RNA-seq analysis of human KMT2A rearranged MV4;11 AML cell line treated with DOT1L and/or EZH2 inhibitor

The histone methyltransferases DOT1L (H3K79me1,2,3 KMT, "activating" chromatin mark) and EZH2 (H3K27me1,2,3 KMT, "silencing" mark) have both been shown to be required for growth and survival of KMT2A rearranged AML cells. Both KMTs have been shown to modulate expression of HOXA cluster genes, albeit for EZH2 this has not been shown in the context of KMT2A rearranged AML, but in other subtypes of AML. We asked what transcriptional effects dual inhibition of DOT1L and EZH2 has in KMT2A rearranged AML. MV4;11 AML cells were exposed to the DOT1L inhibitor EPZ5676 at 0.08 and 0.16 uM, and the EZH2 inhibitor GSK126 at 0.3 uM for 7 days. Fresh media and compound were supplied on day 4. RNA was isolated using RNeasy (Quiagen) and submitted for RNA-Seq

组蛋白甲基转移酶（histone methyltransferases）DOT1L（介导H3K79me1/2/3的赖氨酸甲基转移酶（lysine methyltransferase, KMT），属于“活化型”染色质修饰标记）与EZH2（介导H3K27me1/2/3的赖氨酸甲基转移酶（KMT），属于“沉默型”染色质修饰标记），均被证实为KMT2A重排急性髓系白血病（acute myeloid leukemia, AML）细胞的生长与存活所必需。两类赖氨酸甲基转移酶均被发现可调控HOXA簇（HOXA cluster）基因的表达；不过针对EZH2的该调控作用尚未在KMT2A重排AML的背景中得到验证，仅在其他AML亚型中已有相关报道。本研究旨在探究DOT1L与EZH2的双重抑制对KMT2A重排AML细胞所产生的转录调控效应。将MV4;11 AML细胞分别用浓度为0.08 μM与0.16 μM的DOT1L抑制剂EPZ5676，以及0.3 μM的EZH2抑制剂GSK126处理7天；于第4天更换新鲜培养基并补加受试化合物。采用RNeasy试剂盒（Qiagen）提取总RNA，并将样品提交进行RNA测序（RNA-Seq）。