人类附睾中的转录网络。
Transcriptional networks in the human epididymis.
机构信息
Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH, 44106, USA.
出版信息
Andrology. 2019 Sep;7(5):741-747. doi: 10.1111/andr.12629. Epub 2019 May 2.
BACKGROUND
The epithelial lining of the human epididymis is critical for sperm maturation. This process requires distinct specialized functions in the head, body, and tail of the duct. These region-specific properties are maintained by distinct gene expression profiles which are governed by transcription factor networks, non-coding RNAs, and other factors.
MATERIALS AND METHODS
We used genome-wide protocols including DNase-seq, RNA-seq and ChIP-seq to characterize open (active) chromatin, the transcriptome and occupancy of specific transcription factors (TFs) respectively, in caput, corpus, and cauda segments of adult human epididymis tissue and primary human epididymis epithelial (HEE) cell cultures derived from them. RNA-seq following TF depletion or activation, combined with gene ontology analysis also determined TF targets.
RESULTS
Among regional differentially expressed transcripts were epithelial-selective transcription factors (TFs), microRNAs, and antiviral response genes. Caput-enriched TFs included hepatocyte nuclear factor 1 (HNF1) and the androgen receptor (AR), both of which were also predicted to occupy cis-regulatory elements identified as open chromatin in HEE cells. HNF1 targets were identified genome-wide using ChIP-seq, in HEE cells. Next, siRNA-mediated depletion of HNF1 revealed a pivotal role for this TF in coordinating epithelial water and solute transport in caput epithelium. The importance of AR in HEE cells was shown by AR ChIP-seq, and by RNA-seq after synthetic androgen (R1881) treatment. AR has a distinct transcriptional program in the HEE cells and likely recruits different co-factors (RUNX1 and CEBPβ) in comparison to those used in prostate epithelium.
DISCUSSION AND CONCLUSION
Our data identify many transcription factors that regulate the development and differentiation of HEE cells. Moreover, a comparison between immature and adult HEE cells showed key TFs in the transition to fully differentiated function of this epithelium. These data may help identify new targets to treat male infertility and have the potential to open new avenues for male contraception.
背景
人类附睾的上皮衬里对精子成熟至关重要。这个过程需要在管的头部、体部和尾部具有独特的专门功能。这些区域特异性特性是由转录因子网络、非编码 RNA 和其他因素控制的不同基因表达谱维持的。
材料和方法
我们使用全基因组方案，包括 DNase-seq、RNA-seq 和 ChIP-seq，分别描述了成人附睾组织和衍生自它们的原代人附睾上皮（HEE）细胞培养物的头部、体部和尾部的开放（活跃）染色质、转录组和特定转录因子（TF）的占有率。TF 耗竭或激活后的 RNA-seq 结合基因本体分析也确定了 TF 靶标。
结果
在区域差异表达的转录本中，包括上皮选择性转录因子（TFs）、microRNAs 和抗病毒反应基因。头部富集的 TF 包括肝细胞核因子 1（HNF1）和雄激素受体（AR），这两种 TF 都被预测为在 HEE 细胞中占据作为开放染色质的顺式调节元件。在 HEE 细胞中，使用 ChIP-seq 鉴定了 HNF1 靶标。接下来，siRNA 介导的 HNF1 耗竭揭示了该 TF 在协调头部上皮的水和溶质转运中的关键作用。AR 在 HEE 细胞中的重要性通过 AR ChIP-seq 和合成雄激素（R1881）处理后的 RNA-seq 显示出来。AR 在 HEE 细胞中有一个独特的转录程序，并且可能在与前列腺上皮相比时招募不同的共因子（RUNX1 和 CEBPβ）。
讨论和结论
我们的数据确定了许多调节 HEE 细胞发育和分化的转录因子。此外，对不成熟和成年 HEE 细胞的比较显示了该上皮向完全分化功能转变的关键 TF。这些数据可能有助于确定治疗男性不育的新靶点，并有可能为男性避孕开辟新途径。
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