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• W2061457029 abstract "Exploiting the properties of stem cells by microRNA (miRNA) profiling offers an attractive approach to identify new regulators of stem cell fate. Although numerous miRNA have been screened from hematopoietic stem cells (HSC), the targets corresponding to many of these miRNA have not yet been fully elucidated. By miRNA profiling in a subpopulation of CD34+ cells isolated from peripheral blood, we have identified eight clusters of miRNA that were differentially expressed. Further analysis of one of the clusters by bioinformatics revealed that a miRNA, miR-181a*, which is highly expressed in the adherent CD34+ cells, affects the expression levels of Nanog, a stem cell surrogate marker. We show specifically by reporter assay and mutational analysis that miR-181a* targets a seedless 3′ compensatory site in the 3′UTR of Nanog and affects gene expression. We demonstrate that inhibiting miR-181a* upregulates the Nanog expression level, in addition to an increase in alkaline phosphatase activity. Our studies suggest that miR-181a* may be important in controlling the expression level of Nanog in a subpopulation of CD34+ cells. Exploiting the properties of stem cells by microRNA (miRNA) profiling offers an attractive approach to identify new regulators of stem cell fate. Although numerous miRNA have been screened from hematopoietic stem cells (HSC), the targets corresponding to many of these miRNA have not yet been fully elucidated. By miRNA profiling in a subpopulation of CD34+ cells isolated from peripheral blood, we have identified eight clusters of miRNA that were differentially expressed. Further analysis of one of the clusters by bioinformatics revealed that a miRNA, miR-181a*, which is highly expressed in the adherent CD34+ cells, affects the expression levels of Nanog, a stem cell surrogate marker. We show specifically by reporter assay and mutational analysis that miR-181a* targets a seedless 3′ compensatory site in the 3′UTR of Nanog and affects gene expression. We demonstrate that inhibiting miR-181a* upregulates the Nanog expression level, in addition to an increase in alkaline phosphatase activity. Our studies suggest that miR-181a* may be important in controlling the expression level of Nanog in a subpopulation of CD34+ cells. In recent years, studies in embryonic and somatic stem cells have provided basic insight into the molecular and cellular properties of stem cells.1Moore KA Lemischka IR Stem cells and their niches.Science. 2006; 311: 1880-1885Crossref PubMed Scopus (1263) Google Scholar It is recognized that pluripotent stem cells such as embryonic stem cells (ESC) can replenish differentiated cell types and achieve long-term tissue reconstitution. Similar to ESC, adult stem cells also have the capacity for self-renewal and multilineage differentiation.2Weissman IL Stem cells: units of development, units of regeneration, and units in evolution.Cell. 2000; 100: 157-168Abstract Full Text Full Text PDF PubMed Scopus (1446) Google Scholar Adult stem cells constitute ~1–2% of the total cell population within a specific tissue and are essential for maintaining homeostasis. These cells are normally quiescent and are held in an undifferentiated state within their niche until they receive a signal to self-renew or differentiate.3Fuchs E Tumbar T Guasch G Socializing with the neighbors: stem cells and their niche.Cell. 2004; 116: 769-778Abstract Full Text Full Text PDF PubMed Scopus (1459) Google Scholar They reside in various tissue types including the bone marrow, brain, digestive system, skin, retina, muscles, pancreas, and liver.4Slack JM Stem cells in epithelial tissues.Science. 2000; 287: 1431-1433Crossref PubMed Scopus (314) Google Scholar Hematopoietic stem cells (HSC) are the most widely studied and characterized adult stem cells.5Orkin SH Zon LI Hematopoiesis: an evolving paradigm for stem cell biology.Cell. 2008; 132: 631-644Abstract Full Text Full Text PDF PubMed Scopus (1616) Google Scholar They play an essential role in sustaining the formation of the blood and the immune system.6Till JE McCulloch EA A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. 