具有平衡染色体易位的急性髓系白血病 (AML) 中的异常基因表达模式通常与表观遗传修饰因子的失调有关。由 (8;21)(q22;q22) 易位引起的 AML1/ETO (RUNX1/MTG8) 融合蛋白会导致其靶基因的表观遗传抑制。我们的目标是通过 shRNA 文库筛选和全局转录组学方法来确定 AML1/ETO 阳性 AML 细胞增殖和生存所依赖的关键表观遗传修饰因子。通过 shRNA 文库筛选，我们在两种 AML1/ETO 阳性细胞系 Kasumi-1 和 SKNO-1 中鉴定出了 41 个常见缺失基因。我们使用几种 AML1/ETO 阳性和阴性细胞系从遗传学和药理学角度验证了 DNMT1 和 ATR。我们还证明了 AML1/ETO 阳性 AML 患者接受 DNMT1 抑制剂地西他滨治疗后，成髓细胞的体内分化。对 9/14/18-U937 细胞中 AML1/ETO 诱导后的整体转录组学进行生物信息分析，鉴定出 973 个差异表达基因 (DEG)。三个基因（PARP2、PRKCD 和 SMARCA4）在 AML1/ETO 诱导后均下调，并在 shRNA 筛选中得到鉴定。总之，使用无偏的 shRNA 文库筛选和全局转录组学，我们已经确定了 AML1/ETO 阳性 AML 增殖的几个驱动表观遗传调节因子。 DNMT1 和 ATR 已经过验证，并且易于受到小分子的药理学抑制，显示出有希望的临床前和临床疗效。© 2024 作者。约翰·威利出版的《国际癌症杂志》

Aberrant gene expression patterns in acute myeloid leukemia (AML) with balanced chromosomal translocations are often associated with dysregulation of epigenetic modifiers. The AML1/ETO (RUNX1/MTG8) fusion protein, caused by the translocation (8;21)(q22;q22), leads to the epigenetic repression of its target genes. We aimed in this work to identify critical epigenetic modifiers, on which AML1/ETO-positive AML cells depend on for proliferation and survival using shRNA library screens and global transcriptomics approaches. Using shRNA library screens, we identified 41 commonly depleted genes in two AML1/ETO-positive cell lines Kasumi-1 and SKNO-1. We validated, genetically and pharmacologically, DNMT1 and ATR using several AML1/ETO-positive and negative cell lines. We also demonstrated in vivo differentiation of myeloblasts after treatment with the DNMT1 inhibitor decitabine in a patient with an AML1/ETO-positive AML. Bioinformatic analysis of global transcriptomics after AML1/ETO induction in 9/14/18-U937 cells identified 973 differentially expressed genes (DEGs). Three genes (PARP2, PRKCD, and SMARCA4) were both downregulated after AML1/ETO induction, and identified in shRNA screens. In conclusion, using unbiased shRNA library screens and global transcriptomics, we have identified several driver epigenetic regulators for proliferation in AML1/ETO-positive AML. DNMT1 and ATR were validated and are susceptible to pharmacological inhibition by small molecules showing promising preclinical and clinical efficacy.© 2024 The Author(s). International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.