Cadherin-6 Mediates the Heterotypic Interactions between the Hemopoietic Osteoclast Cell Lineage and Stromal Cells in a Murine Model of Osteoclast Differentiation

Osteoclasts are multinucleated cells of hemopoietic origin that are responsible for bone resorption during physiological bone remodeling and in a variety of bone diseases. Osteoclast development requires direct heterotypic cell–cell interactions of the hemopoietic osteoclast precursors with the neighboring osteoblast/stromal cells. However, the molecular mechanisms underlying these heterotypic interactions are poorly understood. We isolated cadherin-6 isoform, denoted cadherin-6/2 from a cDNA library of human osteoclast-like cells. The isolated cadherin-6/2 is 3,423 bp in size consisting of an open reading frame of 2,115 bp, which encodes 705 amino acids. This isoform lacks 85 amino acids between positions 333 and 418 and contains 9 different amino acids in the extracellular domain compared with the previously described cadherin-6. The human osteoclast-like cells also expressed another isoform denoted cadherin-6/1 together with the cadherin-6. Introduction of cadherin-6/2 into L-cells that showed no cell–cell contact caused evident morphological changes accompanied with tight cell–cell association, indicating the cadherin-6/2 we isolated here is functional. Moreover, expression of dominant-negative or antisense cadherin-6/2 construct in bone marrow–derived mouse stromal ST2 cells, which express only cadherin-6/2, markedly impaired their ability to support osteoclast formation in a mouse coculture model of osteoclastogenesis. Our results suggest that cadherin-6 may be a contributory molecule to the heterotypic interactions between the hemopoietic osteoclast cell lineage and osteoblast/bone marrow stromal cells required for the osteoclast differentiation. Since both osteoclasts and osteoblasts/bone marrow stromal cells are the primary cells controlling physiological bone remodeling, expression of cadherin-6 isoforms in these two cell types of different origin suggests a critical role of these molecules in the relationship of osteoclast precursors and cells of osteoblastic lineage within the bone microenvironment.

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Role of neural-cadherin in early osteoblastic differentiation of human bone marrow stromal cells cocultured with human umbilical vein endothelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00562.2009 ◽

2010 ◽

Vol 299 (2) ◽

pp. C422-C430 ◽

Cited By ~ 40

Author(s):

Haiyan Li ◽

Richard Daculsi ◽

Maritie Grellier ◽

Reine Bareille ◽

Chantal Bourget ◽

...

Keyword(s):

Bone Marrow ◽

Endothelial Cells ◽

Cell Adhesion ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Umbilical Vein ◽

Osteoblastic Differentiation ◽

Marrow Stromal Cells ◽

Umbilical Vein Endothelial Cells ◽

Cell Cell

In our previous studies, roles of gap junction and vascular endothelial growth factor in the cross-talking of human bone marrow stromal cells (HBMSCs) and human umbilical vein endothelial cells (HUVECs) have been extensively studied. The present study focused on the investigation of the roles of neural (N)-cadherin in early differentiation of HBMSCs in direct-contact cocultures with HUVECs for 24 and 48 h. Quantitative real-time polymerase chain reaction, immunofluorescence, Western blot, as well as functional studies were applied to perform the studies at both protein and gene levels. Results showed that cocultured cells expressed much higher N-cadherin than monocultured cells after 24 and 48 h of culture. We observed that N-cadherin concentrated in the membrane of cocultured HBMSCs (co-HBMSCs) while distributed within the cytoplasm of monocultured HBMSCs, which indicated that the cell-cell adhesion was improved between cocultured cells. In addition, more β-catenin was found to translocate into the cocultured cells nuclei and more T cell factor-1 (TCF-1) were detected in cocultured cells than in the monocultured cells. Moreover, mRNA levels of early osteoblastic markers including alkaline phosphatase (ALP) and type I collagen (Col-I) of co-HBMSCs were significantly upregulated, whereas neutralization of N-cadherin led to a downregulation of ALP and Col-I in both of the HBMSCs and co-HBMSCs compared with untreated cells. Taking our findings together it can be concluded that cocultures of HBMSCs with HUVECs increased N-cadherin expression and improved cell-cell adhesion. Whether this applies only to osteoprogenitor cells or to all the cell types in the culture will need to be determined by further studies. Subsequently, signaling transduction might be induced with the participation of β-catenin and TCF-1. With the N-cadherin-mediated cell-cell adhesion and signaling transductions, the early osteoblastic differentiation of co-HBMSCs was significantly upregulated.

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Gefitinib reduces the production of osteoclast differentiation factors in human bone marrow stromal cells: Implications for treatment of bone metastasis

Journal of Clinical Oncology ◽

10.1200/jco.2005.23.16_suppl.3119 ◽

2005 ◽

Vol 23 (16_suppl) ◽

pp. 3119-3119

Author(s):

N. Normanno ◽

A. de Luca ◽

D. Aldinucci ◽

M. R. Maiello ◽

M. Mancino ◽

...

Keyword(s):

Bone Marrow ◽

Bone Metastasis ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Osteoclast Differentiation ◽

Marrow Stromal Cells ◽

Human Bone ◽

Human Bone Marrow ◽

Differentiation Factors

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Biglycan (Bgn) and fibromodulin (Fmod) modulate bone mass by regulating osteoclast differentiation through bone marrow stromal cells

Bone ◽

10.1016/j.bone.2008.08.058 ◽

2008 ◽

Vol 43 ◽

pp. S58

Author(s):

Yanming Bi ◽

Tina Kilts ◽

Alfred Griffin ◽

Theresa Hefferan ◽

Fatima Syed-Picard ◽

...

