One of two chondrocyte-expressed isoforms of cartilage intermediate-layer protein functions as an insulin-like growth factor 1 antagonist

Arthritis Rheum. 2003 May;48(5):1302-14. doi: 10.1002/art.10927.

Abstract

Objective: Aging and osteoarthritic (OA) cartilage commonly demonstrate enhanced expression of the large, transforming growth factor beta (TGFbeta)-inducible glycoprotein cartilage intermediate-layer protein (CILP) as well as enhanced extracellular inorganic pyrophosphate (PPi) that promotes the deposition of calcium pyrophosphate dihydrate crystals. In normal chondrocytes, TGFbeta induces elevated chondrocyte extracellular PPi. Insulin-like growth factor 1 (IGF-1) normally blocks this response and reduces extracellular PPi. However, chondrocyte resistance to IGF-1 is observed in OA and aging. Because CILP was reported to chromatographically fractionate with PPi-generating nucleotide pyrophosphatase phosphodiesterase (NPP) activity, it has been broadly assumed that CILP itself has NPP activity. Our objective was to directly define CILP functions and their relationship to IGF-1 in chondrocytes.

Methods: Using primary cultures of articular chondrocytes from the knee, we defined the function of the previously described CILP (CILP-1) and of a recently described 50.6% identical protein that we designated the CILP-2 isoform.

Results: Both CILP isoforms were constitutively expressed by primary cultured articular chondrocytes, but only CILP-1 expression was detectable in cultured knee meniscal cartilage cells. Neither CILP isoform had intrinsic NPP activity. But CILP-1 blocked the ability of IGF-1 to decrease extracellular PPi, an activity specific for the CILP-1 N-terminal domain. The CILP-1 N-terminal domain also suppressed IGF-1-induced (but not TGFbeta-induced) proliferation and sulfated proteoglycan synthesis, and it inhibited ligand-induced IGF-1 receptor autophosphorylation.

Conclusion: Two CILP isoforms are differentially expressed by chondrocytes. Neither CILP isoform exhibits PPi-generating NPP activity. But, increased expression of CILP-1, via N-terminal domain-mediated inhibitory effects of CILP-1 on chondrocyte IGF-1 responsiveness, could impair chondrocyte growth and matrix repair and indirectly promote PPi supersaturation in aging and OA cartilage.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alkaline Phosphatase / metabolism
  • Animals
  • Cartilage, Articular / cytology
  • Cartilage, Articular / metabolism*
  • Cattle
  • Cells, Cultured
  • Chondrocytes / cytology
  • Chondrocytes / metabolism*
  • DNA, Complementary / analysis
  • Diphosphates / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Extracellular Matrix Proteins / metabolism*
  • Growth Substances / physiology*
  • Humans
  • Intercellular Signaling Peptides and Proteins / physiology*
  • Mice
  • Mice, Knockout
  • Phosphoric Diester Hydrolases / metabolism
  • Protein Isoforms
  • Pyrophosphatases / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Stifle
  • Transfection

Substances

  • DNA, Complementary
  • Diphosphates
  • Extracellular Matrix Proteins
  • Growth Substances
  • Intercellular Signaling Peptides and Proteins
  • Protein Isoforms
  • myotrophin
  • Alkaline Phosphatase
  • Phosphoric Diester Hydrolases
  • CILP protein, human
  • Pyrophosphatases
  • nucleotide pyrophosphatase - phosphodiesterase I