title: Pretreatment long interspersed element (LINE)-1 methylation levels, not early hypomethylation under treatment, predict hematological response to azacitidine in elderly patients with acute myeloid leukemia.
abstract: BACKGROUND
  Epigenetic modulations, including changes in DNA cytosine methylation, are implicated in the pathogenesis and progression of acute myeloid leukemia (AML). Azacitidine is a hypomethylating agent that is incorporated into RNA as well as DNA. Thus, there is a rationale to its use in patients with AML. We determined whether baseline and/or early changes in the methylation of long interspersed element (LINE)-1 or CDH13 correlate with bone marrow blast clearance, hematological response, or survival in patients with AML treated with azacitidine.
  
  
  METHODS
  An open label, phase I/II trial was performed in 40 AML patients (median bone marrow blast count was 42%) unfit for intensive chemotherapy treated with azacitidine 75 mg/m(2)/day subcutaneously for 5 days every 4 weeks. Bone marrow mononuclear cell samples were taken on day 0 (pretreatment) and day 15 during the first treatment cycle; LINE-1 and CDH13 methylation levels were quantified by methylation-specific, semiquantitative, real-time polymerase chain reaction.
  
  
  RESULTS
  Treatment with azacitidine significantly reduced LINE-1 but not CDH13 methylation levels over the first cycle (P < 0.0001). Absolute LINE-1 methylation levels tended to be lower on day 0 (P = 0.06) and day 15 of cycle 1 (P = 0.03) in patients who went on to achieve subsequent complete remission, partial remission or hematological improvement versus patients with stable disease. However, the decrease in LINE-1 methylation over the first treatment cycle did not correlate with subsequent response (P = 0.31). Baseline methylation levels of LINE-1 or CDH13 did not correlate with disease-related prognostic factors, including cytogenetic risk, relapsed/refractory AML, or presence of NPM1 or FLT3 mutations. No correlation was observed between LINE-1 or CDH13 methylation levels and overall survival.
  
  
  CONCLUSIONS
  Analysis of baseline LINE-1 methylation levels may help identify elderly AML patients who are most likely to respond to azacitidine therapy.
authors.affiliation: Division of Hematology and Oncology, University of Leipzig, Leipzig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Cross
authors.forename: Enrica
authors.initials: E
authors.lastname: Bach
authors.forename: Thao
authors.initials: T
authors.lastname: Tran
authors.forename: Rainer
authors.initials: R
authors.lastname: Krahl
authors.forename: Nadja
authors.initials: N
authors.lastname: Jaekel
authors.forename: Dietger
authors.initials: D
authors.lastname: Niederwieser
authors.forename: Christian
authors.initials: C
authors.lastname: Junghanss
authors.forename: Georg
authors.initials: G
authors.lastname: Maschmeyer
authors.forename: Haifa Kathrin
authors.initials: HK
authors.lastname: Al-Ali
keywords: CDH13; DNA methylation; LINE-1 methylation; acute myeloid leukemia; azacitidine; clinical response