1961.Radiat Res. 2011; 175: 145-149Crossref PubMed Scopus (14) Google Scholar,7Kondo M Wagers AJ Manz MG Prohaska SS Scherer DC Beilhack GF et al.Biology of hematopoietic stem cells and progenitors: implications for clinical application.Annu Rev Immunol. 2003; 21: 759-806Crossref PubMed Scopus (769) Google Scholar These adult stem cells reside in the bone marrow along with mesenchymal stem cells (MSC).5Orkin SH Zon LI Hematopoiesis: an evolving paradigm for stem cell biology.Cell. 2008; 132: 631-644Abstract Full Text Full Text PDF PubMed Scopus (1616) Google Scholar,8Bianco P Riminucci M Gronthos S Robey PG Bone marrow stromal stem cells: nature, biology, and potential applications.Stem Cells. 2001; 19: 180-192Crossref PubMed Scopus (1710) Google Scholar It was previously thought that adult stem cells were lineage restricted; however, recent studies have shown that HSC and mesenchymal stem cells have enormous plasticity.9Kuçi S Kuçi Z Latifi-Pupovci H Niethammer D Handgretinger R Schumm M et al.Adult stem cells as an alternative source of multipotential (pluripotential) cells in regenerative medicine.Curr Stem Cell Res Ther. 2009; 4: 107-117Crossref PubMed Scopus (52) Google Scholar,10Pittenger MF Mackay AM Beck SC Jaiswal RK Douglas R Mosca JD et al.Multilineage potential of adult human mesenchymal stem cells.Science. 1999; 284: 143-147Crossref PubMed Scopus (17647) Google Scholar These characteristics have made them attractive for developing stem cell-based therapies. Moreover, they offer several advantages including ease of manipulation, lack of serious ethical issues and, in the autologous setting, the absence of immunogenicity. MicroRNAs (miRNAs) are short single-stranded RNA molecules of about 17–25 nucleotides that control gene expression post-transcriptionally in many different cellular processes either by blocking translation or by inducing mRNA degradation through sequence complementation.11Bartel DP MicroRNAs: target recognition and regulatory functions.Cell. 2009; 136: 215-233Abstract Full Text Full Text PDF PubMed Scopus (15352) Google Scholar,12Castanotto D Rossi JJ The promises and pitfalls of RNA-interference-based therapeutics.Nature. 2009; 457: 426-433Crossref PubMed Scopus (1020) Google Scholar,13Gangaraju VK Lin H MicroRNAs: key regulators of stem cells.Nat Rev Mol Cell Biol. 2009; 10: 116-125Crossref PubMed Scopus (580) Google Scholar,14He L Hannon GJ MicroRNAs: small RNAs with a big role in gene regulation.Nat Rev Genet. 2004; 5: 522-531Crossref PubMed Scopus (5497) Google Scholar Although numerous signaling pathways, transcription factors and epigenetic changes are essential cellular regulators in determining stem cell fate, recent studies have shown that miRNAs are also implicated in coordinating the necessary changes in gene expression.15Ivey KN Srivastava D MicroRNAs as regulators of differentiation and cell fate decisions.Cell Stem Cell. 2010; 7: 36-41Abstract Full Text Full Text PDF PubMed Scopus (353) Google Scholar Typically, miRNAs are capable of controlling the fate of cells in a time and tissue specific manner through regulation of cellular differentiation, developmental patterning, and morphogenesis. Therefore, manipulation of miRNAs could potentially be a useful approach to developing strategies for stem cell therapy. In this study, we investigated the role of miRNAs in an adherent subpopulation of CD34+ cells isolated from granulocyte colony-stimulating factor mobilized peripheral blood cells. We identified several clusters of miRNAs that were differentially expressed in these stem cells by miRNA profiling. Further analysis of one of the clusters revealed a miRNA that targets the stem cell gene Nanog. We show specifically that miR-181a* targets the 3′UTR of Nanog. Our data provide evidence that miR-181a* targets Nanog in a subpopulation of CD34+ cells. We performed miRNA profiling on a subpopulation of adherent CD34+ cells mobilized peripheral blood cells.16Gordon MY Levicar N Pai M Bachellier P Dimarakis I Al-Allaf F et al.Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor.Stem Cells. 2006; 24: 1822-1830Crossref PubMed Scopus (235) Google Scholar Using a 466 miRNA chip, we compared the miRNA signatures of adherent and nonadherent CD34+ cells (Figure 1a). These two subpopulations of CD34+ cells have previously been established as having distinct morphology, immunophenotype, and gene expression profile.16Gordon MY Levicar N Pai M Bachellier P Dimarakis I Al-Allaf F et al.Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor.Stem Cells. 2006; 24: 1822-1830Crossref PubMed Scopus (235) Google Scholar Moreover, the adherent CD34+ cells display greater plasticity than the nonadherent population as they have the potential to express determinants specific to liver, pancreas, heart, muscle, and nerve cell differentiation.16Gordon MY Levicar N Pai M Bachellier P Dimarakis I Al-Allaf F et al.Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor.Stem Cells. 2006; 24: 1822-1830Crossref PubMed Scopus (235) Google Scholar Hierarchical clustering revealed eight unique miRNA clusters that were differentially expressed in the adherent and nonadherent populations. This finding is not entirely surprising since the biological roles of miRNAs are known to vary according to their expression in distinct cell populations in normal tissues.17Lee SI Lee BR Hwang YS Lee HC Rengaraj D Song G et al.MicroRNA-mediated posttranscriptional regulation is required for maintaining undifferentiated properties of blastoderm and primordial germ cells in chickens.Proc Natl Acad Sci USA. 2011; 108: 10426-10431Crossref PubMed Scopus (61) Google Scholar,18Georgantas RW Hildreth R Morisot S Alder J Liu CG Heimfeld S et al.CD34+ hematopoietic stem-progenitor cell microRNA expression and function: a circuit diagram of differentiation control.Proc Natl Acad Sci USA. 2007; 104: 2750-2755Crossref PubMed Scopus (414) Google Scholar Interestingly, there were several miRNAs that have not been previously characterized for human in cluster 8 (Table 1). We were particularly interested in the miRNAs in cluster 8 because among the miRNAs in this cluster, miR-181a* (miR-181a-3p) was highly upregulated and studies have also shown that the miR-181 family are associated with regulation of CD34+ cells. However, the mRNA targets are largely unknown.18Georgantas RW Hildreth R Morisot S Alder J Liu CG Heimfeld S et al.CD34+ hematopoietic stem-progenitor cell microRNA expression and function: a circuit diagram of differentiation control.Proc Natl Acad Sci USA. 2007; 104: 2750-2755Crossref PubMed Scopus (414) Google Scholar,19Chen CZ Li L Lodish HF Bartel DP MicroRNAs modulate hematopoietic lineage differentiation.Science. 2004; 303: 83-86Crossref PubMed Scopus (2732) Google Scholar We did identify some conserved regions within the mature human miR-181 family of sequences which consist of miR-181a, miR-181b, miR-181c, and miR-181d (Figure 1b). We have taken the first step toward better understanding the role of miR-181a* in CD34+ cells.Table 1miRNAs from cluster 8 targets and functions in human stem cellsmiRNAPutative target(s)Known function(s)Refs.miR-191SOX4Helps the transition of epithelial cells into mesenchymal cells41He Y Cui Y Wang W Gu J Guo S Ma K et al.Hypomethylation of the hsa-miR-191 locus causes high expression of hsa-mir-191 and promotes the epithelial-to-mesenchymal transition in hepatocellular carcinoma.Neoplasia. 2011; 13: 841-853Abstract Full Text PDF PubMed Scopus (98) Google ScholarmiR-30cPAI-1, ALK2Enhances the differentiation of osteocytes and adipocytes; Controls mesenchymal and hematopoietic cell lineages progression42Karbiener M Neuhold C Opriessnig P Prokesch A Bogner-Strauss JG Scheideler M MicroRNA-30c promotes human adipocyte differentiation and co-represses PAI-1 and ALK2.RNA Biol. 2011; 8Crossref PubMed Scopus (105) Google Scholar, 43Merkerova M Belickova M Bruchova H Differential expression of microRNAs in hematopoietic cell lineages.Eur J Haematol. 