CRISPR screen of venetoclax response-associated genes identifies transcription factor ZNF740 as a key functional regulator.

IF 8.1 1区 生物学 Q1 CELL BIOLOGY
Lixia Zhang, Xinyue Zhou, Sajesan Aryal, Virginia Veasey, Pengcheng Zhang, Fu Jun Li, Yu Luan, Ravi Bhatia, Yang Zhou, Rui Lu
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Abstract

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.

Abstract Image

对 Venetoclax 反应相关基因进行 CRISPR 筛选,发现转录因子 ZNF740 是一个关键的功能调节因子。
BCL-2抑制剂(如venetoclax)为急性髓性白血病(AML)和其他癌症带来了治疗希望,但耐药性构成了重大挑战。了解调控 venetoclax 反应的机制至关重要。虽然相关研究发现了许多与 Venetoclax 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。在本研究中,我们针对对 venetoclax 敏感的原发性 AML 样本中约 1400 个上调基因进行了 CRISPR 基因敲除筛选,以评估它们对 venetoclax 反应的直接影响。我们的筛选发现转录因子ZNF740是一个关键的调控因子,它的表达可持续预测FAB分类中不同亚型的Venetoclax敏感性。ZNF740缺失会导致对文曲康的耐药性增加,而其过度表达则会提高对该药物的敏感性。从机理上讲,我们的转录组和基因组综合分析确定了NOXA是ZNF740的直接靶标,它负向调节MCL-1蛋白的稳定性。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缺失的细胞中恢复NOXA的表达可使AML细胞对venetoclax治疗重新敏感。此外,我们还证明,MCL-1和BCL-2的双重靶向可有效治疗体内ZNF740缺陷型AML。总之,我们的工作系统地阐明了 Venetoclax 反应特征基因之间的因果关系,并将 ZNF740 确立为调控 Venetoclax 敏感性的新型转录因子。
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来源期刊
Cell Death & Disease
Cell Death & Disease CELL BIOLOGY-
CiteScore
15.10
自引率
2.20%
发文量
935
审稿时长
2 months
期刊介绍: Brought to readers by the editorial team of Cell Death & Differentiation, Cell Death & Disease is an online peer-reviewed journal specializing in translational cell death research. It covers a wide range of topics in experimental and internal medicine, including cancer, immunity, neuroscience, and now cancer metabolism. Cell Death & Disease seeks to encompass the breadth of translational implications of cell death, and topics of particular concentration will include, but are not limited to, the following: Experimental medicine Cancer Immunity Internal medicine Neuroscience Cancer metabolism
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