title: Bone marrow metastatic myeloma cells promote osteoclastogenesis through RANKL on endothelial cells.
abstract: We have been using the B9/BM1 murine bone marrow metastasis model to study the function of adhesion molecules in the cell-cell interactions and transendothelial migration, necessary for tumor metastasis. The cell surface phenotype of these cells, which colonize vertebral and femoral marrow after intravenous injection, shows great similarity to that of human myeloma cells. In the present study, we investigated the interaction between B9/BM1 cells and osteoclasts, which likely support tumor metastasis in bone marrow. We found that co-culturing B9/BM1 cells and bone marrow-derived endothelial cells (BMECs) in the presence of vitamin D3 and M-CSF promoted differentiation of primary osteoclast progenitors to osteoclasts (detected by TRAP staining), and that this effect was blocked when BMECs were separated from the other cells by a porous polycarbonate membrane. Flow cytometry analysis showed that BMECs expressed RANKL (receptor activator of NF-kappaB ligand) protein on their surface, and that this expression was up-regulated by co-culture with B9/BM1 cells. Accordingly, RT-PCR showed expression of RANKL mRNA also to be up-regulated in BMECs co-cultured with B9/BM1 cells. Addition of OPG (osteoprotegerin, a decoy RANKL receptor) to the co-culture system completely blocked osteoclast induction, as did addition of anti-CD44 antibody. Furthermore, intravenous injection of B9/BM1 cells substantially increased the numbers of TRAP-positive osteoclasts detected in mice in vivo. Taken together, these findings suggest that B9/BM1 myeloma cells act via CD44 to stimulate RANKL expression on BMECs, which in turn physically interact with osteoclast progenitors to promote their differentiation to osteoclasts and metastasis in bone marrow.
authors.affiliation: National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan. t.okada@aist.go.jp
authors.forename: Tomoko
authors.initials: T
authors.lastname: Okada
authors.forename: Shingo
authors.initials: S
authors.lastname: Akikusa
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okuno
authors.forename: Masato
authors.initials: M
authors.lastname: Kodaka
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000502:physiology*; D019046:Bone Marrow Neoplasms / Q000556:secondary*; D002450:Cell Communication; D002454:Cell Differentiation; D018920:Coculture Techniques; D042783:Endothelial Cells / Q000502:physiology*; D005260:Female; D006023:Glycoproteins / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D051379:Mice; D009101:Multiple Myeloma / Q000556:secondary*; D010010:Osteoclasts / Q000502:physiology*; D053244:Osteoprotegerin; D018160:Receptors, Cytoplasmic and Nuclear / Q000378:metabolism*; D018124:Receptors, Tumor Necrosis Factor; D014407:Tumor Cells, Cultured
chemical_list: D006023:Glycoproteins; D015850:Interleukin-6; D053244:Osteoprotegerin; D018160:Receptors, Cytoplasmic and Nuclear; D018124:Receptors, Tumor Necrosis Factor; C506030:TNFRSF11B protein, human; C506031:Tnfrsf11b protein, mouse