2008; 81: 304-310Crossref PubMed Scopus (203) Google ScholarmiR-150NOTCH3Mediates T-cell development44Ghisi M Corradin A Basso K Frasson C Serafin V Mukherjee S et al.Modulation of microRNA expression in human T-cell development: targeting of NOTCH3 by miR-150.Blood. 2011; 117: 7053-7062Crossref PubMed Scopus (166) Google ScholarmiR-17-5pE2F1, BimRegulates stem cell differentiation and mediates embryonic development45Ho J Pandey P Schatton T Sims-Lucas S Khalid M Frank MH et al.The pro-apoptotic protein Bim is a microRNA target in kidney progenitors.J Am Soc Nephrol. 2011; 22: 1053-1063Crossref PubMed Scopus (74) Google Scholar, 46O’Donnell KA Wentzel EA Zeller KI Dang CV Mendell JT c-Myc-regulated microRNAs modulate E2F1 expression.Nature. 2005; 435: 839-843Crossref PubMed Scopus (2423) Google ScholarmiR-29aCDC24, TPM1, FZD5Plays an important role in stem cell differentiation; Regulates early heamatopoiesis47Bissels U Wild S Tomiuk S Hafner M Scheel H Mihailovic A et al.Combined characterization of microRNA and mRNA profiles delineates early differentiation pathways of CD133+ and CD34+ hematopoietic stem and progenitor cells.Stem Cells. 2011; 29: 847-857Crossref PubMed Scopus (73) Google Scholar, 48Han YC Park CY Bhagat G Zhang J Wang Y Fan JB et al.microRNA-29a induces aberrant self-renewal capacity in hematopoietic progenitors, biased myeloid development, and acute myeloid leukemia.J Exp Med. 2010; 207: 475-489Crossref PubMed Scopus (246) Google ScholarmiR-135bIBSP, OsterixModulates osteoblastic differentiation.49Schaap-Oziemlak AM Raymakers RA Bergevoet SM Gilissen C Jansen BJ Adema GJ et al.MicroRNA hsa-miR-135b regulates mineralization in osteogenic differentiation of human unrestricted somatic stem cells.Stem Cells Dev. 2010; 19: 877-885Crossref PubMed Scopus (79) Google ScholarmiR-196aHOXC8Plays a vital role in differentiation and proliferation of human adipose tissue-derived mesenchymal stem cells50Kim YJ Bae SW Yu SS Bae YC Jung JS miR-196a regulates proliferation and osteogenic differentiation in mesenchymal stem cells derived from human adipose tissue.J Bone Miner Res. 2009; 24: 816-825Crossref PubMed Scopus (200) Google ScholarmiR-199aAggrecan, SOX9, FABP4Regulates pluripotent stem cells (iPSCs) production; Mediates hMSC as well as chondrocyte differentiation51Wang J He Q Han C Gu H Jin L Li Q et al.p53-Facilitated miR-199a-3p Regulates Somatic Cell Reprogramming.Stem Cells. epub ahead of print, 2012Crossref Scopus (57) Google Scholar, 52Laine SK Alm JJ Virtanen SP Aro HT Laitala-Leinonen TK MicroRNAs miR-96, miR-124, and miR-199a regulate gene expression in human bone marrow-derived mesenchymal stem cells.J Cell Biochem. 2012; 113: 2687-2695Crossref PubMed Scopus (95) Google ScholarmiR-148bUnknownDifferentiation of hMSC53Schoolmeesters A Eklund T Leake D Vermeulen A Smith Q Force Aldred S et al.Functional profiling reveals critical role for miRNA in differentiation of human mesenchymal stem cells.PLoS ONE. 2009; 4: e5605Crossref PubMed Scopus (146) Google ScholarmiR-374UnknownPlays a role in neuronal progenitors transdifferentiated54Chang SJ Weng SL Hsieh JY Wang TY Chang MD Wang HW MicroRNA-34a modulates genes involved in cellular motility and oxidative phosphorylation in neural precursors derived from human umbilical cord mesenchymal stem cells.BMC Med Genomics. 2011; 4: 65Crossref PubMed Scopus (55) Google ScholarmiR-520cUnknownHepatocyte differentiation55Kim N Kim H Jung I Kim Y Kim D Han YM Expression profiles of miRNAs in human embryonic stem cells during hepatocyte differentiation.Hepatol Res. 2011; 41: 170-183Crossref PubMed Scopus (77) Google ScholarmiR-133aUnknownUnknownmiR-483UnknownUnknownmiR-411UnknownUnknownmiR-381UnknownUnknownmiR-515-5pUnknownUnknownmiR-107UnknownUnknownmiR-187UnknownUnknownmiR-296UnknownUnknownmiR-135aUnknownUnknownmiR-181a*UnknownUnknownAbbreviations: hMSC, human mesenchymal stem cells; miRNA, microRNA. Open table in a new tab Abbreviations: hMSC, human mesenchymal stem cells; miRNA, microRNA. We first determined whether miR-181a* affected the alkaline phosphatase activity in these cells. Higher alkaline phosphatase activity is generally associated with stem cells that are in a less differentiated state.20Ivanova N Dobrin R Lu R Kotenko I Levorse J DeCoste C et al.Dissecting self-renewal in stem cells with RNA interference.Nature. 