do not support osteoclast differentiation but do support bone formation. Finally, osteoblasts differentiate to osteocytes in bone or to bone lining cells on bone surfaces. In this way, old bone areas are regenerated as new bone. In this review the role of cell-cell communication in bone remodeling is discussed.
MeSH term(s)	Animals ; Bone Remodeling/physiology ; Bone and Bones/cytology ; Bone and Bones/physiology ; Cell Communication/physiology ; Cell Differentiation ; Homeostasis/physiology ; Osteoblasts/cytology ; Osteoblasts/physiology ; Osteoclasts/cytology ; Osteoclasts/physiology ; Osteocytes/cytology ; Osteocytes/physiology ; Osteogenesis/physiology ; RANK Ligand/metabolism ; Regeneration/physiology ; Stem Cells/cytology ; Stem Cells/physiology
Chemical Substances	RANK Ligand
Language	English
Publishing date	2010-08-08
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-010-0479-3
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Article ; Online: Circadian expression and specific localization of synaptotagmin17 in the suprachiasmatic nucleus, the master circadian oscillator in mammals.

Fujioka, Atsuko / Nagano, Mamoru / Ikegami, Keisuke / Masumoto, Koh-Hei / Yoshikawa, Tomoko / Koinuma, Satoshi / Nakahama, Ken-Ichi / Shigeyoshi, Yasufumi

Brain research

2022 Volume 1798, Page(s) 148129

Abstract: The localization and function of synaptotagmin (syt)17 in the suprachiasmatic nucleus (SCN) of the brain, which is the master circadian oscillator, were investigated. The Syt17 mRNA-containing neurons were mainly situated in the shell region while SYT17 ... ...

Abstract	The localization and function of synaptotagmin (syt)17 in the suprachiasmatic nucleus (SCN) of the brain, which is the master circadian oscillator, were investigated. The Syt17 mRNA-containing neurons were mainly situated in the shell region while SYT17 immunoreactive cell bodies and neural fibers were detected in the core and shell of the SCN and the subparaventricular zone (SPZ). Further, electron microscopy analysis revealed SYT17 in the rough endoplasmic reticulum (rER), Golgi apparatus (G), and large and small vesicles of neurons. Syt17 mRNA expression in the SCN showed a circadian rhythm, and light exposure at night suppressed its expression. In addition, the free running period of locomotor activity rhythm was shortened in Syt17-deletion mutant mice. These findings suggest that SYT17 is involved in the regulation of circadian rhythms.
MeSH term(s)	Animals ; Mice ; Circadian Rhythm/physiology ; Mammals/genetics ; Neurons/metabolism ; RNA, Messenger/metabolism ; Suprachiasmatic Nucleus/metabolism ; Synaptotagmins/metabolism
Chemical Substances	RNA, Messenger ; Synaptotagmins (134193-27-4) ; Syt17 protein, mouse
Language	English
Publishing date	2022-11-01
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1200-2
ISSN	1872-6240 ; 0006-8993
ISSN (online)	1872-6240
ISSN	0006-8993
DOI	10.1016/j.brainres.2022.148129
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Article: Genetic and epigenetic regulation of osteopontin by cyclic adenosine 3′ 5′-monophosphate in osteoblasts

Miki, Hirohito / Okito, Asuka / Akiyama, Masako / Ono, Takashi / Tachikawa, Noriko / Nakahama, Ken-ichi

Gene. 2020 Dec. 30, v. 763

2020

Abstract: Osteopontin (OPN) is not only a marker of osteoblasts but it is also related to cancer progression and inflammation. The expression of OPN increases in response to inflammatory cytokines, hormones, and mechanical stress. Among them, cyclic-AMP (cAMP) ... ...

Abstract	Osteopontin (OPN) is not only a marker of osteoblasts but it is also related to cancer progression and inflammation. The expression of OPN increases in response to inflammatory cytokines, hormones, and mechanical stress. Among them, cyclic-AMP (cAMP) elevating agents stimulate OPN expression in the presence of 1, 25-OH vitamin D₃ (VD₃). We aimed to clarify the mechanism by which cAMP enhances OPN expression in osteoblastic cells. The OPN promoter (−2335 to +76, OPNp2335) exerted a cell type specific response to forskolin (FK) and VD₃. Sequential deletion analysis of OPNp revealed that the OPNp (−833 to +76) contained essential responsive regions to respond to cAMP signaling. In particular, both Vitamin D response element (VDRE, −758 to −743) and osteoblast-specific cis- acting element 2 (OSE2, −695 to −690) were essential for cAMP-mediated OPNp activity. The expression of vitamin D receptor (VDR), but not runt-related transcription factor 2 (Runx2), a nuclear receptor for OSE2, was induced by the treatment of the cells with FK. Although, VD₃-induced OPNp activity was slightly enhanced in VDR-overexpressing osteoblasts, it reached the same level as that of osteoblasts induced by both VD₃ and FK in the presence of histone deacetylase (HDAC) inhibitor. Moreover, we identified histone acetylation on the OPN promoter region by FK treatment. These results strongly suggest that OPNp activity is controlled by the cAMP signaling via genetic and epigenetic regulations.
Keywords	acetylation ; adenosine ; calcitriol receptors ; cyclic AMP ; cytokines ; epigenetics ; forskolin ; genes ; histone deacetylase ; histones ; hormones ; inflammation ; mechanical stress ; neoplasm progression ; osteoblasts ; osteopontin ; promoter regions ; transcription factors ; vitamin D
Language	English
Dates of publication	2020-1230
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-light
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2020.145059
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Article ; Online: Genetic and epigenetic regulation of osteopontin by cyclic adenosine 3' 5'-monophosphate in osteoblasts.

