Redox-Sensitive Transcription Factor NRF2 Enhances Trophoblast Differentiation via Induction of miR-1246 and Aromatase
Redox-Sensitive Transcription Factor NRF2 Enhances Trophoblast Differentiation via Induction of miR-1246 and Aromatase
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DOI：
10.1210/en.2017-03024
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发表时间：
2018-05-01
期刊：
影响因子：
4.8
通讯作者：
Mendelson, Carole R.
中科院分区：
文献类型：
--
作者：
Muralimanoharan, Sribalasubashini;Kwak, Youn-Tae;Mendelson, Carole R.
胎盘缺氧导致滋养层细胞侵袭和分化的失调可导致妊娠高血压疾病--子痫前期。在此，我们研究了miR-1246在人合体滋养层细胞分化过程中的作用和调控。MIR-1246针对WNT/β-连环蛋白信号转导的抑制剂GSK3b和AXIN2，该信号对胎盘发育至关重要，预计将针对JARID2，后者促进发育调节基因的沉默。在20%O-2中培养的人细胞滋养层细胞在hCyP191a/芳香酶的诱导下自发分化为合体滋养层细胞，这是分化的标志。在合体滋养层细胞分化过程中，在20%O-2中，MIR-1246被诱导150倍，而靶标GSK3b、AXIN2和JARID2显著减少。然而，当细胞滋养层细胞在2%O-2中培养时，miR-1246和芳香酶的诱导被抑制。重度子痫前期患者胎盘组织中MIR-1246显著降低，AXIN2、GSK3b和JARID2显著升高。为了确定调控miR-1246的因素，我们研究了氧化还原调节的转录因子NRF2，它预测了miR-1246启动子中的结合位点。有趣的是，NRF2信使RNA在合体滋养层细胞分化过程中上调，并在缺氧和先兆子痫胎盘中显著降低。此外，细胞滋养层细胞中的NRF2基因敲除抑制了miR-1246和hCYP19A1的诱导，以及与胎盘分化有关的转录因子C/EBPβ和PPARγ的诱导。应用染色质免疫沉淀-定量聚合酶链式反应，我们发现在合体滋养层细胞分化过程中，内源性NRF2与miR-1246和hCYP191A启动子的结合增加。因此，NRF2通过诱导C/EBPB、PPARg、hCYP19A1和miR-1246促进合体滋养层细胞的分化，miR-1246针对WNT抑制剂和JARID2，在先兆子痫中调节失调。
Dysregulation of human trophoblast invasion and differentiation with placental hypoxia can result in preeclampsia, a hypertensive disorder of pregnancy. Herein, we characterized the role and regulation of miR-1246, which is markedly induced during human syncytiotrophoblast differentiation. miR-1246 targets GSK3b and AXIN2, inhibitors of WNT/beta-catenin signaling, which is crucial for placental development, and is predicted to target JARID2, which promotes silencing of developmentally regulated genes. Human cytotrophoblasts cultured in 20% O-2 spontaneously differentiate to syncytiotrophoblast with induction of hCYP191A/aromatase, a marker of differentiation. miR-1246 was induced > 150-fold during syncytiotrophoblast differentiation in 20% O-2, whereas targets-GSK3b, AXIN2, and JARID2-were significantly decreased. However, when cytotrophoblasts were cultured in 2% O-2, miR-1246 and aromatase induction were prevented. miR-1246 was significantly decreased in placentas of women with severe preeclampsia, whereas AXIN2, GSK3b, and JARID2 were increased, compared with normotensive subjects. To identify factors that regulate miR-1246, we investigated the redox-regulated transcription factor NRF2, which has predicted binding sites in the miR-1246 promoter. Intriguingly, NRF2 messenger RNA was upregulated during syncytiotrophoblast differentiation and significantly reduced by hypoxia and in preeclamptic placentas. Moreover, NRF2 knockdown in cytotrophoblasts inhibited induction of miR-1246 and hCYP19A1, as well as transcription factors C/EBP beta and PPAR gamma, which are implicated in placental differentiation. Using chromatin immunoprecipitation-quantitative polymerase chain reaction, we found that binding of endogenous NRF2 to the miR-1246 and hCYP191A promoters increased during syncytiotrophoblast differentiation. Thus, NRF2 promotes syncytiotrophoblast differentiation by inducing C/EBPb, PPARg, hCYP19A1, and miR-1246, which targets WNT inhibitors and JARID2 and is dysregulated in preeclampsia.