Targeting Integrin-β1 Impedes Cytokine-Induced Osteoclast Differentiation: A Potential Pharmacological Intervention in Pathological Osteolysis

Purpose: To examine whether integrin-β1 is essential for osteoclast differentiation and function and if it can be targeted for pharmacological intervention in pathological osteolysis.
Methods: Control and Integrin-β1 knockdown RAW 264.7 cells were treated with receptor activator of nuclear factor kappa-B (RANKL) or TNF-α and evaluated for osteoclast differentiation. Osteoclast differentiation and function were evaluated by marker protein analysis, tartrate-resistant acid phosphatase (TRAP) and resorption assays. Furthermore, downstream molecular signaling analysis was probed using small molecule inhibitors and blocking antibodies, and evaluated by immunoblotting.
Results: Integrin-β1 knockdown cells showed reduced osteoclast differentiation following TNF-α treatment while no change was seen after RANKL treatment (p < 0.05). Immunoblot-based molecular signaling analysis showed involvement of MAPK kinase signaling in mediating TNF-α/integrin-β1- induced osteoclastogenesis. Finally, when MAPK kinase inhibitor (2.5 and 5 μM; p < 0.05) and integrin- β1 blocking antibody (2.5 and 5 μg/mL; p < 0.05) was used to specifically attenuate TNF-α induced osteoclastogenesis, no change was observed in RANKL-induced osteoclast formation.
Conclusion: The data obtained highlight the role of integrin-β1 in TNF-α-induced osteoclastogenesis, but not in RANKL pathway. Given that, inflammatory cytokine secretions such as TNF-α are progressively implicated in pathological osteolysis, targeting this pathway may attenuate osteolysis in pathological bone tissues.

Keywords
Osteoclast differentiation; Integrin-β1; Receptor activator of nuclear factor kappa-B; TNF-alpha; Mitogen activated protein kinase; Cytokines; Skeletal disease