2006; 442: 533-538Crossref PubMed Scopus (786) Google Scholar We found increasing levels of alkaline phosphatase activity in cells transfected with miR-181a* inhibitor when compared to transfection with miR-181a* mimic (Figure 1c). The data raises the possibility that these stem cell population could be maintained in a less differentiated state by inhibiting miR-181a* expression. We next applied an in-house genomic-bioinformatics database to identify and validate potential candidate target messages for miR-181a*. Being important factors in stem cell maintenance, the genomic loci of Nanog, STAT3, Hic1, Sox2, HoxB4, and Pou5f1 (Oct4) were scanned for putative target sites that was complementary to the miR-181a* sequence (5′-accaucgaccguugauuguacc-3′). Specifically, the annotated 3′ UTR sequences of these genes were scanned for seed sites with perfect reverse-complementarity to miRNA-181a* seed sequence (nucleotides 2–7 from the 5′ end) and sites with strong overall complementarity to miRNA-181a*. Of the putative target genes, we discovered that miR-181a* formed the most stable base pairing at the Nanog 3′UTR (Figure 2a). Since Nanog is an important regulator of stem cell maintenance and self-renewal,21Jaenisch R Young R Stem cells, the molecular circuitry of pluripotency and nuclear reprogramming.Cell. 2008; 132: 567-582Abstract Full Text Full Text PDF PubMed Scopus (1104) Google Scholar we determined whether miR-181a* targeted Nanog transcripts. Using endpoint reverse transcription-PCR (RT-PCR) (Figure 2b) and quantitative RT-PCR (Figure 2c) in transfected CD34+ stem cells, we found that inhibiting miR-181a* significantly upregulates transcript levels of Nanog. It has been reported that there exists a sensitive feedback loop that maintains the differentiation of stem cells through the control of essential stem cell genes such as Nanog. This loop enables where by environmental or molecular cues to influence the expression levels of genes, leading to a binary decision to switch downstream differentiation genes on or off.22Chickarmane V Troein C Nuber UA Sauro HM Peterson C Transcriptional dynamics of the embryonic stem cell switch.PLoS Comput Biol. 2006; 2: e123Crossref PubMed Scopus (212) Google Scholar The data raises the possibility that miR-181a* may be involved in directly regulating Nanog expression in CD34+ stem cells. To gain more insight into the functional interaction between miR-181a* and Nanog mRNA, we further interrogated the entire sequence alignment between the two RNA molecules. We discovered a high degree of base pairing at the 3′ region (7 nucleotides) and a weak base pairing at the 5′ seed site of miR-181a* (Figure 3a, top panel). In general most miRNA target sites have a strong base pairing at the 5′ seed site (7–8 nucleotides).11Bartel DP MicroRNAs: target recognition and regulatory functions.Cell. 2009; 136: 215-233Abstract Full Text Full Text PDF PubMed Scopus (15352) Google Scholar However, the 5′ seed site rule is not always the case and studies have shown that the 3′ region of miRNA also play an important role in targeting mRNA but are rare events.11Bartel DP MicroRNAs: target recognition and regulatory functions.Cell. 2009; 136: 215-233Abstract Full Text Full Text PDF PubMed Scopus (15352) Google Scholar These sites at the 3′ region of miRNAs are called 3′-supplementary and 3′-compensatory sites.11Bartel DP MicroRNAs: target recognition and regulatory functions.Cell. 2009; 136: 215-233Abstract Full Text Full Text PDF PubMed Scopus (15352) Google Scholar,23Brennecke J Stark A Russell RB Cohen SM Principles