Ablation of the miRNA Cluster 24 Has Profound Effects on Extracellular Matrix Protein Abundance in Cartilage

Int J Mol Sci. 2020 Jun 9;21(11):4112. doi: 10.3390/ijms21114112.

Abstract

MicroRNAs (miRNAs) regulate cartilage differentiation and contribute to the onset and progression of joint degeneration. These small RNA molecules may affect extracellular matrix organization (ECM) in cartilage, but for only a few miRNAs has this role been defined in vivo. Previously, we showed that cartilage-specific genetic ablation of the Mirc24 cluster in mice leads to impaired cartilage development due to increased RAF/MEK/ERK pathway activation. Here, we studied the expression of the cluster in cartilage by LacZ reporter gene assays and determined its role for extracellular matrix homeostasis by proteome and immunoblot analysis. The cluster is expressed in prehypertrophic/hypertrophic chondrocytes of the growth plate and we now show that the cluster is also highly expressed in articular cartilage. Cartilage-specific loss of the cluster leads to increased proteoglycan 4 and matrix metallopeptidase 13 levels and decreased aggrecan and collagen X levels in epiphyseal cartilage. Interestingly, these changes are linked to a decrease in SRY-related HMG box-containing (SOX) transcription factors 6 and 9, which regulate ECM production in chondrocytes. Our data suggests that the Mirc24 cluster is important for ECM homoeostasis and the expression of transcriptional regulators of matrix production in cartilage.

Keywords: MMP13; Mirc24; PRG4; SOX6; SOX9; articular; cartilage; extracellular matrix; miR-322; miR-503.

MeSH terms

  • Animals
  • Cartilage, Articular / metabolism*
  • Cartilage, Articular / physiology
  • Collagen Type II / genetics
  • Extracellular Matrix Proteins / genetics*
  • Extracellular Matrix Proteins / metabolism
  • Gene Expression Regulation
  • Growth Plate / chemistry
  • Male
  • Matrix Metalloproteinase 13 / genetics
  • Matrix Metalloproteinase 13 / metabolism
  • Mice, Transgenic
  • MicroRNAs / genetics*
  • Multigene Family
  • Osteoarthritis / genetics*
  • Proteoglycans / genetics
  • Proteoglycans / metabolism
  • SOX9 Transcription Factor / genetics
  • SOX9 Transcription Factor / metabolism
  • SOXD Transcription Factors / genetics
  • SOXD Transcription Factors / metabolism

Substances

  • Col2a1 protein, mouse
  • Collagen Type II
  • Extracellular Matrix Proteins
  • MicroRNAs
  • Prg4 protein, mouse
  • Proteoglycans
  • SOX9 Transcription Factor
  • SOXD Transcription Factors
  • Sox6 protein, mouse
  • Sox9 protein, mouse
  • Matrix Metalloproteinase 13
  • Mmp13 protein, mouse