The Transcriptional Network That Controls Growth Arrest and Macrophage Differentiation in the Human Myeloid Leukemia Cell Line THP-1
The Transcriptional Network That Controls Growth Arrest and Macrophage Differentiation in the Human Myeloid Leukemia Cell Line THP-1
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DOI：
10.3389/fcell.2020.00498
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发表时间：
2020-07-03
影响因子：
5.5
通讯作者：
Summers, Kim M.
中科院分区：
文献类型：
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作者：
Gazova, Iveta;Lefevre, Lucas;Summers, Kim M.
人类急性髓系白血病细胞系THP-1对佛波酯的反应已被广泛研究，以测试候选白血病治疗方法，并作为细胞周期停滞和单核-巨噬细胞分化的模型。在这里，我们使用基因表达的帽分析(CAGE)分析了佛波酯(PMA)处理96h后THP-1细胞转录调控的密集时间过程。PMA处理显著减少了进入S期的细胞数量，并阻止了细胞退出G2/M。随着时间的推移，PMA处理的细胞变得黏附，成熟的巨噬细胞特异性基因的表达逐渐增加。在1-2小时内，PMA诱导肿瘤蛋白P53(TP53)的已知靶点，显著的CDKN1A，随后逐渐下调细胞周期相关基因。此外，在最初的2 h内，PMA还诱导了包括编码巨噬细胞分化相关蛋白的转录因子基因(EGR2、Jun、MAFB)和下调参与未成熟髓系细胞增殖的转录因子基因(MYB、IRF8、GFI1)的即刻早期基因。密集的时间进程显示对PMA的反应不是线性的和渐进性的。相反，基于网络的时间进程数据聚类突出了编码反馈调节因子的基因以及与巨噬细胞分化相关的转录因子及其推断的目标基因的瞬时上调和下调表达的顺序级联。CAGE还发现了在THP-1细胞中表达的已知和候选的新型增强子，以及许多目前缺乏功能注释和/或在巨噬细胞中没有已知功能的新型可诱导基因。时间进程可以在ZENBU平台上获得，可以与FANTOM4和FANTOM5数据进行比较。
The response of the human acute myeloid leukemia cell line THP-1 to phorbol esters has been widely studied to test candidate leukemia therapies and as a model of cell cycle arrest and monocyte-macrophage differentiation. Here we have employed Cap Analysis of Gene Expression (CAGE) to analyze a dense time course of transcriptional regulation in THP-1 cells treated with phorbol myristate acetate (PMA) over 96 h. PMA treatment greatly reduced the numbers of cells entering S phase and also blocked cells exiting G2/M. The PMA-treated cells became adherent and expression of mature macrophage-specific genes increased progressively over the duration of the time course. Within 1-2 h PMA induced known targets of tumor protein p53 (TP53), notablyCDKN1A, followed by gradual down-regulation of cell-cycle associated genes. Also within the first 2 h, PMA induced immediate early genes including transcription factor genes encoding proteins implicated in macrophage differentiation (EGR2, JUN, MAFB) and down-regulated genes for transcription factors involved in immature myeloid cell proliferation (MYB, IRF8, GFI1). The dense time course revealed that the response to PMA was not linear and progressive. Rather, network-based clustering of the time course data highlighted a sequential cascade of transient up- and down-regulated expression of genes encoding feedback regulators, as well as transcription factors associated with macrophage differentiation and their inferred target genes. CAGE also identified known and candidate novel enhancers expressed in THP-1 cells and many novel inducible genes that currently lack functional annotation and/or had no previously known function in macrophages. The time course is available on the ZENBU platform allowing comparison to FANTOM4 and FANTOM5 data.