Genome-Wide MicroRNA and Gene Analysis of Mesenchymal Stem Cell Chondrogenesis Identifies an Essential Role and Multiple Targets for miR-140-5p.
Genome-Wide MicroRNA and Gene Analysis of Mesenchymal Stem Cell Chondrogenesis Identifies an Essential Role and Multiple Targets for miR-140-5p.
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DOI：
10.1002/stem.2093
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发表时间：
2015-11
期刊：
影响因子：
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通讯作者：
Young DA
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文献类型：
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作者：
Barter MJ;Tselepi M;Gómez R;Woods S;Hui W;Smith GR;Shanley DP;Clark IM;Young DA
microRNAs （miRNAs）在发育过程中大量表达，是细胞分化和表型的关键决定因素。因此，mirna对正常骨骼发育和软骨形成至关重要。然而，哪些mirna对软骨细胞表型特异性的问题尚未得到充分解决。通过对间充质干细胞成软骨分化过程中miRNA表达的微阵列分析，以及对必需分化因子（如SOX9、TGF - β和细胞凝聚期）作用的详细检查，我们描述了参与软骨细胞发育的特定miRNA，特别是miR‐140和miR‐455。此外，通过使用mRNA微阵列数据，我们整合了软骨形成过程中miRNA表达和mRNA表达，以强调miR - 140，特别是- 5p链的特殊重要性。我们使用微阵列和3'UTR分析详细鉴定和验证了miR - 140 - 5p在软骨形成和成体软骨细胞中的直接靶点。这强调了miR - 140 - 5p调控的靶点和通路的多样性。我们也能够确认先前实验确定的靶标，此外，我们还确定了miR‐140‐5p对Wnt信号通路的一种新的正调控。Wnt信号在软骨形成和骨骼发育中具有复杂的作用，这些发现说明了miR - 140 - 5p在这些过程中调控Wnt信号的作用。总之，这些进展进一步强调了mirna在软骨形成过程中的作用，以提高我们对软骨细胞发育的理解并指导软骨组织工程。干细胞2015；33:3266 - 3280
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. Stem Cells 2015;33:3266–3280