The hsa-miR-302 cluster controls ectodermal differentiation of human pluripotent stem cell via repression of DAZAP2
The hsa-miR-302 cluster controls ectodermal differentiation of human pluripotent stem cell via repression of DAZAP2
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DOI：
10.1016/j.reth.2020.03.011
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发表时间：
2020-12-01
影响因子：
4.3
通讯作者：
Akutsu, Hidenori
中科院分区：
文献类型：
--
作者：
Sugawara, Tohru;Miura, Takumi;Akutsu, Hidenori
简介：最近的研究表明，microRNA（miRNAs，miRs）是重要的自我更新，分化，和细胞重编程的体细胞成诱导多能干细胞（iPSC）;然而，它们的功能作用和靶基因的人PSC特异性miRs，包括hsa-miR-302簇调控仍然在很大程度上是未知的。方法：我们分析了4个体细胞系、8个来源于4种不同细胞类型的人iPSC系、3个人ESC系和从人ESC分化的胚状体中miR的表达谱，以鉴定人PSC特异性miR。我们还分析了miR和mRNA的同时表达谱，以鉴定人PSC特异性miR的候选靶标。结果：筛选出22个hsa-miR-302簇的候选靶基因，这些靶基因在未分化的人PSC中中度表达，在分化的人PSC中表达上调。hsa-miR-302 a、-302b、-302c和-302d直接抑制了其中一个靶点-无精子症相关蛋白2（DAZAP 2），但hsa-miR-367不抑制DAZAP 2。DAZAP 2的过表达导致未分化的人iPSC的细胞增殖减少，尽管形态学和未分化标记基因表达不受影响。结论：hsa-miR-302簇通过抑制DAZAP 2来调控人PSC的细胞增殖和神经分化，从而突出了人PSC特异性miR在维持多能性方面的额外功能。(C)2020年，日本再生医学学会。制作和主办：Elsevier B. V.
Introduction: Recent studies have revealed that microRNAs (miRNAs, miRs) are important for self-renewal, differentiation, and cellular reprogramming of somatic cells into induced pluripotent stem cells (iPSC); however, their functional roles and target genes that are regulated by human PSC-specific miRs including hsa-miR-302 clusters remain largely unknown. Analysis of their target gene will give us the opportunity to understand the functional roles of such miRs.Methods: We analyzed the expression profiles of miRs in 4 somatic cell lines, 8 human iPSC lines derived from 4 different cell types, 3 human ESC lines, and embryoid bodies differentiated from the human ESCs to identify human PSC-specific miRs. We also analyzed the simultaneous expression profiles of miRs and mRNAs to identify candidate targets of human PSC-specific miRs. Then, we constructed a vector for overexpressing one of the target gene to dissect the functions of human PSC-specific miR in maintenance of self-renew and differentiation.Results: We focused on hsa-miR-302 cluster as a human PSC-specific miR and identified 22 candidate targets of hsa-miR-302 cluster that were moderately expressed in undifferentiated human PSCs and upregulated in differentiated cells. Deleted in azoospermia-associated protein 2 (DAZAP2), one such target, was directly repressed by hsa-miR-302a, -302b, -302c and -302d, but not by hsa-miR-367. Overexpression of DAZAP2 caused a decrease in cell proliferation of undifferentiated human iPSCs, although morphology and undifferentiated marker gene expression was not affected. In addition, neural differentiation was suppressed in DAZAP2-overexpressing human iPSCs.Conclusion: Our study revealed that hsa-miR-302 cluster controls the cell proliferation of human PSCs and the neural differentiation of human PSCs by repression of DAZAP2, thereby highlighting an additional function of human PSC-specific miRs in maintaining pluripotency. (C) 2020, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.