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. 2014 Dec;28(12):2403-6.
doi: 10.1038/leu.2014.235. Epub 2014 Jul 31.

Primary acute myeloid leukemia cells with IDH1 or IDH2 mutations respond to a DOT1L inhibitor in vitro

S M Sarkaria  1 , M J Christopher  1 , J M Klco  2 , T J Ley  1
Affiliations

Primary acute myeloid leukemia cells with IDH1 or IDH2 mutations respond to a DOT1L inhibitor in vitro

S M Sarkaria et al. Leukemia. 2014 Dec.
No abstract available

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Conflict of interest statement

CONFLICT OF INTEREST

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1. EPZ004777 Alters Growth and Differentiation of Primary AML cells with MLL rearrangements
(A) Impact of EPZ004777 treatment on the growth of primary AML cells with (top panels) and without MLL rearrangements (bottom panels). Absolute cell numbers on y-axis; note different scales due to different rates of growth. Cells were expanded using a previously described stromal co-culture technique (also see methods in supplemental material) in the presence of DMSO or increasing concentrations of EPZ004777 (0.1 µM, 1 µM, 10 µM) over a 10-day period. Data represent mean values from two experiments assayed in duplicate ± SD (error bars). (B) Representative growth curve of MLL-ELL primary cells treated with increasing concentrations of EPZ004777 (left panel). Summary graph of cell growth results from two MLL-ELL samples treated with different doses of EPZ004777 (right panel); results are plotted as percent relative to DMSO control. Data represent mean values from two experiments assayed in duplicate ± SD (left) or SEM (right). (C) Histograms depicting the cell surface expression of myeloid markers CD11b (left panels) and CD14 (right panels) in two representative patient samples (one with a MLL translocation and one without). Cells were incubated in the presence of 10 µM EPZ004777 for 10 days and analyzed by flow cytometry. (D) Wright-Giemsa-stained cytospins of representative MLL and non-MLL rearranged patient samples treated for 10 days with DMSO or 10 µM EPZ004777. Scale bars represent 20 µm.
Figure 2
Figure 2. EPZ004777 Impairs the Growth and Differentiation of Genetically Defined, Non-MLL Rearranged Primary AML Cells
(A) Growth curves of primary cells with an IDH1 mutation (UPN 807970), a MLL-PTD mutation (UPN 817156), both mutations (UPN 914247), or neither mutation (UPN 868442) incubated in the presence of DMSO or increasing concentrations of EPZ004777 (0.1 µM, 1 µM, 10 µM) over a 10-day period. Refer to Supplementary Table S1 for more detailed information on individual primary samples. Absolute cell numbers on y-axis; note different scales due to different rates of growth. Cells were expanded using the stromal co-culture technique (see methods). Data represent mean values from two experiments assayed in duplicate ± SD (error bars). (B) Impact of EPZ004777 on the proliferation of IDH wildtype (WT; UPN 868442 & 721214) and IDH mutant (all others) primary cells; refer to Supplementary Table S1 for more details on individual samples. Results represent number of viable cells counted after 10 days, and are expressed as a percentage relative to DMSO controls, to adjust for differences in growth rates between different primary samples. Data represent mean values from two experiments assayed in duplicate ± SEM (error bars). (C) Cell surface expression of CD11b (left panels) and CD14 (right panels) in two representative IDH mutant patient samples. Cells were incubated in the presence of 10 µM EPZ004777 for 10 days and analyzed by flow cytometry. (D) Wright-Giemsa-stained cytospins of representative IDH wildtype and mutant patient samples treated for 10 days with DMSO or 10 µM EPZ004777. Scale bars represent 20 µm.
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