Link N is cleaved by human annulus fibrosus cells generating a fragment with retained biological activity

J Orthop Res. 2014 Sep;32(9):1189-97. doi: 10.1002/jor.22653. Epub 2014 May 23.

Abstract

Presently, there are no established treatments to prevent, stop or even retard back pain arising from disc degeneration. Previous studies have shown that Link N can act as a growth factor and stimulate the synthesis of proteoglycans and collagens, in IVD. However, the sequences in Link N involved in modulating cellular activity are not well understood. To determine if disc cells can proteolytically process Link N, human disc cells were exposed to native Link N over a 48 h period and mass spectrometric analysis revealed that a peptide spanning residues 1-8 was generated in the presence of AF cells but not NP cells. Link N 1-8 significantly induced proteoglycan production in the presence of IL-1β NP and AF cells, confirming that the biological effect is maintained in the first 8 amino acids of the peptide and indicating that the effect is sustained in an inflammatory environment. Thus Link-N 1-8 could be a promising candidate for biologically induced disc repair, and the identification of such a stable specific peptide may facilitate the design of compounds to promote disc repair and provide alternatives to surgical intervention for early stage disc degeneration.

Keywords: IVD degeneration and regeneration; Link N; biological repair; organ culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Amino Acid Sequence
  • Animals
  • Biological Factors / metabolism*
  • Biological Factors / pharmacology*
  • Cattle
  • Cells, Cultured
  • Collagen / metabolism
  • Female
  • Humans
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Intercellular Signaling Peptides and Proteins / pharmacology
  • Intervertebral Disc / drug effects*
  • Intervertebral Disc / metabolism*
  • Intervertebral Disc / pathology
  • Intervertebral Disc Degeneration / metabolism
  • Intervertebral Disc Degeneration / pathology
  • Lumbar Vertebrae / pathology
  • Male
  • Middle Aged
  • Molecular Sequence Data
  • Peptide Fragments / metabolism*
  • Peptide Fragments / pharmacology*
  • Proteoglycans / metabolism
  • Regeneration / drug effects
  • Thoracic Vertebrae / pathology
  • Young Adult

Substances

  • Biological Factors
  • Intercellular Signaling Peptides and Proteins
  • Peptide Fragments
  • Proteoglycans
  • Collagen