of microRNA-target recognition.PLoS Biol. 2005; 3: e85Crossref PubMed Scopus (1817) Google Scholar,24Brodersen P Voinnet O Revisiting the principles of microRNA target recognition and mode of action.Nat Rev Mol Cell Biol. 2009; 10: 141-148Crossref PubMed Scopus (534) Google Scholar In addition to the 3′ sites, a recent report has identified another site called the centre site25Shin C Nam JW Farh KK Chiang HR Shkumatava A Bartel DP Expanding the microRNA targeting code: functional sites with centered pairing.Mol Cell. 2010; 38: 789-802Abstract Full Text Full Text PDF PubMed Scopus (465) Google Scholar suggesting that miRNA targeting is more complex and diverse. In our case, because the base pairing at the 5′ seed site of miR-181a* was weak, we speculated that the strong complementarity pairing at the 3′ region would compensate and may follow the 3′-compensatory site rule similar to one of the earlier miRNA that was discovered such as let-711Bartel DP MicroRNAs: target recognition and regulatory functions.Cell. 2009; 136: 215-233Abstract Full Text Full Text PDF PubMed Scopus (15352) Google Scholar,26Reinhart BJ Slack FJ Basson M Pasquinelli AE Bettinger JC Rougvie AE et al.The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans.Nature. 2000; 403: 901-906Crossref PubMed Scopus (3654) Google Scholar (Figure 3a, bottom panel). Both miR-181a* and let-7 miRNAs, in this setting show a very similar overall structural identity in the way they bind to their targets. To determine whether the 3′ region of miR-181a* was important, we generated, using RNAhybrid modeling,26Reinhart BJ Slack FJ Basson M Pasquinelli AE Bettinger JC Rougvie AE et al.The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans.Nature. 2000; 403: 901-906Crossref PubMed Scopus (3654) Google Scholar,27Elbashir SM Lendeckel W Tuschl T RNA interference is mediated by 21- and 22-nucleotide RNAs.Genes Dev. 2001; 15: 188-200Crossref PubMed Scopus (2659) Google Scholar the most energy stable complementarity base pairing between miR-181a* and Nanog 3′ UTR mRNA (Figure 3b, top panel) as well as determining the best mutations that would disrupt the 3′ region base pairing. This was generated by replacing two bases at the 3′UTR of Nanog (Figure 3b, bottom panel). To test this modeling we generated a dual-luciferase reporter construct for miR-181a* and Nanog 3′ UTR mRNA. We inserted, upstream of the firefly luciferase gene, either the wild-type 3′UTR of Nanog or a mutant variant containing the two point mutations predicted to abolish miR-181a* targeting (Figure 3b, bottom panel). Transfecting the wild-type Nanog 3′UTR reporter construct by itself or with the miR-181a* mimic into CD34+ stem cells significantly reduced expression of luciferase (Figure 3c). Transfecting miR-181a* inhibitor caused a significant reversal in suppression of luciferase expression indicating that binding of miR-181a* to the 3′UTR of Nanog was suppressed thus allowing expression of the luciferase reporter. By contrast, transfection of the mutant Nanog 3′UTR reporter alone or in combination with miR-181a* mimic or inhibitor had no repressive effect on expression of the luciferase reporter (Figure 3c). This data therefore suggests that miR-181a* directly targets the 3′UTR region of Nanog. This is the first attempt to characterize miR-181a* in CD34+ cells and we have identified Nanog as its potential target. Understanding the cellular and molecular mechanisms responsible for the multi-pluripotency of stem cells will be critical for their practical use in therapy. MicroRNAs have been shown to be important and necessary for proper stem cell regulation.18Georgantas RW Hildreth R Morisot S Alder J Liu CG Heimfeld S et al.CD34+ hematopoietic stem-progenitor cell microRNA expression and function: a circuit diagram of differentiation control.Proc Natl Acad Sci USA. 2007; 104: 2750-2755Crossref PubMed Scopus (414) Google Scholar,28Hatfield SD Shcherbata HR Fischer KA Nakahara K Carthew RW Ruohola-Baker H Stem cell division is regulated by the microRNA pathway.Nature. 