BRD3/4 inhibition and FLT3-ligand deprivation target pathways that are essential for the survival of human MLL-AF9+ leukemic cells

PLoS One. 2017 Dec 14;12(12):e0189102. doi: 10.1371/journal.pone.0189102. eCollection 2017.

Abstract

In the present work we aimed to identify targetable signaling networks in human MLL-AF9 leukemias. We show that MLL-AF9 cells critically depend on FLT3-ligand induced pathways as well as on BRD3/4 for their survival. We evaluated the in vitro and in vivo efficacy of the BRD3/4 inhibitor I-BET151 in various human MLL-AF9 (primary) models and patient samples and analyzed the transcriptome changes following treatment. To further understand the mode of action of BRD3/4 inhibition, we performed ChIP-seq experiments on the MLL-AF9 complex in THP1 cells and compared it to RNA-seq data of I-BET151 treated cells. While we could confirm a consistent and specific downregulation of key-oncogenic drivers such as MYC and BCL2, we found that the majority of I-BET151-responsive genes were not direct MLL-AF9 targets. In fact, MLL-AF9 specific targets such as the HOXA cluster, MEIS1 and other cell cycle regulators such as CDK6 were not affected by I-BET151 treatment. Furthermore, we also highlight how MLL-AF9 transformed cells are dependent on the function of non-mutated hematopoietic transcription factors and tyrosine kinases such as the FLT3-TAK1/NF-kB pathway, again impacting on BCL2 but not on the HOXA cluster. We conclude that BRD3/4 and the FLT3-TAK1/NF-kB pathways collectively control a set of targets that are critically important for the survival of human MLL-AF9 cells.

MeSH terms

  • Cell Cycle Proteins
  • Cell Survival*
  • Cells, Cultured
  • Humans
  • Infant, Newborn
  • Leukemia / metabolism
  • Leukemia / pathology*
  • Nuclear Proteins / antagonists & inhibitors*
  • RNA-Binding Proteins / antagonists & inhibitors*
  • Transcription Factors / antagonists & inhibitors*
  • fms-Like Tyrosine Kinase 3 / metabolism*

Substances

  • BRD3 protein, human
  • BRD4 protein, human
  • Cell Cycle Proteins
  • Nuclear Proteins
  • RNA-Binding Proteins
  • Transcription Factors
  • FLT3 protein, human
  • fms-Like Tyrosine Kinase 3

Grants and funding

This work was supported by the EU (FP7-PEOPLE-2010-ITN-264361 EuroCancer StemCell Training network). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.