MicroRNA Profiling of Human-Induced Pluripotent Stem Cells

Department of Bioengineering, Stanford University School of Medicine, Stanford, CA 94305, USA.
Stem cells and development (Impact Factor: 3.73). 02/2009; 18(5):749-58. DOI: 10.1089/scd.2008.0247
Source: PubMed


MicroRNAs (miRNAs) are a newly discovered endogenous class of small noncoding RNAs that play important posttranscriptional regulatory roles by targeting mRNAs for cleavage or translational repression. Accumulating evidence now supports the importance of miRNAs for human embryonic stem cell (hESC) self-renewal, pluripotency, and differentiation. However, with respect to induced pluripotent stem cells (iPSC), in which embryonic-like cells are reprogrammed from adult cells using defined factors, the role of miRNAs during reprogramming has not been well-characterized. Determining the miRNAs that are associated with reprogramming should yield significant insight into the specific miRNA expression patterns that are required for pluripotency. To address this lack of knowledge, we use miRNA microarrays to compare the "microRNA-omes" of human iPSCs, hESCs, and fetal fibroblasts. We confirm the presence of a signature group of miRNAs that is up-regulated in both iPSCs and hESCs, such as the miR-302 and 17-92 clusters. We also highlight differences between the two pluripotent cell types, as in expression of the miR-371/372/373 cluster. In addition to histone modifications, promoter methylation, transcription factors, and other regulatory control elements, we believe these miRNA signatures of pluripotent cells likely represent another layer of regulatory control over cell fate decisions, and should prove important for the cellular reprogramming field.

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    • "More than 20 individuals or families of miRNAs play critical roles in self-renewal and pluripotency in ESCs and iPSCs by regulating numerous target genes (Moradi et al., 2014; Greve et al., 2013); however, their connections with LIF/Stat3 remains ill defined, and miR-205-5p is no exception. Like mouse miR-205-5p, human miR-205-5p, whose sequence is identical to its mouse counterpart, is more highly expressed in human ESCs and iPSCs than in fibroblasts (Wilson et al., 2009). In addition, Oct4, Sox2, and Nanog proteins bind to the genomic locus of miR-205 in ESCs, suggesting direct regulation by the pluripotency master proteins (Marson et al., 2008). "
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    ABSTRACT: Leukemia inhibitory factor (LIF) is widely used to establish and maintain naïve pluripotent stem cells, including mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). Although the combination of chemical inhibitors called 2i can establish mouse iPSCs without LIF from primed pluripotent stem cells, it has been difficult, if not impossible, to establish mouse iPSCs from differentiated somatic cells without LIF. We previously showed that the fusion gene of the transactivation domain of MyoD and the full-length Oct4 (M3O) increases the efficiency of making iPSCs when transduced into fibroblasts along with Sox2, Klf4, and c-Myc (M3O-SKM). Here, we report that M3O-SKM allows for establishment of iPSCs without exogenous LIF from mouse embryonic fibroblasts. The established iPSCs remained undifferentiated and maintained pluripotency over 90days without LIF as long as M3O was expressed. The iPSCs upregulated miR-205-5p, which was potentially involved in the LIF-independence by suppressing the two signaling pathways inhibited by 2i. The result indicates that potentiated Oct4 can substitute for the LIF signaling pathway, providing a novel model to link Oct4 and LIF, two of the most significant players in naïve pluripotency.
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    • "Although the functions of the majority of newly discovered non-coding RNAs are still unknown, some were found to play important roles in the regulation of stem cells. Recent studies concentrate on miRNAs (Wilson et al., 2009; Kim et al., 2011; Lipchina et al., 2011). In the context of stem cell biology, of particular interest is the role of these RNAs in expression of renewal genes in human embryonic stem cells (hESCs) or in regulation of induced pluripotency (Li et al., 2011). "
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    ABSTRACT: Several studies have demonstrated the important role of non-coding RNAs as regulators of posttranscriptional processes, including stem cells self-renewal and neural differentiation. Human embryonic stem cells (hESCs) and induced pluripotent stem cells (ihPSCs) show enormous potential in regenerative medicine due to their capacity to differentiate to virtually any type of cells of human body. Deciphering the role of non-coding RNAs in pluripotency, self-renewal and neural differentiation will reveal new molecular mechanisms involved in induction and maintenances of pluripotent state as well as triggering these cells toward clinically relevant cells for transplantation. In this brief review we will summarize recently published studies which reveal the role of non-coding RNAs in pluripotency and neural differentiation of hESCs and ihPSC.
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    • "Involvement of different miRNA clusters in the activation and inhibition of the specific cellular processes during reprogramming is presented on Figure 3. The mir-302 cluster is located in the 4q25 locus of human chromosome 4 (Puca et al. 2001) and is predominantly expressed in hES and iPS cells (Suh et al. 2004, Wilson et al. 2009), while during early embryonic development and in vitro differentiation the expression of miR-302 is lost (Suh et al. 2004, Ren et al. 2009). The majority of miR-302-targeted genes are transcripts of developmental signals and oncogenes (Lin et al. 2008). "
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    ABSTRACT: Derivation of pluripotent stem cells from adult somatic tissues by reprogramming technology has opened new therapeutic possibilities. Current most efficient procedures for derivation of induced pluripotent stem (iPS) cells are based on the viral vectors, which represent the danger of insertional mutagenesis during incorporation of introduced genes into the host genome. To circumvent this problem, the new, safe, non-integrative and non-viral strategies of reprogramming have been developed. In this review we discuss novel DNA-free and viral-free methods of reprogramming to iPS cells including protein transduction, mRNA and microRNA delivery.
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