Cystathionine γ-lyase accelerates osteoclast differentiation: identification of a novel regulator of osteoclastogenesis by proteomic analysis

Arterioscler Thromb Vasc Biol. 2014 Mar;34(3):626-34. doi: 10.1161/ATVBAHA.113.302576. Epub 2013 Dec 19.


Objective: Clinical evidence has linked vascular calcification in advanced atherosclerotic plaques with overt cardiovascular disease and mortality. Bone resorbing monocyte-derived osteoclast-like cells are sparse in these plaques, indicating that their differentiation capability could be suppressed. Here, we seek to characterize the process of osteoclastogenesis by identifying novel regulators and pathways, with the aim of exploring possible strategies to reduce calcification.

Approach and results: We used a quantitative mass spectrometry strategy, tandem mass tagging, to quantify changes in the proteome of osteoclast-like cells differentiated from RAW264.7 cells in response to, receptor activator of nuclear factor κ-B ligand induction, a common in vitro model for osteogenesis. More than 4000 proteins were quantified, of which 138 were identified as novel osteoclast-related proteins. We selected 5 proteins for subsequent analysis (cystathionine γ-lyase [Cth/CSE], EGF-like repeat and discoidin I-like domain-containing protein 3, integrin α FG-GAP repeat containing 3, adseverin, and serpinb6b) and show that gene expression levels are also increased. Further analysis of the CSE transcript profile reveals an early onset of an mRNA increase. Silencing of CSE by siRNA and dl-propargylglycine, a CSE inhibitor, attenuated receptor activator of nuclear factor κ-B ligand-induced tartrate-resistant acid phosphatase type 5 activity and pit formation, suggesting that CSE is a potent inducer of calcium resorption. Moreover, knockdown of CSE suppressed expression of osteoclast differentiation markers.

Conclusions: Our large-scale proteomics study identified novel candidate regulators or markers for osteoclastogenesis and demonstrated that CSE may act in early stages of osteoclastogenesis.

Keywords: RANKL protein; macrophages; osteoclast; proteomics; vascular calcification.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkynes / pharmacology
  • Animals
  • Aorta, Thoracic / metabolism
  • Apolipoproteins E / deficiency
  • Biomarkers
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / drug effects
  • Cell Differentiation
  • Cell Line, Tumor
  • Cystathionine gamma-Lyase / antagonists & inhibitors
  • Cystathionine gamma-Lyase / genetics
  • Cystathionine gamma-Lyase / physiology*
  • Dietary Fats / toxicity
  • Gene Expression Profiling
  • Glycine / analogs & derivatives
  • Glycine / pharmacology
  • Macrophage Colony-Stimulating Factor / pharmacology
  • Macrophages / cytology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • NF-kappa B / metabolism
  • Osteoclasts / cytology
  • Osteoclasts / enzymology*
  • Osteoclasts / metabolism
  • Proteomics
  • RANK Ligand / pharmacology
  • RNA Interference
  • RNA, Messenger / biosynthesis
  • RNA, Small Interfering / pharmacology
  • Spectrometry, Mass, Electrospray Ionization
  • Transcription, Genetic


  • Alkynes
  • Apolipoproteins E
  • Biomarkers
  • Dietary Fats
  • NF-kappa B
  • RANK Ligand
  • RNA, Messenger
  • RNA, Small Interfering
  • Tnfsf11 protein, mouse
  • propargylglycine
  • Macrophage Colony-Stimulating Factor
  • Cystathionine gamma-Lyase
  • Glycine