Multiplex gene editing models of del(7q) reveal combined CUX1 and EZH2 loss drives clonal expansion and drug resistance

Blood Neoplasia Pub Date : 2025-03-03 DOI:10.1016/j.bneo.2025.100083

Matthew R. M. Jotte , Angela Stoddart , Tanner C. Martinez , Raven Moten , Yuqing Xue , Molly K. Imgruet , Hunter Blaylock , Henna S. Nam , Bonnie Hu , Jermaine Austin , Ningfei An , Saira Khan , Sandeep K. Gurbuxani , Megan E. McNerney

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Abstract

Loss of all or part of chromosome 7 [−7/del(7q)] is recurrent in myeloid neoplasms and associated with a poor response to chemotherapy. Chromosome 7–encoded genes driving drug resistance and the consequences of combinatorial 7q tumor suppressor gene loss have remained unclear, the latter question largely because of the challenges of modeling aneuploidy. Here, we use in silico data mining to uncover 7q genes involved in chemotherapy resistance. We establish murine models of del(7q) clonal hematopoiesis and drug resistance with multiplex CRISPR-Cas9 (CRISPR–associated protein 9)–mediated inactivation of 4 genes, Cux1, Ezh2, Kmt2c, and Kmt2e. Postgenotoxic exposure, combined deficiency of Cux1 and Ezh2 preferentially promotes clonal myeloid expansion in vivo, with compounding defects in DNA damage recognition and repair. Human acute myeloid leukemia cell lines similarly illustrate central roles for CUX1 and EZH2 loss in survival and DNA damage resolution after chemotherapy exposure. Transcriptome analysis reveals combined Cux1 and Ezh2 loss recapitulates gene signatures of -7 patients and defective DNA damage response pathways, to a greater extent than single gene loss. This work reveals a genetic interaction between CUX1 and EZH2, and sheds light on how −7/del(7q) contributes to leukemogenesis and drug resistance characteristic of these adverse-risk neoplasms. These data support the concept of 7q as a contiguous gene syndrome region, in which combined loss of multiple gene drives pathogenesis. Furthermore, our CRISPR-based approach may serve as a framework for interrogating other recurrent aneuploid events in cancer.

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del(7q)的多重基因编辑模型显示，CUX1和EZH2的联合缺失会导致克隆扩增和耐药性

7号染色体的全部或部分缺失[−7/del(7q)]在髓系肿瘤中是复发性的，并且与化疗反应差有关。7号染色体编码基因驱动耐药性和组合7q肿瘤抑制基因丢失的后果仍然不清楚，后者的问题主要是因为建模非整倍体的挑战。在这里，我们使用计算机数据挖掘来揭示与化疗耐药有关的7q基因。我们通过多重CRISPR-Cas9 （CRISPR-associated protein 9）介导的Cux1、Ezh2、Kmt2c和Kmt2e基因失活，建立小鼠del（7q）克隆造血和耐药模型。基因中毒暴露后，Cux1和Ezh2的联合缺乏在体内优先促进克隆性髓细胞的扩增，并导致DNA损伤识别和修复的复合缺陷。人类急性髓性白血病细胞系同样表明CUX1和EZH2缺失在化疗暴露后的生存和DNA损伤修复中起着核心作用。转录组分析显示，Cux1和Ezh2的联合缺失再现了-7例患者的基因特征和DNA损伤反应途径的缺陷，其程度大于单个基因缺失。这项工作揭示了CUX1和EZH2之间的遗传相互作用，并揭示了−7/del（7q）如何促进这些不良风险肿瘤的白血病发生和耐药特征。这些数据支持7q作为一个连续基因综合征区域的概念，其中多个基因的联合丢失驱动发病机制。此外，我们基于crispr的方法可以作为询问癌症中其他复发性非整倍体事件的框架。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Blood Neoplasia

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