PDF(Pubmed)
Abstract：
Temporal regulation of super-enhancer (SE) driven transcription factors (TFs) underlies normal developmental programs. Neuroblastoma (NB) arises from an inability of sympathoadrenal progenitors to exit a self-renewal program and terminally differentiate. To identify SEs driving TF regulators, we use all-trans retinoic acid (ATRA) to induce NB growth arrest and differentiation. Time-course H3K27ac ChIP-seq and RNA-seq reveal ATRA coordinated SE waves. SEs that decrease with ATRA link to stem cell development (MYCN, GATA3, SOX11). CRISPR-Cas9 and siRNA verify SOX11 dependency, in vitro and in vivo. Silencing the SOX11 SE using dCAS9-KRAB decreases SOX11 mRNA and inhibits cell growth. Other TFs activate in sequential waves at 2, 4 and 8 days of ATRA treatment that regulate neural development (GATA2 and SOX4). Silencing the gained SOX4 SE using dCAS9-KRAB decreases SOX4 expression and attenuates ATRA-induced differentiation genes. Our study identifies oncogenic lineage drivers of NB self-renewal and TFs critical for implementing a differentiation program.
摘要：
超增强子（SE）驱动的转录因子（TF）的时间调节是正常发育程序的基础。神经母细胞瘤（NB）是由于交感肾上腺祖细胞无法退出自我更新程序并最终分化而引起的。为了识别驱动TF调节器的SE，我们使用全反式维甲酸（ATRA）诱导NB生长停滞和分化。时程H3K27acChIP-seq和RNA-seq揭示了ATRA协调的SE波。与ATRA一起减少的SE与干细胞发育有关(MYCN，GATA3，SOX11）。CRISPR-Cas9和siRNA验证SOX11依赖性，在体外和体内。使用dCAS9-KRAB沉默SOX11SE可降低SOX11mRNA并抑制细胞生长。在ATRA治疗的第2、4和8天时，其他TF在调节神经发育的连续波中激活（GATA2和SOX4）。使用dCAS9-KRAB沉默获得的SOX4SE会降低SOX4表达并减弱ATRA诱导的分化基因。我们的研究确定了NB自我更新的致癌谱系驱动因素和对实施分化计划至关重要的TFs。
