Xeno-sensing activity of the aryl hydrocarbon receptor in human pluripotent stem cell-derived hepatocyte-like cells.
Xeno-sensing activity of the aryl hydrocarbon receptor in human pluripotent stem cell-derived hepatocyte-like cells.
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DOI：
10.1038/srep21684
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发表时间：
2016-02-22
影响因子：
4.6
通讯作者：
Park HJ
中科院分区：
文献类型：
--
作者：
Kim HM;Kim JW;Choi Y;Chun HS;Im I;Han YM;Song CW;Yoon S;Park HJ
虽然来源于人多能干细胞的肝细胞样细胞（hpsc - hlc）被认为是预测肝毒性的一种有前途的模型，但由于药物代谢酶（DMEs）活性低，其应用受到限制。在这里，我们发现外源受体（组成型雄甾烷受体，CAR和妊娠X受体，PXR）的低表达导致了hPSC-HLCs中DMEs的低活性。大多数CAR-和pxr调控的DMEs和转运体在hpsc - hplc中转录下调。在hPSC-HLCs中，CAR和PXR的转录表达被高度抑制，而芳烃受体（AHR）的mRNA水平与成人肝脏相当。此外，在hPSC-HLCs中，仅在AHR处观察到配体诱导的转录激活。亚硫酸氢盐测序分析表明，在hPSC-HLCs中，启动子CAR和PXR的超甲基化与转录活性降低有关。ahr选择性配体通过AHR-DNA在异种反应元件上的直接结合，增加了ahr依赖性靶基因的转录。此外，AHR拮抗剂显著抑制AHR依赖性靶基因的表达。因此，AHR在hpsc - hlc中可以作为异种传感器和配体依赖性转录因子。我们的研究结果表明，hPSC-HLCs可用于筛选与AHR信号相关的有毒物质，并确定潜在的AHR靶向治疗方法。
Although hepatocyte-like cells derived from human pluripotent stem cells (hPSC-HLCs) are considered a promising model for predicting hepatotoxicity, their application has been restricted because of the low activity of drug metabolizing enzymes (DMEs). Here we found that the low expression of xenobiotic receptors (constitutive androstane receptor, CAR; and pregnane X receptor, PXR) contributes to the low activity of DMEs in hPSC-HLCs. Most CAR- and PXR-regulated DMEs and transporters were transcriptionally down-regulated in hPSC-HLC. Transcriptional expression of CAR and PXR was highly repressed in hPSC-HLCs, whereas mRNA levels of aryl hydrocarbon receptor (AHR) were comparable to those of adult liver. Furthermore, ligand-induced transcriptional activation was observed only at AHR in hPSC-HLCs. Bisulfite sequencing analysis demonstrated that promoter hypermethylation of CAR and PXR was associated with diminished transcriptional activity in hPSC-HLCs. Treatment with AHR-selective ligands increased the transcription of AHR-dependent target genes by direct AHR-DNA binding at the xenobiotic response element. In addition, an antagonist of AHR significantly inhibited AHR-dependent target gene expression. Thus, AHR may function intrinsically as a xenosensor as well as a ligand-dependent transcription factor in hPSC-HLCs. Our results indicate that hPSC-HLCs can be used to screen toxic substances related to AHR signaling and to identify potential AHR-targeted therapeutics.