Effect of sgRNA-mediated disruption of GLUT1 on gene expression on c-Kit+ MLL::AF9 leukemia cells

Acute myeloid leukemia (AML) is initiated and propagated by leukemia stem cells (LSCs), a self-renewing population of leukemia cells. Here we performed an in vivo CRISPR screen and identified the facilitated glucose transporter type 1 (GLUT1) as a critical metabolic dependency for murine MLL::AF9 LSCs. GLUT1 disruption by genetic ablation or by pharmacological inhibition with BAY-876 led to suppression of leukemia progression and improved survival in an MLL::AF9 mouse model of AML. Glut1 inhibition further resulted in glycolytic suppression, decreased levels of tricarboxylic cycle intermediates, and elevated levels of amino acids. These changes in metabolic profile coincided with increased autophagic activity and differentiation of AML cells. Notably, dual inhibition of GLUT1 and oxidative phosphorylation exhibited synergistic anti-leukemic effects in human AML cells. Collectively, these findings increase our understanding of how murine LSCs are metabolically regulated and highlight GLUT1 inhibition as a promising therapeutic adjuvant approach in AML. Overall design: Mouse MLL-AF9 leukemia cells were transduced with control or Glut1 sgRNAs. Cells were collected 3 days post transduction and RNA sequencing was performed on FACS sorted GFP+ cells. Total number of samples - 12. Replicates - Control_sgRNA - 4, Glut1_sgRNA_1 - 4, Glut1_sgRNA_2 - 4.