The Nodal inhibitor Lefty is negatively modulated by the microRNA miR-302 in human embryonic stem cells
The Nodal inhibitor Lefty is negatively modulated by the microRNA miR-302 in human embryonic stem cells
复制标题
DOI：
10.1096/fj.10-172221
复制
发表时间：
2011-05-01
期刊：
影响因子：
4.8
通讯作者：
Menendez, Pablo
中科院分区：
文献类型：
--
作者：
Barroso-delJesus, Alicia;Lucena-Aguilar, Gema;Menendez, Pablo
microRNA（miRNAs）在人类胚胎干细胞（hESC）的早期发育和维持中具有重要作用。miRNA miR-302-367在hESC中特异性表达，并且其表达在分化时衰减。我们先前鉴定了编码人miR-302-367簇的基因的结构并表征了其启动子。启动子活性在hESC中进行了功能验证，为进一步研究这些miRNA分子如何适应赋予hESC“干性”特性的复杂分子网络开辟了新的途径。特异性miRNA-mRNA相互作用的生理作用在很大程度上仍然未知。在这里，我们研究了hESC中推定的miR-302-367 mRNA靶点，可能与ESC生物学相关。我们发现Nodal抑制剂Lefty 1和Lefty 2在hESCs中被miR-302转录后靶向。功能分析表明，miR-302 s负调节lefties的水平，并成为TGF β/Nodal途径的上游调节因子，通过Smad-2/3信号传导发挥作用。miR-302-367簇在hESC中的过表达导致早期hESC分化延迟，如通过ESC特异性转录因子的水平提高所测量的，与移植有表达miR-302-367的hESC的小鼠中更快的畸胎瘤形成以及伴随的胚层特化受损相关联，显示早期中胚层、内胚层和外胚层特异性标志物的水平稳健降低。这些发现表明，Lefty在hESC中受到miR-302的负调节，这在维持多能性和胚层特化之间的平衡方面起着重要作用。Barroso-delJesus，A.，Lucena-Aguilar，G.，桑切斯湖Ligero，G.，古铁雷斯-阿兰达岛Menendez，P. Nodal抑制剂Lefty在人类胚胎干细胞中受到microRNA miR-302的负调节。FASEB J.25，1497-1508（2011）。www.fasebj.org
MicroRNAs (miRNAs) have been shown to be important in early development and maintenance of human embryonic stem cells (hESCs). The miRNA miR-302-367 is specifically expressed in hESCs, and its expression decays on differentiation. We previously identified the structure of the gene coding for the human miR-302-367 cluster and characterized its promoter. The promoter activity was functionally validated in hESCs, opening up new avenues to further investigate how these miRNA molecules fit in the complex molecular network conferring "stemness" properties to hESCs. The physiological roles of specific miRNA-mRNA interactions remain largely unknown. Here, we investigated putative miR-302-367 mRNA targets in hESCs, potentially relevant for ESC biology. We found that the Nodal inhibitors Lefty1 and Lefty2 are post-transcriptionally targeted by miR-302s in hESCs. Functional analyses indicate that miR-302s negatively modulate the level of lefties, and become upstream regulators of the TGF beta/Nodal pathway, functioning via Smad-2/3 signaling. Overexpression of the miR-302-367 cluster in hESCs causes a delay in early hESC differentiation, as measured by enhanced levels of ESC-specific transcription factors, coupled to a faster teratoma formation in mice transplanted with miR-302-367-expressing hESCs and a concomitant impairment of germ layer specification, displaying robust decreased levels of early mesodermal, endodermal, and ectodermal specific markers. These findings suggest that Lefty is negatively modulated by miR-302s in hESCs, which plays an important role in maintaining the balance between pluripotency and germ layer specification.-Barroso-delJesus, A., Lucena-Aguilar, G., Sanchez, L., Ligero, G., Gutierrez-Aranda, I., Menendez, P. The Nodal inhibitor Lefty is negatively modulated by the microRNA miR-302 in human embryonic stem cells. FASEB J. 25, 1497-1508 (2011). www.fasebj.org