title: miR-10a restores human mesenchymal stem cell differentiation by repressing KLF4.
abstract: miRNAs have recently been shown to play a significant role in human aging. However, data demonstrating the effects of aging-related miRNAs in human mesenchymal stem cells (hMSCs) are limited. We observed that hMSC differentiation decreased with aging. We also identified that miR-10a expression was significantly decreased with age by comparing the miRNA expression of hMSCs derived from young and aged individuals. Therefore, we hypothesized that the downregulation of miR-10a may be associated with the decreased differentiation capability of hMSCs from aged individuals. Lentiviral constructs were used to up- or downregulate miR-10a in young and old hMSCs. Upregulation of miR-10a resulted in increased differentiation to adipogenic, osteogenic, and chondrogenic lineages and in reduced cell senescence. Conversely, downregulation of miR-10a resulted in decreased cell differentiation and increased cell senescence. A chimeric luciferase reporter system was generated, tagged with the full-length 3'-UTR region of KLF4 harboring the seed-matched sequence with or without four nucleotide mutations. These constructs were cotransfected with the miR-10a mimic into cells. The luciferase activity was significantly repressed by the miR-10a mimic, proving the direct binding of miR-10a to the 3'-UTR of KLF4. Direct suppression of KLF4 in aged hMSCs increased cell differentiation and decreased cell senescence. In conclusion, miR-10a restores the differentiation capability of aged hMSCs through repression of KLF4. Aging-related miRNAs may have broad applications in the restoration of cell dysfunction caused by aging.
authors.affiliation: Department of Cardiology, the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou Institute of Cardiovascular Disease, Guangzhou, China.
authors.forename: Jiao
authors.initials: J
authors.lastname: Li
authors.forename: Jun
authors.initials: J
authors.lastname: Dong
authors.forename: Zhen-Hui
authors.initials: ZH
authors.lastname: Zhang
authors.forename: Dong-Cheng
authors.initials: DC
authors.lastname: Zhang
authors.forename: Xiang-Yu
authors.initials: XY
authors.lastname: You
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhong
authors.forename: Min-Sheng
authors.initials: MS
authors.lastname: Chen
authors.forename: Shi-Ming
authors.initials: SM
authors.lastname: Liu
mesh_terms: D020413:3' Untranslated Regions; D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002454:Cell Differentiation / Q000502:physiology; D016922:Cellular Senescence / Q000235:genetics; D015536:Down-Regulation; D005260:Female; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics* / Q000378:metabolism; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism*; D009154:Mutation; D009711:Nucleotides / Q000235:genetics; D015854:Up-Regulation; D055815:Young Adult
chemical_list: D020413:3' Untranslated Regions; C000715467:KLF4 protein, human; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C547183:MIRN10 microRNA, human; D035683:MicroRNAs; D009711:Nucleotides