Serine biosynthesis is a metabolic vulnerability in FLT3-ITD-driven acute myeloid leukaemia [iFLT3-ITD-Depletion_In VIVO]

We performed 3'-RNA-Sequencing on RNA isolated from murine MLL-AF9/iFLT3-ITD cells where the inducible iFLT3-ITD transgene had been depleted for 48 hours compared to non-depleted conditions in vivo to perform differential gene expression analysis. Overall design: 1 million MLL-AF9/iFLT3-ITD cells were transplanted into NSG mice via tail vein IV injection. Mice were fed doxycycline-containing chow and water to maintain inducible iFLT3-ITD transgene expression, and leukaemias were allowed to engraft for 96 hours. Half of the cohort of mice (n=3) was then placed on regular chow and water to deplete inducible iFLT3-ITD transgene expression for 48 hours, while the other half of the cohort (n=3) remained on doxycycline-containing chow and water to maintain inducible iFLT3-ITD transgene expression as a control. Mice were all culled after 48 hours and leukaemic cells sorted from the bone marrow. Total RNA was isolated from cells and 3'-RNA libraries prepared for next-generation sequencing.