Fig. 1: Treatment with Menin inhibitor ziftomenib depletes Menin expression, alters MLL1 target gene expressions in patient-derived AML stem cells defined by high expression of CLEC12A, CD123, CD99, and CD33 and induces differentiation and cell death of AML cells with MLL1 rearrangement or mutant NPM1. | Leukemia

Fig. 1: Treatment with Menin inhibitor ziftomenib depletes Menin expression, alters MLL1 target gene expressions in patient-derived AML stem cells defined by high expression of CLEC12A, CD123, CD99, and CD33 and induces differentiation and cell death of AML cells with MLL1 rearrangement or mutant NPM1.

From: Activity of menin inhibitor ziftomenib (KO-539) as monotherapy or in combinations against AML cells with MLL1 rearrangement or mutant NPM1

Fig. 1

A MOLM13 cells were treated with the indicated concentrations of ziftomenib for 7 days. Following this, the % of CD11b-positive cells was determined by flow cytometry. Features of morphologic differentiation were assessed in cells cytospun onto glass slides and stained with hematoxylin and eosin. Columns; mean of three experiments ± S.E.M. B OCI-AML3, MV4-11, MOLM13, THP1 and NOMO-1 cells were treated with the indicated concentrations of ziftomenib for 96 h. At the end of treatment, cells were stained with To-Pro-3 iodide and the percentage of non-viable cells was determined by flow cytometry. Columns; mean of three experiments ± S.E.M. C Representative immunoblot analyses of Menin and MLL1 fusion target genes in MOLM13 and OCI-AML3 cells treated with the indicated concentrations of ziftomenib for 48 h. The expression of GAPDH in the lysates served as the loading control. D Isogenic MOLM13 cells with TP53 mutation (R175H or R248Q) or bi-allelic TP53 knockout (KO) were treated with the indicated concentrations of ziftomenib for 96 h. At the end of treatment, cells were stained with To-Pro-3 iodide and the percentage of non-viable cells was determined by flow cytometry. Columns; mean of three experiments ± S.E.M. *Cell death values significantly greater (p < 0.05) in MOLM13 TP53-R175H compared to MOLM13 (TP53 wt). +Cell death values significantly greater (p < 0.05) in MOLM13 TP53-KO compared to MOLM13 (TP53 wt). E Patient-derived (PD) AML cells with MLL1 rearrangement or with mtNPM1 expression were treated with the indicated concentrations of ziftomenib for 72 h. Following this, the cells were stained with To-Pro-3 iodide and the percentage of non-viable cells was determined by flow cytometry. Columns; mean ± S.E.M for each type of AML. *p < 0.05, ***p < 0.005, relative to untreated cells, as determined by a two-tailed, unpaired t-test. F Patient-derived MLL1-rearranged AML cells were treated with the indicated concentrations of ziftomenib for 48 h. Total cell lysates were prepared and immunoblot analyses were conducted. The expression levels of GAPDH in the cell lysates served as the loading control. G, H Patient-derived MLL1-rearranged and mtNPM1-expressing AML cells were treated with 1 µM of ziftomenib for 16 h. Then, cells were incubated with cocktails of rare, heavy metal ion-tagged antibodies against extracellular and intracellular proteins. Mass cytometry (CyTOF) analysis was performed on the untreated and treated cells and the data were analyzed by Astrolabe. Protein expression alterations in ziftomenib-treated cells are shown as a fold-change, relative to the control cells in phenotypically defined AML stem cells.

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