Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein

Arthritis Res Ther. 2013 Oct 4;15(5):R147. doi: 10.1186/ar4330.

Abstract

Introduction: This study aimed to characterize the glycophenotype of osteoarthritic cartilage and human chondrocytes.

Methods: Articular knee cartilage was obtained from nine osteoarthritis (OA) patients. mRNA levels for 27 glycosyltransferases were analyzed in OA chondrocytes using RT-qPCR. Additionally, N- and O-glycans were quantified using mass-spectrometry. Histologically, two cartilage areas with Mankin scores (MS) either ≤ 4 or ≥ 9 were selected from each patient representing areas of mild and severe OA, respectively. Tissue sections were stained with (1) a selected panel of plant lectins for probing into the OA glycophenotype, (2) the human lectins galectins-1 and -3, and (3) the glycoprotein asialofetuin (ASF) for visualizing β-galactoside-specific endogenous lectins.

Results: We found that OA chondrocytes expressed oligomannosidic structures as well as non-, mono- and disialylated complex-type N-glycans, and core 2 O-glycans. Reflecting B4GALNT3 mRNA presence in OA chondrocytes, LacdiNAc-terminated structures were detected. Staining profiles for plant and human lectins were dependent on the grade of cartilage degeneration, and ASF-positive cells were observed in significantly higher rates in areas of severe degeneration.

Conclusions: In summary, distinct aspects of the glycome in OA cartilage are altered with progressing degeneration. In particular, the alterations measured by galectin-3 and the pan-galectin sensor ASF encourage detailed studies of galectin functionality in OA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Asialoglycoproteins / chemistry
  • Asialoglycoproteins / metabolism
  • Cartilage, Articular / metabolism*
  • Cartilage, Articular / pathology
  • Cells, Cultured
  • Chondrocytes / metabolism*
  • Chondrocytes / pathology
  • Fetuins / chemistry
  • Fetuins / metabolism
  • Galectin 1 / metabolism
  • Galectin 3 / metabolism
  • Gene Expression
  • Glycoproteins / chemistry
  • Glycoproteins / genetics
  • Glycoproteins / metabolism*
  • Glycosyltransferases / genetics
  • Glycosyltransferases / metabolism
  • Histocytochemistry
  • Humans
  • Lectins / metabolism*
  • Mass Spectrometry
  • Molecular Structure
  • N-Acetylgalactosaminyltransferases / genetics
  • N-Acetylgalactosaminyltransferases / metabolism
  • Osteoarthritis / genetics
  • Osteoarthritis / metabolism*
  • Plant Lectins / metabolism
  • Polysaccharides / chemistry
  • Polysaccharides / metabolism
  • Protein Binding
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Asialoglycoproteins
  • Fetuins
  • Galectin 1
  • Galectin 3
  • Glycoproteins
  • Lectins
  • Plant Lectins
  • Polysaccharides
  • asialofetuin
  • Glycosyltransferases
  • N-Acetylgalactosaminyltransferases