Functional interrogation of HOXA9 regulome in MLLr leukemia via reporter-based CRISPR/Cas9 screen.
Hao ZhangYang ZhangXinyue ZhouShaela WrightJudith HyleLianzhong ZhaoJie AnXujie ZhaoYing ShaoBei-Si XuHyeong-Min LeeTaosheng ChenYang ZhouXiang ChenRui LuChunliang LiPublished in: eLife (2020)
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.
Keyphrases
- long non coding rna
- crispr cas
- long noncoding rna
- poor prognosis
- acute myeloid leukemia
- cell proliferation
- bone marrow
- transcription factor
- genome editing
- binding protein
- dna methylation
- induced apoptosis
- genome wide
- oxidative stress
- hepatitis b virus
- single molecule
- cell cycle
- endoplasmic reticulum stress
- liver failure
- acute respiratory distress syndrome
- cell death
- copy number
- respiratory failure