Inflammatory osteolysis, as occurs in rheumatoid arthritis and orthopedic implant loosening, reflects accelerated osteoclast (OC) recruitment and activation. Hence, discovering the means by which local inflammation recruits and activates OCs is central to preventing this crippling complication. Our overriding goal has been to detail the mechanisms by which cytokines, such as TNF, RANKL and M-CSF mediate bone loss. We have achieved the aims of our previous application by characterizing the contributions of various TNF target cells to the osteoclastogenic process and detailing many of the intracellular events by which TNF promotes osteoclastogenesis. In the present proposal we turn to the cytokine essential for all pathological bone resorption, namely RANKL, whose crystal structure, in complex with its receptor, RANK, we resolved in the current funding period. While RANKL is essential for OC differentiation, it also enhances the bone resorptive activity of the mature OC, a process we find depends upon the cytokine's regulation of the cytoskeletal adaptor protein, paxillin and the small GTPase, Rac. Furthermore, our resolution of the RANKL/RANK crystal structure and that of RANKL with the cysteine-rich domains of OPG, the decoy receptor, positions us to develop structure-based RANKL antagonists to arrest RANK signaling in the OC. Given that inhibition of the RANK signaling pathway has proven therapeutic benefit, these novel RANKL inhibitors may impact the treatment of pathological osteolysis. Thus, we hypothesize that: 1) RANKL activation of the OC is mediated via paxillin 2) RANKL activation of the OC is mediated via Rac and 3) Structure-based RANKL antagonists will arrest OC development and function.
Our Specific Aims are therefore to 1) Determine the mechanisms by which paxillin mediates RANKL activation of the OC 2) Determine the mechanisms by which RANKL activates Rac in the OC and 3) Engineer variants of RANKL and OPG that disrupt RANK signaling in the OC.

Public Health Relevance

. Osteoclasts are the cells which destroy bone, and thus, their increased activity is responsible for most forms of pathological bone loss such as osteoporosis or that attending rheumatoid arthritis. The key molecule which activates ostoclasts is known as RANK ligand (RANKL). The purpose of this proposal is to gain insight into the mechanism by which RANKL activates osteoclasts and to design candidate drugs which will inhibit this event and consequently, prevent pathological bone loss.

National Institute of Health (NIH)
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Research Project (R01)
Project #
Application #
Study Section
Skeletal Biology Structure and Regeneration Study Section (SBSR)
Program Officer
Chen, Faye H
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Washington University
Schools of Medicine
Saint Louis
United States
Zip Code
Izawa, Takashi; Rohatgi, Nidhi; Fukunaga, Tomohiro et al. (2015) ASXL2 Regulates Glucose, Lipid, and Skeletal Homeostasis. Cell Rep 11:1625-37
Warren, Julia T; Zou, Wei; Decker, Corinne E et al. (2015) Correlating RANK ligand/RANK binding kinetics with osteoclast formation and function. J Cell Biochem 116:2476-83
Fukunaga, Tomohiro; Zou, Wei; Rohatgi, Nidhi et al. (2015) An insulin-sensitizing thiazolidinedione, which minimally activates PPAR?, does not cause bone loss. J Bone Miner Res 30:481-8
Zou, Wei; Teitelbaum, Steven L (2015) Absence of Dap12 and the ?v?3 integrin causes severe osteopetrosis. J Cell Biol 208:125-36
Warren, Julia T; Nelson, Christopher A; Decker, Corinne E et al. (2014) Manipulation of receptor oligomerization as a strategy to inhibit signaling by TNF superfamily members. Sci Signal 7:ra80
Fukunaga, Tomohiro; Zou, Wei; Warren, Julia T et al. (2014) Vinculin regulates osteoclast function. J Biol Chem 289:13554-64
Zhu, Tingting; Chappel, Jean C; Hsu, Fong-Fu et al. (2013) Type I phosphotidylinosotol 4-phosphate 5-kinase ? regulates osteoclasts in a bifunctional manner. J Biol Chem 288:5268-77
Zou, Wei; Croke, Monica; Fukunaga, Tomohiro et al. (2013) Zap70 inhibits Syk-mediated osteoclast function. J Cell Biochem 114:1871-8
Zou, Wei; Izawa, Takashi; Zhu, Tingting et al. (2013) Talin1 and Rap1 are critical for osteoclast function. Mol Cell Biol 33:830-44
Bush, Jason A; Kitaura, Hideki; Ma, Yuliang et al. (2012) Comparative proteomic analysis of a cytosolic fraction from ?3 integrin-deficient cells. Cancer Genomics Proteomics 9:1-13

Showing the most recent 10 out of 98 publications