Hypermethylation of CXCR4 promoter in CD34+ cells from patients with primary myelofibrosis

Costanza Bogani, Vanessa Ponziani, Paola Guglielmelli, Cristophe Desterke, Vittorio Rosti, Alberto Bosi, Marie Caroline Le Bousse-Kerdilès, Giovanni Barosi, Alessandro M. Vannucchi

Research output: Contribution to journalArticlepeer-review

Abstract

Constitutive mobilization of CD34+ cells in patients with primary myelofibrosis (PMF) has been attributed to proteolytic disruption of the CXCR4/SDF-1 axis and reduced CXCR4 expression. We document here that the number of circulating CD34+/CXCR4+ cells in PMF patients, as well as the cellular CXCR4 expression, was directly related to CXCR4 mRNA level and that reduced CXCR4 mRNA level was not due to SDF-1-induced downregulation. To address whether epigenetic regulation contributes to defective CXCR4 expression, we studied the methylation status of the CXCR4 promoter using methylation-specific polymerase chain reaction and methylation-specific sequencing in the JAK2V617F-positive HEL cell line and in CD34+ cells. We found that CD34+ cells from PMF patients, unlike those from normal subjects, presented hypermethylation of CXCR4 promoter CpG island 1. Following incubation with the demethylating agent 5-Aza-2′-deoxycytidine (5-AzaD), the percentage of PMF CD34+ cells expressing CXCR4 increased 3-10 times, whereas CXCR4 mRNA level increased approximately 4 times. 5-AzaD-treated PMF CD34+ cells displayed almost complete reversal of CpG1 island 1 hypermethylation and showed enhanced migration in vitro in response to SDF-1. These data point to abnormal methylation of the CXCR4 promoter as a mechanism contributing to constitutive migration of CD34 + cells in PMF.

Original languageEnglish
Pages (from-to)1920-1930
Number of pages11
JournalStem Cells
Volume26
Issue number8
DOIs
Publication statusPublished - Aug 2008

Keywords

  • CD34+ cell
  • CXCR4
  • Epigenetic
  • Methylation
  • Myelofibrosis

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Molecular Medicine

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