2005; 435: 974-978Crossref PubMed Scopus (576) Google Scholar,29Wulczyn FG Smirnova L Rybak A Brandt C Kwidzinski E Ninnemann O et al.Post-transcriptional regulation of the let-7 microRNA during neural cell specification.FASEB J. 2007; 21: 415-426Crossref PubMed Scopus (282) Google Scholar,30Wang Y Russell I Chen C MicroRNA and stem cell regulation.Curr Opin Mol Ther. 2009; 11: 292-298PubMed Google Scholar In this study, we generated a miRNA profile from a subpopulation of adherent CD34+ cells isolated from G- colony-stimulating factor mobilized peripheral blood and demonstrated a marked difference in expression patterns for a small cluster of miRNA. Using bioinformatics, we identified Nanog as a potential target for miR-181a*. Interestingly, there have been works published regarding other potential targets for the miR-181 family. For example, miR-181b has been shown to be an important regulator of nuclear factor-κB signaling in the vascular endothelium by targeting importin-α3.31Sun X Icli B Wara AK Belkin N He S Kobzik L et al.MicroRNA-181b regulates NF-?B-mediated vascular inflammation.J Clin Invest. 2012; 122: 1973-1990PubMed Google Scholar In another cell type, miR-181b has been shown to target Tcl1 oncogene in chronic lymphocytic leukemia.32Pekarsky Y Santanam U Cimmino A Palamarchuk A Efanov A Maximov V et al.Tcl1 expression in chronic lymphocytic leukemia is regulated by miR-29 and miR-181.Cancer Res. 2006; 66: 11590-11593Crossref PubMed Scopus (518) Google Scholar Similar to miR-181b, miR-181a also has several targets such as Prox1 in lymphatic endothelial cells, C2H2 zinc-finger proteins, and multiple phosphatases such as SHP2, PTPN22, DUSP5, and DUSP6.33Kazenwadel J Michael MZ Harvey NL Prox1 expression is negatively regulated by miR-181 in endothelial cells.Blood. 2010; 116: 2395-2401Crossref PubMed Scopus (122) Google Scholar,34Huang S Wu S Ding J Lin J Wei L Gu J et al.MicroRNA-181a modulates gene expression of zinc finger family members by directly targeting their coding regions.Nucleic Acids Res. 2010; 38: 7211-7218Crossref PubMed Scopus (67) Google Scholar,35Schnall-Levin M Rissland OS Johnston WK Perrimon N Bartel DP Berger B Unusually effective microRNA targeting within repeat-rich coding regions of mammalian mRNAs.Genome Res. 2011; 21: 1395-1403Crossref PubMed Scopus (99) Google Scholar,36Li QJ Chau J Ebert PJ Sylvester G Min H Liu G et al.miR-181a is an intrinsic modulator of T cell sensitivity and selection.Cell. 2007; 129: 147-161Abstract Full Text Full Text PDF PubMed Scopus (965) Google Scholar Although miR-181a*′s binding site within the 3′UTR of Nanog displayed a weak canonical 5′ miRNA seed pairing, it also showed strong complementarity at the miRNA 3′ end and at the central Ago2 catalytic site 10–11 nucleotides from the 5′ miRNA site.27Elbashir SM Lendeckel W Tuschl T RNA interference is mediated by 21- and 22-nucleotide RNAs.Genes Dev. 2001; 15: 188-200Crossref PubMed Scopus (2659) Google Scholar This binding pattern is similar with the “3′ compensatory site model previously identified with let7-miRNA.”11Bartel DP MicroRNAs: target recognition and regulatory functions.Cell. 2009; 136: 215-233Abstract Full Text Full Text PDF PubMed Scopus (15352) Google Scholar,23Brennecke J Stark A Russell RB Cohen SM Principles of microRNA-target recognition.PLoS Biol. 2005; 3: e85Crossref PubMed Scopus (1817) Google Scholar Importantly, by introducing two point mutations to disrupt the 3′ and central complementary sites, we abolished miR-181a* regulation of a reporter construct containing the 3′UTR of Nanog. Our studies highlight a new stem cell-related target for the miR-181 family and show that miR-181a* directly targets Nanog. Isolation and growth of a CD34+ stem cell population. The hematopoietic blood samples were obtained with informed patient consent and approved by the Hammersmith Hospital Research Ethics Committee. Samples of granulocyte-colony stimulating factor mobilized peripheral blood progenitor cells were