Identification of DJ-1/PARK-7 as a determinant of stroma-dependent and TNF-alpha-induced apoptosis in MDS using mass spectrometry and phosphopeptide analysis

Blood. 2010 Mar 11;115(10):1993-2002. doi: 10.1182/blood-2009-08-236992. Epub 2010 Jan 8.

Abstract

In patients with myelodysplastic syndromes (MDS), apoptosis in hematopoietic cells is up-regulated in low-grade disease, whereas advanced disease is characterized by apoptosis resistance. We have shown that marrow stroma-derived signals convey sensitivity to tumor-necrosis-factor alpha (TNF-alpha)-mediated apoptosis in otherwise-resistant KG1a myeloid cells and CD34(+) cells from MDS marrow. Here, we used a PhosphoScan proteomic liquid chromatography-mass spectrometry method to identify signals relevant for this effect. The transcription factor DJ-1/PARK-7 (DJ-1) was highly phosphorylated in KG1a cells cultured without stroma but dephosphorylated after stroma coculture, whereas expression of p53 increased significantly, suggesting a stroma contact-dependent effect of DJ-1 on p53. In CD34(+) marrow cells from advanced MDS, expression of DJ-1 was up-regulated, whereas p53 levels were low, resulting in significantly greater DJ-1/p53 ratios than in patients with low-grade MDS (P = .01). DJ-1 levels were correlated with increasing International Prognostic Scoring System scores (P = .006). Increasing DJ-1/p53 ratios were associated with an increased risk of mortality, although the correlation did not reach statistical significance (P = .18). These data suggest that DJ-1/p53 interactions contribute to apoptosis resistance in clonal myeloid cells and may serve as a prognostic marker in patients with MDS.

Publication types

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

MeSH terms

  • Adult
  • Aged
  • Apoptosis / drug effects*
  • Apoptosis / physiology
  • Cells, Cultured
  • Female
  • Humans
  • Intracellular Signaling Peptides and Proteins / analysis
  • Intracellular Signaling Peptides and Proteins / chemistry
  • Intracellular Signaling Peptides and Proteins / metabolism
  • Intracellular Signaling Peptides and Proteins / physiology*
  • Male
  • Mass Spectrometry / methods
  • Middle Aged
  • Myelodysplastic Syndromes / genetics
  • Myelodysplastic Syndromes / metabolism
  • Myelodysplastic Syndromes / pathology*
  • Oncogene Proteins / analysis
  • Oncogene Proteins / chemistry
  • Oncogene Proteins / metabolism
  • Oncogene Proteins / physiology*
  • Phosphopeptides / analysis
  • Phosphopeptides / chemistry
  • Phosphorylation
  • Protein Deglycase DJ-1
  • Protein-Tyrosine Kinases / metabolism
  • Retrospective Studies
  • Stromal Cells / metabolism
  • Stromal Cells / physiology*
  • Tumor Necrosis Factor-alpha / pharmacology*
  • Tumor Suppressor Protein p53 / metabolism

Substances

  • Intracellular Signaling Peptides and Proteins
  • Oncogene Proteins
  • Phosphopeptides
  • Tumor Necrosis Factor-alpha
  • Tumor Suppressor Protein p53
  • Protein-Tyrosine Kinases
  • PARK7 protein, human
  • Protein Deglycase DJ-1