title: Hsa-miR-520d converts fibroblasts into CD105+ populations.
abstract: BACKGROUND
  We have previously shown that hsa-miR-520d-5p can convert cancer cells into induced pluripotent stem cells (iPSCs) or mesenchymal stem cells (MSCs) via a dedifferentiation by a demethylation mechanism.
  
  
  METHODS
  We tested the effect of miR-520d-5p on human fibroblasts to determine whether it could be safely used in normal cells for future clinical therapeutic applications. After we transfected the microRNA into fibroblasts, we analyzed the phenotypic changes, gene expression levels, and stemness induction in vitro, and we evaluated tumor formation in an in vivo xenograft model.
  
  
  RESULTS
  The transfected fibroblasts turned into CD105+ cell populations, survived approximately 24 weeks, and exhibited increases in both the collagen-producing ability and in differentiation. Combinatorial transfection of small interfering RNAs for miR-520d-5p target genes (ELAVL2, GATAD2B, and TEAD1) produced similar results to miR-520d-5p transfection. These molecules converted normal cells into MSCs and not iPSCs.
  
  
  CONCLUSIONS
  In vitro data indicate the potent usefulness of this small molecule as a therapeutic biomaterial in normal cells and cancer cells because CD105+ cells never converted to iPSCs despite repeated transfections and all types of transfectants lost their tumorigenicity. This maintenance of a benign state following miR-520d-5p transfection appears to be caused by p53 upregulation. We conclude that miR-520d-5p may be a useful biomaterial at an in vitro level.
authors.affiliation: Division of Pharmacotherapeutics, Department of Pathophysiological and Therapeutic Science, Faculty of Medicine, Tottori University, Tottori, 683-8503, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Ishihara
authors.forename: Satoshi
authors.initials: S
authors.lastname: Tsuno
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kuwamoto
authors.forename: Taro
authors.initials: T
authors.lastname: Yamashita
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Endo
authors.forename: Junichi
authors.initials: J
authors.lastname: Hasegawa
authors.forename: Norimasa
authors.initials: N
authors.lastname: Miura
mesh_terms: D015703:Antigens, CD / Q000276:immunology*; D015197:Carcinogenicity Tests; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival; D003094:Collagen / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors; D000067777:ELAV-Like Protein 2 / Q000037:antagonists & inhibitors; D000071063:Endoglin; D005347:Fibroblasts / Q000187:drug effects* / Q000276:immunology; D050980:GATA Transcription Factors / Q000037:antagonists & inhibitors; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects; D059630:Mesenchymal Stem Cells / Q000187:drug effects; D035683:MicroRNAs / Q000494:pharmacology*; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors; D010641:Phenotype; D034741:RNA, Small Interfering / Q000494:pharmacology; D011956:Receptors, Cell Surface / Q000276:immunology*; D012097:Repressor Proteins; D000091262:TEA Domain Transcription Factors; D059505:Telomere Homeostasis / Q000187:drug effects; D014157:Transcription Factors / Q000037:antagonists & inhibitors
chemical_list: D015703:Antigens, CD; D004268:DNA-Binding Proteins; D000067777:ELAV-Like Protein 2; C086811:ELAVL2 protein, human; C064206:ENG protein, human; D000071063:Endoglin; D050980:GATA Transcription Factors; C468770:GATAD2B protein, human; C575048:MIRN520 microRNA, human; D035683:MicroRNAs; D009687:Nuclear Proteins; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface; D012097:Repressor Proteins; D000091262:TEA Domain Transcription Factors; C057182:TEAD1 protein, human; D014157:Transcription Factors; D003094:Collagen