title: Genome-Wide MicroRNA and Gene Analysis of Mesenchymal Stem Cell Chondrogenesis Identifies an Essential Role and Multiple Targets for miR-140-5p.
abstract: microRNAs (miRNAs) are abundantly expressed in development where they are critical determinants of cell differentiation and phenotype. Accordingly miRNAs are essential for normal skeletal development and chondrogenesis in particular. However, the question of which miRNAs are specific to the chondrocyte phenotype has not been fully addressed. Using microarray analysis of miRNA expression during mesenchymal stem cell chondrogenic differentiation and detailed examination of the role of essential differentiation factors, such as SOX9, TGF-β, and the cell condensation phase, we characterize the repertoire of specific miRNAs involved in chondrocyte development, highlighting in particular miR-140 and miR-455. Further with the use of mRNA microarray data we integrate miRNA expression and mRNA expression during chondrogenesis to underline the particular importance of miR-140, especially the -5p strand. We provide a detailed identification and validation of direct targets of miR-140-5p in both chondrogenesis and adult chondrocytes with the use of microarray and 3'UTR analysis. This emphasizes the diverse array of targets and pathways regulated by miR-140-5p. We are also able to confirm previous experimentally identified targets but, additionally, identify a novel positive regulation of the Wnt signaling pathway by miR-140-5p. Wnt signaling has a complex role in chondrogenesis and skeletal development and these findings illustrate a previously unidentified role for miR-140-5p in regulation of Wnt signaling in these processes. Together these developments further highlight the role of miRNAs during chondrogenesis to improve our understanding of chondrocyte development and guide cartilage tissue engineering.
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Matt J
authors.initials: MJ
authors.lastname: Barter
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Maria
authors.initials: M
authors.lastname: Tselepi
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Gómez
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Steven
authors.initials: S
authors.lastname: Woods
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Wang
authors.initials: W
authors.lastname: Hui
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Graham R
authors.initials: GR
authors.lastname: Smith
authors.affiliation: Institute for Ageing and Health, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Daryl P
authors.initials: DP
authors.lastname: Shanley
authors.affiliation: School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.
authors.forename: Ian M
authors.initials: IM
authors.lastname: Clark
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: David A
authors.initials: DA
authors.lastname: Young
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000502:physiology*; D020869:Gene Expression Profiling / Q000379:methods*; D018390:Gene Targeting / Q000379:methods*; D055106:Genome-Wide Association Study / Q000379:methods*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D035683:MicroRNAs / Q000502:physiology*; D008875:Middle Aged; D055815:Young Adult
chemical_list: D035683:MicroRNAs; C544302:Mirn140 microRNA, human
keywords: Chondrogenesis; Differentiation; Epigenetics; Gene expression; Mesenchymal stem cells; miRNA