MicroRNA-140 is expressed in differentiated human articular chondrocytes and modulates interleukin-1 responses.

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. CONCLUSION 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.

[1]  C. Burge,et al.  Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets , 2005, Cell.

[2]  V. Ambros,et al.  An Extensive Class of Small RNAs in Caenorhabditis elegans , 2001, Science.

[3]  D. D’Lima,et al.  Cell death in cartilage. , 2004, Osteoarthritis and cartilage.

[4]  V. Lefebvre,et al.  Modulation by interleukin 1 and tumor necrosis factor alpha of production of collagenase, tissue inhibitor of metalloproteinases and collagen types in differentiated and dedifferentiated articular chondrocytes. , 1990, Biochimica et biophysica acta.

[5]  Molecular network of cartilage homeostasis and osteoarthritis , 2008, Medicinal research reviews.

[6]  M. Lotz.  Cytokines in cartilage injury and repair. , 2001, Clinical orthopaedics and related research.

[7]  H. Horvitz,et al.  MicroRNA Expression in Zebrafish Embryonic Development , 2005, Science.

[8]  N. Rajewsky,et al.  A pancreatic islet-specific microRNA regulates insulin secretion , 2004, Nature.

[9]  T. Dalmay,et al.  The cartilage specific microRNA‐140 targets histone deacetylase 4 in mouse cells , 2006, FEBS letters.

[10]  S. Krane,et al.  Interleukin 1 suppresses expression of cartilage-specific types II and IX collagens and increases types I and III collagens in human chondrocytes. , 1988, The Journal of clinical investigation.

[11]  T. Aigner,et al.  Mechanisms of Disease: role of chondrocytes in the pathogenesis of osteoarthritis—structure, chaos and senescence , 2007, Nature Clinical Practice Rheumatology.

[12]  M. Goldring.  Update on the biology of the chondrocyte and new approaches to treating cartilage diseases. , 2006, Best practice & research. Clinical rheumatology.

[13]  E. Olson,et al.  A signature pattern of stress-responsive microRNAs that can evoke cardiac hypertrophy and heart failure , 2006, Proceedings of the National Academy of Sciences.

[14]  S. Miyaki,et al.  Expression of MicroRNA-146a in osteoarthritis cartilage. , 2009, Arthritis and rheumatism.

[15]  J. Dudhia.  Aggrecan, aging and assembly in articular cartilage , 2005, Cellular and Molecular Life Sciences CMLS.

[16]  L. Lim,et al.  An Abundant Class of Tiny RNAs with Probable Regulatory Roles in Caenorhabditis elegans , 2001, Science.

[17]  T. Tuschl,et al.  Identification of Novel Genes Coding for Small Expressed RNAs , 2001, Science.

[18]  D. Bartel.  MicroRNAs Genomics, Biogenesis, Mechanism, and Function , 2004, Cell.

[19]  R. Maier,et al.  Interleukin-11, an inducible cytokine in human articular chondrocytes and synoviocytes, stimulates the production of the tissue inhibitor of metalloproteinases. , 1993, The Journal of biological chemistry.

[20]  G. Nuovo,et al.  Experimental validation of miRNA targets. , 2008, Methods.

[21]  Keiichiro Nishida,et al.  Expression of microRNA-146 in rheumatoid arthritis synovial tissue. , 2008, Arthritis and rheumatism.

[22]  H. Dorfman,et al.  Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. , 1971, The Journal of bone and joint surgery. American volume.

[23]  Michael Detmar,et al.  Altered expression of MicroRNA in synovial fibroblasts and synovial tissue in rheumatoid arthritis. , 2008, Arthritis and rheumatism.

[24]  A. Fourie,et al.  ADAMTS5 is the major aggrecanase in mouse cartilage in vivo and in vitro , 2005, Nature.

[25]  K. Geoghegan,et al.  Cloning, expression, and type II collagenolytic activity of matrix metalloproteinase-13 from human osteoarthritic cartilage. , 1996, The Journal of clinical investigation.

[26]  M. Goldring,et al.  Osteoarthritis and cartilage: The role of cytokines , 2000, Current rheumatology reports.

[27]  M. Goldring,et al.  The role of the chondrocyte in osteoarthritis. , 2000, Arthritis and rheumatism.

[28]  A. McMahon,et al.  Dicer-dependent pathways regulate chondrocyte proliferation and differentiation , 2008, Proceedings of the National Academy of Sciences.

[29]  L. Joosten,et al.  TNF-induced structural joint damage is mediated by IL-1 , 2007, Proceedings of the National Academy of Sciences.

[30]  M. Tortorella,et al.  Inhibition of ADAM-TS4 and ADAM-TS5 Prevents Aggrecan Degradation in Osteoarthritic Cartilage* , 2002, The Journal of Biological Chemistry.

[31]  Atlanta,et al.  Estimates of the prevalence of arthritis and other rheumatic conditions in the United States. Part I. , 2008, Arthritis and rheumatism.

[32]  Michael T. McManus,et al.  The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[33]  F. Slack,et al.  Oncomirs — microRNAs with a role in cancer , 2006, Nature Reviews Cancer.

[34]  S. Gabriel,et al.  Estimates of the prevalence of arthritis and other rheumatic conditions in the United States. Part II. , 2008, Arthritis and rheumatism.

[35]  J. Postlethwait,et al.  MicroRNA Mirn140 modulates Pdgf signaling during palatogenesis , 2008, Nature Genetics.

[36]  M. Lotz,et al.  Repression of chondrogenesis through binding of notch signaling proteins HES-1 and HEY-1 to N-box domains in the COL2A1 enhancer site. , 2008, Arthritis and rheumatism.

[37]  Konstantinos N. Malizos,et al.  Integrative MicroRNA and Proteomic Approaches Identify Novel Osteoarthritis Genes and Their Collaborative Metabolic and Inflammatory Networks , 2008, PloS one.

[38]  T. Aigner,et al.  Relative messenger RNA expression profiling of collagenases and aggrecanases in human articular chondrocytes in vivo and in vitro. , 2002, Arthritis and rheumatism.

[39]  C. Little,et al.  Blocking aggrecanase cleavage in the aggrecan interglobular domain abrogates cartilage erosion and promotes cartilage repair. , 2007, The Journal of clinical investigation.