Functional characterisation of SLC2A3 and PFKP in t(8;21)-positive acute myeloid leukemia

Abstract

The translocation t(8;21) is the most prevalent chromosomal rearrangement in acute myeloid leukaemia (AML) and results in the expression of the oncogenic fusion protein RUNX1/ETO. The fusion protein is required for maintaining the leukaemic phenotype. Previous in vivo work identified genes related to glycolysis, namely SLC2A3 and PFKP as potential transcriptional targets for RUNX1/ETO. This project aimed to functionally investigate the role of SLC2A3 and PFKP in RUNX1/ETO-driven leukaemia both in vivo and in vitro. SLC2A3 significantly maintained the fitness of leukaemic cells under limited glucose and hypoxic conditions in the RUNX1/ETO-expressing Kasumi-1 cell line. Notably, SLC2A3 knockdown impaired the competitive fitness of patient-derived xenograft (PDX) cells in high glucose and oxygen condition. Furthermore, PFKP knockdown results in loss of populations both in vitro and in vivo. Besides, double knockdown of SLC2A3 and PFKP caused G1 cell cycle arrest and had a greater effect in impairing leukaemic cell propagation and fitness when compared to the single knockdowns. Moreover, RUNX1/ETO-expressing cells showed to have a high sensitivity towards 2- deoxyglucose (2DG). 2DG is an analogue of glucose molecule that blocks glycolysis upstream of PFKP. RUNX1/ETO cells treated with this inhibitor undergo apoptosis associated with the overall decrease in cell number. The combination of 2DG and cell cycle inhibitor, palbociclib led to an additive effect and severely impaired expressions of genes in the glycolysis pathway. Collectively, the oncogenic fusion protein, RUNX1/ETO shows dependency on glycolysis through SLC2A3 and PFKP. Targeting these two genes may instruct new treatment strategies with reduced toxicity in t(8;21) leukaemia.