Identification of glycolytic pathway as RUNX1/ETO-dependent for propagation and survival

Introduction:

RUNX1/ETO is an oncogenic fusion protein, results from t(8;21) and accounts for 10 – 12% AML cases. The fusion protein is required for maintaining the leukaemic phenotype including both leukaemic clonogenicity and growth.

Method:

To further characterise the role of those RUNX1/ETO maintained genes in leukaemia progression we performed both an in vitro and an in vivo shRNA screen targeting those genes.

Result:

Functional characterisation of RUNX1/ETO (RE) direct targets and RNAi screen have identified genes related to glycolysis as potential required factors for leukaemic cells propagation. Notably, SLC2A3 only scored in vivo but not in vitro under standard tissue culture conditions, while PFKP scored in both settings. However, downregulation of SLC2A3 leads to a significantly decreased proliferation in limited glucose and oxygen content. PFKP knockdown results in loss of populations in all conditions. Furthermore, double knockdown of SLC2A3 and PFKP causes G1 cell cycle arrest and has a greater effect in reducing RE-leukaemia cells compared to single knockdown. Inhibition of glycolysis by 2-deoxyglucose also shows RUNX1/ETO-expressing cells are more susceptible to this inhibitor and induce apoptosis associated with overall decrease in cell number.

Conclusion:

Collectively, we have demonstrated that t(8;21) positive cells are dependent on glycolysis, which may instruct new treatment strategies with reduced toxicity.