Normal bone growth and remodeling are, in part, dependent upon factors that influence the number and activity of osteoclasts. Skeletal pathologies may result from alterations in the osteoclast plasma membrane or in signals that govern osteoclast cytodifferentiation and resorptive activity. Despite substantial recent insights into understanding the hematopoietic lineage and the interactions of local and systemic modulators of osteoclast development and function, many questions remain. In addition, the biochemical and molecular profile of the mature osteoclast is incomplete. The hypothesis upon which this application is based proposes that osteoclast development entails specific cell surface alterations and that discrete membrane-associated molecules are involved in osteoclast regulation and function.
The specific aims of this proposal are to: 1) further characterize the antigen reactive with the 121F monoclonal antibody (MAb) and to clarify its relationship with superoxide dismutase, 2) continue to identify and biochemically describe distinct osteoclast plasma membrane molecules recognized by a panel of osteoclast-specific MAbs, 3) investigate cell surface changes associated with osteoclast development and functional state, relative to the effects of known modulators of osteoclast development and activity (such as estradiol, calcitonin, interleukins 1 and 6, and TNF) and to the induced expression of protooncogenes (c-fos, c-jun, and c-src), and 4) identify osteoclast precursors and define stages of osteoclast cytodifferentiation using-these cell surface markers and molecular probes. Highly purified chick osteoclasts and a human leukemic cell line (FLG 29.1) will be used as sources for chicken and human antigens. In other experiments, marrow mononuclear cells which fuse to form multinucleated osteoclast-like giant cells in culture will serve as a developmental system in which to investigate the influence of known bone modulators on antigen expression and resorptive capability, and to define osteoclast precursors. Biochemical,physiological, immunohistochemical, and molecular cloning techniques will be employed to ascertain the nature and functional role of these antigens. Western and northern blot analyses will be used to examine their relationship to protooncogene expression. These results should not only expand our basic understanding of the cellular and molecular biology of the osteoclast, but also potentially aid in devising appropriate preventative and therapeutic strategies for the skeletal osteopenia of arthritis, osteoporosis, periodontal disease, and other local or metabolic disorders of bone.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
2R01AR032927-09
Application #
3156439
Study Section
Orthopedics and Musculoskeletal Study Section (ORTH)
Project Start
1984-07-01
Project End
1996-06-30
Budget Start
1992-07-01
Budget End
1993-06-30
Support Year
9
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Collin-Osdoby, Patricia; Osdoby, Philip (2012) Isolation and culture of primary chicken osteoclasts. Methods Mol Biol 816:119-43
Collin-Osdoby, Patricia; Osdoby, Philip (2012) RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Biol 816:187-202
Collin-Osdoby, Patricia; Yu, Xuefeng; Zheng, Hong et al. (2003) RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Med 80:153-66
Collin-Osdoby, Patricia; Anderson, Fred; Osdoby, Philip (2003) Primary isolation and culture of chicken osteoclasts. Methods Mol Med 80:65-88
Yu, Xuefeng; Huang, Yuefang; Collin-Osdoby, Patricia et al. (2003) Stromal cell-derived factor-1 (SDF-1) recruits osteoclast precursors by inducing chemotaxis, matrix metalloproteinase-9 (MMP-9) activity, and collagen transmigration. J Bone Miner Res 18:1404-18
Collin-Osdoby, Patricia; Rothe, Linda; Bekker, Simon et al. (2002) Basic fibroblast growth factor stimulates osteoclast recruitment, development, and bone pit resorption in association with angiogenesis in vivo on the chick chorioallantoic membrane and activates isolated avian osteoclast resorption in vitro. J Bone Miner Res 17:1859-71
Collin-Osdoby, P; Rothe, L; Anderson, F et al. (2001) Receptor activator of NF-kappa B and osteoprotegerin expression by human microvascular endothelial cells, regulation by inflammatory cytokines, and role in human osteoclastogenesis. J Biol Chem 276:20659-72
Collin-Osdoby, P; Rothe, L; Bekker, S et al. (2000) Decreased nitric oxide levels stimulate osteoclastogenesis and bone resorption both in vitro and in vivo on the chick chorioallantoic membrane in association with neoangiogenesis. J Bone Miner Res 15:474-88
Collin-Osdoby, P; Li, L; Rothe, L et al. (1998) Inhibition of avian osteoclast bone resorption by monoclonal antibody 121F: a mechanism involving the osteoclast free radical system. J Bone Miner Res 13:67-78
Khalkhali-Ellis, Z; Collin-Osdoby, P; Li, L et al. (1997) A human homolog of the 150 kD avian osteoclast membrane antigen related to superoxide dismutase and essential for bone resorption is induced by developmental agents and opposed by estrogen in FLG 29.1 cells. Calcif Tissue Int 60:187-93

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