title: Overexpression of the granulocyte colony-stimulating factor gene impairs bone morphogenetic protein responsiveness in mice.
abstract: Granulocyte colony-stimulating factor (G-CSF) is the major hematopoietic growth factor regulating the production and differentiation of neutrophils. We previously demonstrated that permanent overexpression of G-CSF in transgenic mice produces a dramatic enlargement of the bone cavity and reduction of bone mass. This phenotype was shown to be associated with an increase of osteoclast-mediated bone resorption. As a way of determining the role of G-CSF in bone formation in vivo, an ectopic bone was induced subcutaneously into G-CSF transgenic mice by bone morphogenetic protein (BMP)-2, a potent initiator of bone and cartilage from undifferentiated mesenchymal cells. A BMP-2/atelocollagen pellet containing recombinant human BMP-2 was implanted into a dorsal subfascial pocket. At one week after implantation, proliferation of mesenchymal cells around the implant was significantly decreased in transgenic mice compared with control mice. At three weeks, an ectopic bone containing bone marrow was formed both in transgenic and control mice. However, the ectopic bones of the transgenic mice were smaller and less consistent than those of control mice, and the calcium contents were reduced to 56.2% of those of controls. The ectopic bone in the G-CSF mice showed poor development of both lamellar and trabecular bone. Semiquantitative reverse transcription-polymerase chain reaction analysis of the ectopic bone at 3 weeks disclosed no significant differences in the mRNA levels of type I collagen, osteopontin, and osteocalcin between G-CSF mice and control mice. Immunohistochemical study in G-CSF mice showed reduced staining of osteocalcin in the bone matrix surrounding the reduced number of osteoblasts. The half-life of BMP in the implants was prolonged to 7 to 9 days in the G-CSF mice, whereas it was 5 days in the control mice. Collectively, the permanent expression of G-CSF may retard the differentiation process of osteoblasts by impairing the initial induction of mesenchymal cells, resulting in reduction of bone mass, suggesting that G-CSF regulates the bone metabolism by modulating both osteoclast and osteoblast function. Furthermore, it is suggested that G-CSF is a potent modulator of the BMP-2 signal pathway in vivo.
authors.affiliation: Department of Orthopedic Surgery, Sapporo Medical University, Sapporo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kuwabara
authors.forename: T
authors.initials: T
authors.lastname: Wada
authors.forename: T
authors.initials: T
authors.lastname: Oda
authors.forename: H
authors.initials: H
authors.lastname: Yoshikawa
authors.forename: N
authors.initials: N
authors.lastname: Sawada
authors.forename: Y
authors.initials: Y
authors.lastname: Kokai
authors.forename: S
authors.initials: S
authors.lastname: Ishii
mesh_terms: D000818:Animals; D001846:Bone Development; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D001842:Bone and Bones / Q000033:anatomy & histology / Q000000981:diagnostic imaging / Q000378:metabolism; D002118:Calcium / Q000032:analysis; D020219:Chondrogenesis; D003094:Collagen / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism; D004343:Drug Implants; D005347:Fibroblasts / Q000502:physiology; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics* / Q000502:physiology*; D051379:Mice; D008822:Mice, Transgenic; D020556:Neutrophil Infiltration; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D053495:Osteopontin; D012333:RNA, Messenger / Q000096:biosynthesis; D011859:Radiography; D012795:Sialoglycoproteins / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta; D019076:Transgenes
chemical_list: C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D004343:Drug Implants; D012333:RNA, Messenger; C506895:SPP1 protein, human; D012795:Sialoglycoproteins; C506896:Spp1 protein, mouse; D016212:Transforming Growth Factor beta; C059821:atelocollagen; D015675:Osteocalcin; D053495:Osteopontin; D016179:Granulocyte Colony-Stimulating Factor; D003094:Collagen; D002118:Calcium