Effect of tryptase inhibition on joint inflammation: a pharmacological and lentivirus-mediated gene transfer study

Arthritis Res Ther. 2017 Jun 6;19(1):124. doi: 10.1186/s13075-017-1326-9.

Abstract

Background: Increasing evidences indicate that an unbalance between tryptases and their endogenous inhibitors, leading to an increased proteolytic activity, is implicated in the pathophysiology of rheumatoid arthritis. The aim of the present study was to evaluate the impact of tryptase inhibition on experimental arthritis.

Methods: Analysis of gene expression and regulation in the mouse knee joint was performed by RT-qPCR and in situ hybridization. Arthritis was induced in male C57BL/6 mice with mBSA/IL-1β. Tryptase was inhibited by two approaches: a lentivirus-mediated heterologous expression of the human endogenous tryptase inhibitor, sperm-associated antigen 11B isoform C (hSPAG11B/C), or a chronic treatment with the synthetic tryptase inhibitor APC366. Several inflammatory parameters were evaluated, such as oedema formation, histopathology, production of IL-1β, -6, -17A and CXCL1/KC, myeloperoxidase and tryptase-like activities.

Results: Spag11c was constitutively expressed in chondrocytes and cells from the synovial membrane in mice, but its expression did not change 7 days after the induction of arthritis, while tryptase expression and activity were upregulated. The intra-articular transduction of animals with the lentivirus phSPAG11B/C or the treatment with APC366 inhibited the increase of tryptase-like activity, the late phase of oedema formation, the production of IL-6 and CXCL1/KC. In contrast, neutrophil infiltration, degeneration of hyaline cartilage and erosion of subchondral bone were not affected.

Conclusions: Tryptase inhibition was effective in inhibiting some inflammatory parameters associated to mBSA/IL-1β-induced arthritis, notably late phase oedema formation and IL-6 production, but not neutrophil infiltration and joint degeneration. These results suggest that the therapeutic application of tryptase inhibitors to rheumatoid arthritis would be restrained to palliative care, but not as disease-modifying drugs. Finally, this study highlighted lentivirus-based gene delivery as an instrumental tool to study the relevance of target genes in synovial joint physiology and disease.

Keywords: APC366; Inflammation; Protease inhibitor; Rheumatoid arthritis; SPAG11B; Synovial joint; Tryptase; β-defensin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigens, Surface / genetics
  • Antigens, Surface / metabolism
  • Arthritis, Experimental / genetics
  • Arthritis, Experimental / metabolism
  • Arthritis, Experimental / therapy
  • Arthritis, Rheumatoid / genetics
  • Arthritis, Rheumatoid / metabolism
  • Arthritis, Rheumatoid / therapy
  • Chondrocytes / metabolism
  • Cytokines / metabolism
  • Dipeptides / pharmacology
  • Gene Transfer Techniques*
  • HEK293 Cells
  • Humans
  • Inflammation / genetics
  • Inflammation / metabolism*
  • Inflammation / therapy
  • Knee Joint / drug effects
  • Knee Joint / metabolism*
  • Knee Joint / pathology
  • Lentivirus / genetics
  • Male
  • Mice, Inbred C57BL
  • Synovial Membrane / metabolism
  • Tryptases / antagonists & inhibitors
  • Tryptases / genetics
  • Tryptases / metabolism*

Substances

  • Antigens, Surface
  • Cytokines
  • Dipeptides
  • N-(1-hydroxy-2-naphthoyl)arginyl-prolinamide
  • SPAG11B protein, human
  • Tryptases