Venetoclax 等 BCL-2 抑制剂为急性髓性白血病 (AML) 和其他癌症提供了治疗前景，但耐药性带来了重大挑战。了解调节 Venetoclax 反应的机制至关重要。虽然相关研究已经确定了许多与维奈托克敏感性相关的基因，但它们对药物反应的直接影响仍不清楚。在这项研究中，我们针对 Venetoclax 敏感的原发性 AML 样本中上调的约 1400 个基因进行了 CRISPR 敲除筛选，以评估它们对 Venetoclax 反应的直接影响。我们的筛选将转录因子 ZNF740 确定为关键调节因子，其表达一致地预测 FAB 分类亚型中的维奈托克敏感性。 ZNF740 耗竭会导致对 Ventoclax 的耐药性增加，而其过度表达会增强对该药物的敏感性。从机制上讲，我们的综合转录组学和基因组分析确定 NOXA 是 ZNF740 的直接靶标，ZNF740 负向调节 MCL-1 蛋白稳定性。 ZNF740 的缺失会下调 NOXA 并增加 AML 细胞中 MCL-1 的稳态蛋白水平。在 ZNF740 耗尽的细胞中恢复 NOXA 表达可使 AML 细胞对 Venetoclax 治疗重新敏感。此外，我们证明了 MCL-1 和 BCL-2 的双重靶向可有效治疗体内 ZNF740 缺陷的 AML。我们的工作共同系统地阐明了 Venetoclax 反应特征基因之间的因果关系，并将 ZNF740 确立为调节 Venetoclax 敏感性的新型转录因子。© 2024。作者。

BCL-2 inhibitors such as venetoclax offer therapeutic promise in acute myeloid leukemia (AML) and other cancers, but drug resistance poses a significant challenge. It is crucial to understand the mechanisms that regulate venetoclax response. While correlative studies have identified numerous genes linked to venetoclax sensitivity, their direct impact on the drug response remains unclear. In this study, we targeted around 1400 genes upregulated in venetoclax-sensitive primary AML samples and carried out a CRISPR knockout screen to evaluate their direct effects on venetoclax response. Our screen identified the transcription factor ZNF740 as a critical regulator, with its expression consistently predicting venetoclax sensitivity across subtypes of the FAB classification. ZNF740 depletion leads to increased resistance to ventoclax, while its overexpression enhances sensitivity to the drug. Mechanistically, our integrative transcriptomic and genomic analysis identifies NOXA as a direct target of ZNF740, which negatively regulates MCL-1 protein stability. Loss of ZNF740 downregulates NOXA and increases the steady state protein levels of MCL-1 in AML cells. Restoring NOXA expression in ZNF740-depleted cells re-sensitizes AML cells to venetoclax treatment. Furthermore, we demonstrated that dual targeting of MCL-1 and BCL-2 effectively treats ZNF740-deficient AML in vivo. Together, our work systematically elucidates the causal relationship between venetoclax response signature genes and establishes ZNF740 as a novel transcription factor regulating venetoclax sensitivity.© 2024. The Author(s).