title: MicroRNA expression profiles of human iPSCs differentiation into insulin-producing cells.
abstract: OBJECTIVE
  MicroRNAs are a class of small noncoding RNAs, which control gene expression by inhibition of mRNA translation. MicroRNAs are involved in the control of biological processes including cell differentiation. Here, we aim at characterizing microRNA expression profiles during differentiation of human induced pluripotent stem cells (hiPSCs) into insulin-producing cells.
  
  
  METHODS
  We differentiated hiPSCs toward endocrine pancreatic lineage following a 18-day protocol. We analyzed genes and microRNA expression levels using RT real-time PCR and TaqMan microRNA arrays followed by bioinformatic functional analysis.
  
  
  RESULTS
  MicroRNA expression profiles analysis of undifferentiated hiPSCs during pancreatic differentiation revealed that 347/768 microRNAs were expressed at least in one time point of all samples. We observed 18 microRNAs differentially expressed: 11 were upregulated (miR-9-5p, miR-9-3p, miR-10a, miR-99a-3p, miR-124a, miR-135a, miR-138, miR-149, miR-211, miR-342-3p and miR-375) and 7 downregulated (miR-31, miR-127, miR-143, miR-302c-3p, miR-373, miR-518b and miR-520c-3p) during differentiation into insulin-producing cells. Selected microRNAs were further evaluated during differentiation of Sendai-virus-reprogrammed hiPSCs using an improved endocrine pancreatic beta cell derivation protocol and, moreover, in differentiated NKX6.1+sorted cells. Following Targetscan7.0 analysis of target genes of differentially expressed microRNAs and gene ontology classification, we found that such target genes belong to categories of major significance in pancreas organogenesis and development or exocytosis.
  
  
  CONCLUSIONS
  We detected a specific hiPSCs microRNAs signature during differentiation into insulin-producing cells and demonstrated that differentially expressed microRNAs target several genes involved in pancreas organogenesis.
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy.
authors.forename: Guido
authors.initials: G
authors.lastname: Sebastiani
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Valentini
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy.
authors.forename: Giuseppina Emanuela
authors.initials: GE
authors.lastname: Grieco
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy.
authors.forename: Giuliana
authors.initials: G
authors.lastname: Ventriglia
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Nigi
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Mancarella
authors.affiliation: Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Pellegrini
authors.affiliation: Division of Neuroscience, Institute of Experimental Neurology (INSpe), IRCCS San Raffaele Hospital, Milan, Italy.
authors.forename: Gianvito
authors.initials: G
authors.lastname: Martino
authors.affiliation: Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Sordi
authors.affiliation: Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Piemonti
authors.affiliation: Diabetes Unit, Department of Medicine, Surgery and Neuroscience, University of Siena, Siena, Italy. francesco.dotta@alice.it.|Fondazione Umberto di Mario ONLUS, Toscana Life Sciences, Siena, Italy. francesco.dotta@alice.it.
authors.forename: Francesco
authors.initials: F
authors.lastname: Dotta
mesh_terms: D000328:Adult; D000368:Aged; D002454:Cell Differentiation / Q000235:genetics*; D002478:Cells, Cultured; D020869:Gene Expression Profiling; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D007328:Insulin / Q000378:metabolism; D050417:Insulin-Secreting Cells / Q000378:metabolism / Q000502:physiology*; D007515:Islets of Langerhans / Q000378:metabolism / Q000502:physiology; D035683:MicroRNAs / Q000235:genetics*; D046228:Microarray Analysis; D008875:Middle Aged; D010179:Pancreas / Q000378:metabolism; D059467:Transcriptome
chemical_list: D007328:Insulin; D035683:MicroRNAs
keywords: Diabetes; Insulin-producing cells; iPSC; microRNAs