title: Differentiation signalobody: Demonstration of antigen-dependent osteoclast differentiation from a progenitor cell line.
abstract: A "cytokine-less" in vitro differentiation method would be promising for cost-effective mass production of cells used for regenerative medicine. In this study, we developed a differentiation signalobody S-RANK, in which the extracellular domain of receptor activator of nuclear factor kappa-B (RANK) is replaced with a single-chain variable fragment (scFv) to attain signaling in response to an inexpensive antigen. A murine macrophage cell line RAW264, which is known to differentiate into an osteoclast by RANK ligand (RANKL), was lentivirally transduced with S-RANK. When the resultant cells were cultured with a specific antigen, the cells differentiated into multinucleated tartrate-resistant acid phosphatase-positive osteoclasts. The differentiation efficiency was almost comparable to those induced by RANKL. In addition, the signaling analysis demonstrated that nuclear factor kappa-B and mitogen-activated protein kinase signaling pathways, which are the major signaling pathways downstream of wild-type RANK, were also activated by S-RANK. These results demonstrate that S-RANK sufficiently mimics signal transduction of wild-type RANK. Differentiation signalobodies may be applied for controlling differentiation of other cell types by using appropriate signaling domains.
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Hideto
authors.initials: H
authors.lastname: Nakabayashi
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Saeko
authors.initials: S
authors.lastname: Aoyama
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. Electronic address: kawahara@bio.t.u-tokyo.ac.jp.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kawahara
authors.affiliation: Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
authors.forename: Teruyuki
authors.initials: T
authors.lastname: Nagamune
mesh_terms: D000818:Animals; D000941:Antigens / Q000276:immunology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D016328:NF-kappa B / Q000819:agonists / Q000378:metabolism; D010010:Osteoclasts / Q000166:cytology* / Q000187:drug effects; D000072417:Protein Domains; D053245:RANK Ligand / Q000494:pharmacology; D053246:Receptor Activator of Nuclear Factor-kappa B / Q000737:chemistry* / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects; D057127:Single-Chain Antibodies / Q000737:chemistry / Q000276:immunology / Q000494:pharmacology*; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects
chemical_list: D000941:Antigens; D016328:NF-kappa B; D053245:RANK Ligand; D053246:Receptor Activator of Nuclear Factor-kappa B; D057127:Single-Chain Antibodies; D020928:Mitogen-Activated Protein Kinases
keywords: Cell differentiation; Chimeric protein; Osteoclast; Signal transduction; Signalobody