processed by leukapheresis at the Stem Cell Laboratory at the Hammersmith Hospital. Briefly, human mobilized peripheral blood samples were diluted in a ratio of 1:4 in Hanks’-buffered saline solution (Gibco, Paisley, UK), the mononuclear cells were separated by centrifugation over a Lymphoprep (Dundee, Scotland) density gradient at 1,800 rpm for 30 minutes. The mononuclear cell fraction at the interface was aspirated and washed twice with Hanks’-buffered saline solution, and finally with MACS buffer (phosphate-buffered saline solution) at pH 7.2 supplemented with 0.5% bovine serum albumin and 2 mmol/l EDTA). CD34+ cells were isolated using a CD34+ isolation kit (Miltenyi Biotec, Cologne, Germany) according to the manufacturer's protocol. Adherent CD34+ progenitor cells were isolated as previously described with modification.16Gordon MY Levicar N Pai M Bachellier P Dimarakis I Al-Allaf F et al.Characterization and clinical application of human CD34+ stem/progenitor cell populations mobilized into the blood by granulocyte colony-stimulating factor.Stem Cells. 2006; 24: 1822-1830Crossref PubMed Scopus (235) Google Scholar Briefly, isolated CD34+ cells were added to 24-well or 35-mm tissue-culture treated dishes (Nunc, Roskilde, Denmark) at a density of 2.5–5 × 105 cells in α-minimum essential medium to isolate the adherent CD34+ cell population. After 30-minute" @default.
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• W2061457029 title "MicroRNA-181a* Targets Nanog in a Subpopulation of CD34+ Cells Isolated From Peripheral Blood" @default.
• W2061457029 cites W126539919 @default.
• W2061457029 cites W1616307661 @default.
• W2061457029 cites W1920303762 @default.
• W2061457029 cites W1952134382 @default.
• W2061457029 cites W1970291858 @default.
• W2061457029 cites W1974560023 @default.
• W2061457029 cites W1975200987 @default.
• W2061457029 cites W1993174948 @default.
• W2061457029 cites W1993640536 @default.
• W2061457029 cites W1993799748 @default.
• W2061457029 cites W1995073275 @default.
• W2061457029 cites W1997189773 @default.
• W2061457029 cites W2004836344 @default.
• W2061457029 cites W2006169630 @default.
• W2061457029 cites W2013646486 @default.
• W2061457029 cites W2025567420 @default.
• W2061457029 cites W2030621748 @default.
• W2061457029 cites W2034412957 @default.
• W2061457029 cites W2034885353 @default.
• W2061457029 cites W2036795046 @default.
• W2061457029 cites W2037205041 @default.
• W2061457029 cites W2054721944 @default.
• W2061457029 cites W2054923580 @default.
• W2061457029 cites W2059382687 @default.
• W2061457029 cites W2060245619 @default.
• W2061457029 cites W2067227192 @default.
• W2061457029 cites W2069337669 @default.
• W2061457029 cites W2069923423 @default.
• W2061457029 cites W2073745960 @default.
• W2061457029 cites W2075291842 @default.
• W2061457029 cites W2077066570 @default.
• W2061457029 cites W2084848143 @default.
• W2061457029 cites W2088930981 @default.
• W2061457029 cites W2094253263 @default.
• W2061457029 cites W2095252059 @default.
• W2061457029 cites W2095509700 @default.
• W2061457029 cites W2103054294 @default.
• W2061457029 cites W2103546044 @default.
• W2061457029 cites W2108241558 @default.
• W2061457029 cites W2108871359 @default.
• W2061457029 cites W2110043783 @default.
• W2061457029 cites W2117391818 @default.
• W2061457029 cites W2123137429 @default.
• W2061457029 cites W2134427620 @default.
• W2061457029 cites W2149784145 @default.
• W2061457029 cites W2152993806 @default.
• W2061457029 cites W2154699417 @default.
• W2061457029 cites W2155649150 @default.
• W2061457029 cites W2159999899 @default.
• W2061457029 cites W2160387598 @default.
• W2061457029 cites W2162674813 @default.
• W2061457029 cites W2163128475 @default.
• W2061457029 cites W2171488801 @default.
• W2061457029 cites W256843741 @default.
• W2061457029 cites W2910065277 @default.
• W2061457029 cites W4256341283 @default.
• W2061457029 doi "https://doi.org/10.1038/mtna.2012.29" @default.
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