Fig. 1: Identification and characterization of CM-444 and CM-1758, as a pan-HDACi, with high AML differentiation potencies at low non-cytotoxic doses.
From: Epigenetic-based differentiation therapy for Acute Myeloid Leukemia
A Epigenetic small-molecule screening was performed in HL-60 cell line treated daily with 25% GI50 of each compound for 48 h. CD11b and annexin-V were measured by flow cytometry. The data shown are the mean of three biologically independent experiments. B Percentage of inhibition of CM-444 and CM-1758 at 10 µM against a panel of 95 epigenetic targets. HDACs, DNMTs, and UTX IC50 values are indicated. C Predicted complex of CM-444 and CM-1758 with HDAC1, HDAC6, and HDAC7. D H3Ac and H3K27me3 levels were detected by western blot after daily treatment of an HL-60 cell line with 270 nM CM-444 or 300 nM CM-1758 for 48 h. H3 total was used as the loading control (representative experiment of 2 biologically independent studies). E DNA methylation of LINE-1 analyzed by pyrosequencing after daily treatment in HL-60 cell line with 270 nM CM-444 or 300 nM CM-1758 for 48 h. The DNA methylation percentage is indicated inside the circles. As a DNA methylated control, a universally methylated DNA was used. The data shown are the mean of two biologically independent experiments. F Dot blot was used to detect global 5-methylcytosine levels after CM-444 and CM-1758 daily treatment for 48 h in an HL-60 cell line (270 nM and 300 nM, respectively). Methylene blue staining was used as a loading control (representative experiment of 2 biologically independent studies). PDE5: phosphodiesterase-5; HDACs: histone deacetylases; DNMTs: DNA methyltransferases; SIRTs: sirtuins; HDMs: histone demethylases; HATs: histone acetyltransferases; HMTs: histone methyltransferases; 5mC: 5-methylcytosine; MB: methylene blue. Uncropped blots and source data are provided as a Source data file.
