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We also used the FMH9 line, derived from primary HoxA9/Meis1-transformed bone marrow progenitors, as an AML model.

In addition to demonstrating the functional relevance for FLT3-ITD-induced AML, this AML model has a number of important features.

MV4-11 cells were used for more in-depth analysis of the combination, as this well-established AML model can be used both in vitro and in vivo.

These results demonstrate that Bmi-1 is also required for self renewal of leukemic stem cells in the murine AML model.

The establishment of this preclinical AML model is of special relevance and significance to drug-sensitivity studies as in vitro cell culture-based screens do not accurately reflect in vivo effects and responses.

We screened genome-wide for aberrant CpG island methylation in three disease stages of a murine AML model that is driven by hypomorphic expression of the hematopoietic transcription factor PU.1.

The synergy translated in vivo in two different AML models, the SET-2 megakaryoblastic AML mouse model carrying a JAK2V617F mutation, and the MOLM-13 model of FLT3-ITD-driven AML.

Furthermore, many AML models are based on reciprocal chromosomal translocations that only reflect the minority of AML patients, whereas more than 50% of patients have a normal karyotype.

In accordance with its role in mouse AML models, the lentiviral vector-delivered JMJD3 expression-cassette resulted in JMJD3 overexpression and decreased the colony-forming potential of the primary leukemic blasts in six out of seven cases (in one M2b, one M3, two M4, and one unclassified AML case but only in one out of two M5 cases) (Fig. 3a and Supplementary Fig. 2a).

Activation of the abdominal HoxA-cluster genes is known to contribute to the leukaemogenic phenotype in our AML models.

Taken together, we hypothesize that leukaemogenic progression in MLL-ENL and MOZ-TIF2 AML models is associated with altered expression of HSPC-TFs which, in turn, is accompanied by corresponding changes in H3K9ac profiles at their candidate regulatory regions.