TY - JOUR
T1 - Hypermethylation of CXCR4 promoter in CD34+ cells from patients with primary myelofibrosis
AU - Bogani, Costanza
AU - Ponziani, Vanessa
AU - Guglielmelli, Paola
AU - Desterke, Cristophe
AU - Rosti, Vittorio
AU - Bosi, Alberto
AU - Le Bousse-Kerdilès, Marie Caroline
AU - Barosi, Giovanni
AU - Vannucchi, Alessandro M.
PY - 2008/8
Y1 - 2008/8
N2 - 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.
AB - 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.
KW - CD34+ cell
KW - CXCR4
KW - Epigenetic
KW - Methylation
KW - Myelofibrosis
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U2 - 10.1634/stemcells.2008-0377
DO - 10.1634/stemcells.2008-0377
M3 - Article
C2 - 18511598
AN - SCOPUS:55049114480
VL - 26
SP - 1920
EP - 1930
JO - Stem Cells
JF - Stem Cells
SN - 1066-5099
IS - 8
ER -