Functional interrogation of HOXA9 regulome in MLLr leukemia via reporter-based CRISPR/Cas9 screen

eLife

Xujie Zhao¹  Beisi Xu¹  Shaela Wright¹  Taosheng Chen²  Xiang Chen²  Yang Zhang²  Lianzhong Zhao²  Chunliang Li³  Rui Lu⁴  Xinyue Zhou⁵  Ying Shao⁵  Jie An⁵  Hyeong-Min Lee⁶  Hao Zhang⁶  Yang Zhou⁶  Judith Hyle⁶ 

[1] Cancer Biology Program/Comprehensive Cancer Center, St. Jude Children’s Research Hospital, Memphis, United States;O’Neal Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, United States;Department of Computational Biology, St. Jude Children’s Research Hospital, Memphis, United States;Department of Pharmaceutical Sciences, St. Jude Children’s Research Hospital, Memphis, United States;Department of Tumor Cell Biology, St. Jude Children’s Research Hospital, Memphis, United States;Division of Hematology/Oncology, University of Alabama at Birmingham, Birmingham, United States;

关键词: knock-in;    CRISPR screen;    HOXA9;    transcriptional regulation;    mll-rearranged leukemia;   

DOI  :  10.7554/eLife.57858

来源: DOAJ

【 摘 要 】

Aberrant HOXA9 expression is a hallmark of most aggressive acute leukemias, notably those with KMT2A (MLL) gene rearrangements. HOXA9 overexpression not only predicts poor diagnosis and outcome but also plays a critical role in leukemia transformation and maintenance. However, our current understanding of HOXA9 regulation in leukemia is limited, hindering development of therapeutic strategies. Here, we generated the HOXA9-mCherry knock-in reporter cell lines to dissect HOXA9 regulation. By utilizing the reporter and CRISPR/Cas9 screens, we identified transcription factors controlling HOXA9 expression, including a novel regulator, USF2, whose depletion significantly down-regulated HOXA9 expression and impaired MLLr leukemia cell proliferation. Ectopic expression of Hoxa9 rescued impaired leukemia cell proliferation upon USF2 loss. Cut and Run analysis revealed the direct occupancy of USF2 at HOXA9 promoter in MLLr leukemia cells. Collectively, the HOXA9 reporter facilitated the functional interrogation of the HOXA9 regulome and has advanced our understanding of the molecular regulation network in HOXA9-driven leukemia.

【 授权许可】

Unknown