sirolimus and geranylgeraniol

Multinucleated bone-resorbing osteoclasts (Ocl) are cells of hematopoietic origin that play a major role in osteoporosis pathophysiology. Ocl survival and activity require M-CSF and RANK ligand (RANKL). M-CSF signals to Akt, while RANKL, like TNFalpha, activates NF-kappaB. We show here that although these are separate pathways in the Ocl, signaling of all three cytokines converges on mammalian target of rapamycin (mTOR) as part of their antiapoptotic action. Accordingly, rapamycin blocks M-CSF- and RANKL-dependent Ocl survival inducing apoptosis, and suppresses in vitro bone resorption proportional to the reduction in Ocl number. The cytokine signaling intermediates for mTOR/ribosomal protein S6 kinase (S6K) activation include phosphatidylinositol-3 kinase, Akt, Erks and geranylgeranylated proteins. Inhibitors of these intermediates suppress cytokine activation of S6K and induce Ocl apoptosis. mTOR regulates protein translation acting via S6K, 4E-BP1 and S6. We find that inhibition of translation by other mechanisms also induces Ocl apoptosis, demonstrating that Ocl survival is highly sensitive to continuous de novo protein synthesis. This study thus identifies mTOR/S6K as an essential signaling pathway engaged in the stimulation of cell survival in osteoclasts.

Topics: Acid Phosphatase; Actins; Adaptor Proteins, Signal Transducing; Alendronate; Alkyl and Aryl Transferases; Animals; Apoptosis; Blotting, Western; Bone Marrow Cells; Bone Resorption; Carrier Proteins; Caspase 3; Caspase 9; Caspases; Cell Cycle Proteins; Cell Survival; Collagen; Collagen Type I; Cytokines; Diterpenes; Enzyme Inhibitors; Eukaryotic Initiation Factors; Immunohistochemistry; Interleukin-1; Isoenzymes; Macrophage Colony-Stimulating Factor; Membrane Glycoproteins; Mice; Models, Biological; NF-kappa B; Osteoclasts; Peptides; Phosphoproteins; Protein Kinases; Protein Serine-Threonine Kinases; Protein Synthesis Inhibitors; Proto-Oncogene Proteins; Proto-Oncogene Proteins c-akt; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Ribosomal Protein S6; Ribosomal Protein S6 Kinases, 70-kDa; Serine-Threonine Kinase 3; Signal Transduction; Sirolimus; Tartrate-Resistant Acid Phosphatase; TOR Serine-Threonine Kinases; Tumor Necrosis Factor-alpha