Loss of protein kinase C-δ protects against LPS-induced osteolysis owing to an intrinsic defect in osteoclastic bone resorption

PLoS One. 2013 Aug 8;8(8):e70815. doi: 10.1371/journal.pone.0070815. eCollection 2013.

Abstract

Bone remodeling is intrinsically regulated by cell signaling molecules. The Protein Kinase C (PKC) family of serine/threonine kinases is involved in multiple signaling pathways including cell proliferation, differentiation, apoptosis and osteoclast biology. However, the precise involvement of individual PKC isoforms in the regulation of osteoclast formation and bone homeostasis remains unclear. Here, we identify PKC-δ as the major PKC isoform expressed among all PKCs in osteoclasts; including classical PKCs (-α, -β and -γ), novel PKCs (-δ, -ε, -η and -θ) and atypical PKCs (-ι/λ and -ζ). Interestingly, pharmacological inhibition and genetic ablation of PKC-δ impairs osteoclastic bone resorption in vitro. Moreover, disruption of PKC-δ activity protects against LPS-induced osteolysis in mice, with osteoclasts accumulating on the bone surface failing to resorb bone. Treatment with the PKC-δ inhibitor Rottlerin, blocks LPS-induced bone resorption in mice. Consistently, PKC-δ deficient mice exhibit increased trabeculae bone containing residual cartilage matrix, indicative of an osteoclast-rich osteopetrosis phenotype. Cultured ex vivo osteoclasts derived from PKC-δ null mice exhibit decreased CTX-1 levels and MARKS phosphorylation, with enhanced formation rates. This is accompanied by elevated gene expression levels of cathepsin K and PKC -α, -γ and -ε, as well as altered signaling of pERK and pcSrc416/527 upon RANKL-induction, possibly to compensate for the defects in bone resorption. Collectively, our data indicate that PKC-δ is an intrinsic regulator of osteoclast formation and bone resorption and thus is a potential therapeutic target for pathological osteolysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetophenones / therapeutic use
  • Animals
  • Benzopyrans / therapeutic use
  • Bone Resorption / drug therapy
  • Bone Resorption / genetics
  • Bone Resorption / immunology
  • Bone Resorption / pathology
  • Cathepsin K / genetics
  • Cells, Cultured
  • Enzyme Inhibitors / therapeutic use
  • Female
  • Gene Deletion
  • Gene Expression Regulation
  • Lipopolysaccharides / immunology*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Osteoclasts / drug effects
  • Osteoclasts / immunology
  • Osteoclasts / metabolism
  • Osteoclasts / pathology*
  • Osteolysis / drug therapy
  • Osteolysis / genetics*
  • Osteolysis / immunology*
  • Osteolysis / pathology
  • Phosphorylation
  • Protein Kinase C-delta / antagonists & inhibitors*
  • Protein Kinase C-delta / genetics*
  • Protein Kinase C-delta / immunology
  • Signal Transduction

Substances

  • Acetophenones
  • Benzopyrans
  • Enzyme Inhibitors
  • Lipopolysaccharides
  • rottlerin
  • Prkcd protein, mouse
  • Protein Kinase C-delta
  • Cathepsin K

Grants and funding

This work was funded in part by grants from the National Health and Medical Research Council (number 5138323). This study was also supported by a grant from the National Natural Science Foundation of China (NSFC) (number 81228013). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.