title: Epigenetic therapy of acute myeloid leukemia using 5-aza-2'-deoxycytidine (decitabine) in combination with inhibitors of histone methylation and deacetylation.
abstract: BACKGROUND
  The silencing of tumor suppressor genes (TSGs) by aberrant DNA methylation occurs frequently in acute myeloid leukemia (AML). This epigenetic alteration can be reversed by 5-aza-2'-deoxcytidine (decitabine, 5-AZA-CdR). Although 5-AZA-CdR can induce complete remissions in patients with AML, most patients relapse. The effectiveness of this therapy may be limited by the inability of 5-AZA-CdR to reactivate all TSGs due to their silencing by other epigenetic mechanisms such as histone methylation or chromatin compaction. EZH2, a subunit of the polycomb repressive complex 2, catalyzes the methylation of histone H3 lysine 27 (H3K27) to H3K27me3. 3-Deazaneplanocin-A (DZNep), an inhibitor of methionine metabolism, can reactivate genes silenced by H3K27me3 by its inhibition of EZH2. In a previous report, we observed that 5-AZA-CdR, in combination with DZNep, shows synergistic antineoplastic action against AML cells. Gene silencing due to chromatin compaction is attributable to the action of histone deacetylases (HDAC). This mechanism of epigenetic gene silencing can be reversed by HDAC inhibitors such as trichostatin-A (TSA). Silent TSGs that cannot be reactivated by 5-AZA-CdR or DZNep have the potential to be reactivated by TSA. This provides a rationale for the use of HDAC inhibitors in combination with 5-AZA-CdR and DZNep to treat AML.
  
  
  RESULTS
  The triple combination of 5-AZA-CdR, DZNep, and TSA induced a remarkable synergistic antineoplastic effect against human AML cells as demonstrated by an in vitro colony assay. This triple combination also showed a potent synergistic activation of several key TSGs as determined by real-time PCR. The triple combination was more effective than the combination of two agents or a single agent. Microarray analysis showed that the triple combination generated remarkable changes in global gene expression.
  
  
  CONCLUSIONS
  Our data suggest that it may be possible to design a very effective therapy for AML using agents that target the reversal of the following three epigenetic "lock" mechanisms that silence gene expression: DNA methylation, histone methylation, and histone deacetylation. This approach merits serious consideration for clinical investigation in patients with advanced AML.
authors.affiliation: Département de Pharmacologie, Université de Montréal, 2900 Édouard-Montpetit, Montréal, QC H3T 1J4, Canada ; Centre de recherche, Service d'hématologie/oncologie, CHU-Saint-Justine, Montréal, Québec H3T 1C5, Canada.
authors.forename: Richard L
authors.initials: RL
authors.lastname: Momparler
authors.affiliation: Centre de recherche, Service d'hématologie/oncologie, CHU-Saint-Justine, Montréal, Québec H3T 1C5, Canada.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Côté
authors.affiliation: Centre de recherche, Service d'hématologie/oncologie, CHU-Saint-Justine, Montréal, Québec H3T 1C5, Canada.
authors.forename: Louise F
authors.initials: LF
authors.lastname: Momparler
authors.affiliation: Department of Biology, New York University, Saadiyat Island, PO Box 129188, Abu Dhabi, United Arab Emirates.
authors.forename: Youssef
authors.initials: Y
authors.lastname: Idaghdour
keywords: 3-deazaneplanocin-A; 5-aza-2’-deoxcytidine; Decitabine; EZH2; Epigenetic therapy; Histone deacetylase inhibitors; Myeloid leukemia