Miki, Hirohito / Okito, Asuka / Akiyama, Masako / Ono, Takashi / Tachikawa, Noriko / Nakahama, Ken-Ichi

Gene

2020 Volume 763, Page(s) 145059

Abstract	Osteopontin (OPN) is not only a marker of osteoblasts but it is also related to cancer progression and inflammation. The expression of OPN increases in response to inflammatory cytokines, hormones, and mechanical stress. Among them, cyclic-AMP (cAMP) elevating agents stimulate OPN expression in the presence of 1, 25-OH vitamin D
MeSH term(s)	Acetylation ; Animals ; Cyclic AMP/metabolism ; Epigenesis, Genetic ; HEK293 Cells ; Histone Code ; Humans ; Mice ; Osteoblasts/metabolism ; Osteopontin/chemistry ; Osteopontin/genetics ; Osteopontin/metabolism ; Promoter Regions, Genetic ; Protein Domains ; Receptors, Calcitriol/genetics ; Receptors, Calcitriol/metabolism ; Vitamin D/metabolism
Chemical Substances	Receptors, Calcitriol ; Osteopontin (106441-73-0) ; Vitamin D (1406-16-2) ; Cyclic AMP (E0399OZS9N)
Language	English
Publishing date	2020-08-25
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	391792-7
ISSN	1879-0038 ; 0378-1119
ISSN (online)	1879-0038
ISSN	0378-1119
DOI	10.1016/j.gene.2020.145059
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Article: Augmented effect of fibroblast growth factor 18 in bone morphogenetic protein 2-induced calvarial bone healing by activation of CCL2/CCR2 axis on M2 macrophage polarization.

Namangkalakul, Worachat / Nagai, Shigenori / Jin, Chengxue / Nakahama, Ken-Ichi / Yoshimoto, Yuki / Ueha, Satoshi / Akiyoshi, Kazunari / Matsushima, Kouji / Nakashima, Tomoki / Takechi, Masaki / Iseki, Sachiko

Journal of tissue engineering

2023 Volume 14, Page(s) 20417314231187960

Abstract: Fibroblast growth factor (FGF) signaling plays essential roles in various biological events. FGF18 is one of the ligands to be associated with osteogenesis, chondrogenesis and bone healing. The mouse critical-sized calvarial defect healing induced by the ...

Abstract	Fibroblast growth factor (FGF) signaling plays essential roles in various biological events. FGF18 is one of the ligands to be associated with osteogenesis, chondrogenesis and bone healing. The mouse critical-sized calvarial defect healing induced by the bone morphogenetic protein 2 (BMP2)-hydrogel is stabilized when FGF18 is added. Here, we aimed to investigate the role of FGF18 in the calvarial bone healing model. We first found that FGF18 + BMP2 hydrogel application to the calvarial bone defect increased the expression of anti-inflammatory markers, including those related to tissue healing M2 macrophage (M2-Mø) prior to mineralized bone formation. The depletion of macrophages with clodronate liposome hindered the FGF18 effect. We then examined how FGF18 induces M2-Mø polarization by using mouse primary bone marrow (BM) cells composed of macrophage precursors and BM stromal cells (BMSCs). In vitro studies demonstrated that FGF18 indirectly induces M2-Mø polarization by affecting BMSCs. Whole transcriptome analysis and neutralizing antibody treatment of BMSC cultured with FGF18 revealed that chemoattractant chemokine (c-c motif) ligand 2 (CCL2) is the major mediator for M2-Mø polarization. Finally, FGF18-augmented activity toward favorable bone healing with BMP2 was diminished in the calvarial defect in
Language	English
Publishing date	2023-07-26
Publishing country	England
Document type	Journal Article
ZDB-ID	2573915-3
ISSN	2041-7314
ISSN	2041-7314
DOI	10.1177/20417314231187960
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