Keyword(s):

Bone Marrow ◽

Bone Mass ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Osteoclast Differentiation ◽

Marrow Stromal Cells

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Connexin-43-type gap junctions mediate communication between bone marrow stromal cells

Blood ◽

10.1182/blood.v82.1.38.bloodjournal82138 ◽

1993 ◽

Vol 82 (1) ◽

pp. 38-45 ◽

Cited By ~ 58

Author(s):

K Dorshkind ◽

L Green ◽

A Godwin ◽

WH Fletcher

Keyword(s):

Bone Marrow ◽

Gap Junctions ◽

Cell Line ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Stromal Cell ◽

Cell Communication ◽

Marrow Stromal Cells ◽

Stromal Cell Line ◽

Cell Cell

Several morphologic studies have suggested that gap junctions exist between bone marrow stromal cells. This possibility was examined by analysis of stromal cells present in the adherent layer of primary long- term lymphoid bone marrow cultures and in additional studies using a stromal cell line. Results showing that the fluorescent dye lucifer yellow, when microinjected into a single stromal cell, transferred between most other contacting stroma and that stromal cells were electronically coupled provided support that cell-cell communication occurs between these microenvironmental elements. Additional studies showed that transcripts for connexin (Cx) 43, but not for Cx26 or Cx32, were present in a stromal cell line. To examine the potential for regulated cell-cell communication between the stroma, cells were treated with interleukin-1 (IL-1), a cytokine known to affect stromal cell function, and the effects on dye transfer were examined. IL-1 treatment resulted in a reversible decrease in the ability of dye to transfer between stromal cells in contact. Taken together, these studies show that gap junctions exist between stromal cells and that their permeability can be regulated. However, gap junction-mediated cell-cell communication could not be shown between the stroma and developing lymphoid cells.

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Bone Marrow Stromal Cells-Induced Drug Resistance in Multiple Myeloma

International Journal of Molecular Sciences ◽

10.3390/ijms21020613 ◽

2020 ◽

Vol 21 (2) ◽

pp. 613 ◽

Cited By ~ 4

Author(s):

Roberto Ria ◽

Angelo Vacca

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Drug Resistance ◽

Plasma Cell ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Plasma Cells ◽

Complex Structure ◽

Cell Lineage ◽

Marrow Stromal Cells

Multiple myeloma is a B-cell lineage cancer in which neoplastic plasma cells expand in the bone marrow and pathophysiological interactions with components of microenvironment influence many biological aspects of the malignant phenotype, including apoptosis, survival, proliferation, and invasion. Despite the therapeutic progress achieved in the last two decades with the introduction of a more effective and safe new class of drugs (i.e., immunomodulators, proteasome inhibitors, monoclonal antibodies), there is improvement in patient survival, and multiple myeloma (MM) remains a non-curable disease. The bone marrow microenvironment is a complex structure composed of cells, extracellular matrix (ECM) proteins, and cytokines, in which tumor plasma cells home and expand. The role of the bone marrow (BM) microenvironment is fundamental during MM disease progression because modification induced by tumor plasma cells is crucial for composing a “permissive” environment that supports MM plasma cells proliferation, migration, survival, and drug resistance. The “activated phenotype” of the microenvironment of multiple myeloma is functional to plasma cell proliferation and spreading and to plasma cell drug resistance. Plasma cell drug resistance induced by bone marrow stromal cells is mediated by stress-managing pathways, autophagy, transcriptional rewiring, and non-coding RNAs dysregulation. These processes represent novel targets for the ever-increasing anti-MM therapeutic armamentarium.

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The HGF/Met/NF-κB Pathway Regulates RANKL Expression in Osteoblasts and Bone Marrow Stromal Cells

International Journal of Molecular Sciences ◽

10.3390/ijms21217905 ◽

2020 ◽

Vol 21 (21) ◽

pp. 7905

Author(s):

Masanobu Tsubaki ◽

Shiori Seki ◽

Tomoya Takeda ◽

Akiko Chihara ◽

Yuuko Arai ◽

...

Keyword(s):

Bone Marrow ◽

Bone Disease ◽

Stromal Cells ◽

Bone Marrow Stromal Cells ◽

Osteoclast Differentiation ◽

Nuclear Factor Κb ◽

Marrow Stromal Cells ◽

Nuclear Factor ◽

Rankl Expression ◽

Enhanced Expression

Multiple myeloma (MM)-induced bone disease occurs through hyperactivation of osteoclasts by several factors secreted by MM cells. MM cell-secreted factors induce osteoclast differentiation and activation via direct and indirect actions including enhanced expression of receptor activator of nuclear factor κB ligand (RANKL) in osteoblasts and bone marrow stromal cells (BMSCs). Hepatocyte growth factor (HGF) is elevated in MM patients and is associated with MM-induced bone disease, although the mechanism by which HGF promotes bone disease remains unclear. In the present study, we demonstrated that HGF induces RANKL expression in osteoblasts and BMSCs, and investigated the mechanism of induction. We found that HGF and MM cell supernatants induced RANKL expression in ST2 cells, MC3T3-E1 cells, and mouse BMSCs. In addition, HGF increased phosphorylation of Met and nuclear factor κB (NF-κB) in ST2 cells, MC3T3-E1 cells, or mouse BMSCs. Moreover, Met and NF-κB inhibitors suppressed HGF-induced RANKL expression in ST2 cells, MC3T3-E1 cells, and mouse BMSCs. These results indicated that HGF promotes RANKL expression in osteoblasts and BMSCs via the Met/NF-κB signaling pathway, and Met and NF-κB inhibitors suppressed HGF-induced RANKL expression. Our findings suggest that Met and NF-κB inhibitors are potentially useful in mitigating MM-induced bone disease in patients expressing high levels of HGF.

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