title: MicroRNA-140 is expressed in differentiated human articular chondrocytes and modulates interleukin-1 responses.
abstract: OBJECTIVE
  MicroRNA (miRNA) are a class of noncoding small RNAs that act as negative regulators of gene expression. MiRNA exhibit tissue-specific expression patterns, and changes in their expression may contribute to pathogenesis. The objectives of this study were to identify miRNA expressed in articular chondrocytes, to determine changes in osteoarthritic (OA) cartilage, and to address the function of miRNA-140 (miR-140).
  
  
  METHODS
  To identify miRNA specifically expressed in chondrocytes, we performed gene expression profiling using miRNA microarrays and quantitative polymerase chain reaction with human articular chondrocytes compared with human mesenchymal stem cells (MSCs). The expression pattern of miR-140 was monitored during chondrogenic differentiation of human MSCs in pellet cultures and in human articular cartilage from normal and OA knee joints. We tested the effects of interleukin-1beta (IL-1beta) on miR-140 expression. Double-stranded miR-140 (ds-miR-140) was transfected into chondrocytes to analyze changes in the expression of genes associated with OA.
  
  
  RESULTS
  Microarray analysis showed that miR-140 had the largest difference in expression between chondrocytes and MSCs. During chondrogenesis, miR-140 expression in MSC cultures increased in parallel with the expression of SOX9 and COL2A1. Normal human articular cartilage expressed miR-140, and this expression was significantly reduced in OA tissue. In vitro treatment of chondrocytes with IL-1beta suppressed miR-140 expression. Transfection of chondrocytes with ds-miR-140 down-regulated IL-1beta-induced ADAMTS5 expression and rescued the IL-1beta-dependent repression of AGGRECAN gene expression.
  
  
  CONCLUSIONS
  This study shows that miR-140 has a chondrocyte differentiation-related expression pattern. The reduction in miR-140 expression in OA cartilage and in response to IL-1beta may contribute to the abnormal gene expression pattern characteristic of OA.
authors.affiliation: Department of Molecular and Experimental Medicine, Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Miyaki
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Nakasa
authors.forename: Shuhei
authors.initials: S
authors.lastname: Otsuki
authors.forename: Shawn P
authors.initials: SP
authors.lastname: Grogan
authors.forename: Reiji
authors.initials: R
authors.lastname: Higashiyama
authors.forename: Atsushi
authors.initials: A
authors.lastname: Inoue
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Kato
authors.forename: Tempei
authors.initials: T
authors.lastname: Sato
authors.forename: Martin K
authors.initials: MK
authors.lastname: Lotz
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Asahara
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism; D000071116:ADAMTS5 Protein; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D053674:Aggrecans / Q000378:metabolism; D002358:Cartilage, Articular / Q000378:metabolism* / Q000473:pathology; D016022:Case-Control Studies; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D024043:Collagen Type II / Q000378:metabolism; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism* / Q000494:pharmacology; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000473:pathology; D035683:MicroRNAs / Q000378:metabolism*; D020370:Osteoarthritis, Knee / Q000378:metabolism* / Q000473:pathology; D012330:RNA, Double-Stranded / Q000378:metabolism*; D055758:SOX9 Transcription Factor / Q000378:metabolism; D014162:Transfection
chemical_list: D053674:Aggrecans; C499177:COL2A1 protein, human; D024043:Collagen Type II; D053583:Interleukin-1beta; D035683:MicroRNAs; C544302:Mirn140 microRNA, human; D012330:RNA, Double-Stranded; D055758:SOX9 Transcription Factor; C527932:SOX9 protein, human; D051722:ADAM Proteins; D000071116:ADAMTS5 Protein; C121532:ADAMTS5 protein, human