The Retinal Determination Gene Network can confer chemoresistant properties to AML

HoxA9, Six1 and Eya1 are targets of MOZ-TIF2 and MLL-AF9 fusion proteins in murine models. Hoxa9 overexpression can induce AML in murine HSPCs with a latency of ~6-9 months. Co-overexpression of RDGN members Six1 or Eya1 with Hoxa9 can enhance the differentiation block and colony forming potential of HoxA9 alone in vitro. RNA-seq was carried out on these lines to establish genetic pathways responsible for these chages in leukemogenicity. Benzarone is a potent EYA1 inhibitor. We wanted to assess if this drug was able to reverse the leukemogenic properties that EYA1 adds to HOXA9. Overall design: Murine c-Kit+ cells were isolated from the bone marrow of female C57BL/6 mice. Cells were maintained in MT2 media for 1 day before being retrovirally transduced with flourescent reporter plasmids encoding HoxA9 alone, or together with Six1/Eya1. Lines were established in methylcellulose culture for 3 weeks before being cell sorted for their reporter construct and squenced. HOXA9_EYA1 lines were treated with either 5ug Benzarone or a DMSO (vehicle) control for 72hrs before RNA was extracted and samples sent for sequencing. HOXA9 alone (4 samples), HOXA9+SIX1 (4 samples).