Label-free LC-MSMS analysis of vitreous from autoimmune uveitis reveals a significant decrease in secreted Wnt signalling inhibitors DKK3 and SFRP2

J Proteomics. 2012 Jul 19;75(14):4545-54. doi: 10.1016/j.jprot.2012.04.052. Epub 2012 May 23.

Abstract

Equine recurrent uveitis is a severe and frequent blinding disease in horses which presents with auto-reactive invading T-cells, resulting in the destruction of the inner eye. Infiltration of inflammatory cells into the retina and vitreous is driven by currently unknown guidance cues, however surgical removal of the vitreous (vitrectomy) has proven therapeutically successful. Therefore, proteomic analyses of vitrectomy samples are likely to result in detection of proteins contributing to disease pathogenesis. Vitreous from healthy and ERU diseased horses were directly compared by quantitative mass spectrometry based on label-free quantification of peak intensities across samples. We found a significant upregulation of complement and coagulation cascades and downregulation of negative paracrine regulators of canonical Wnt signalling including the Wnt signalling inhibitors DKK3 and SFRP2. Based on immunohistochemistry, both proteins are expressed in equine retina and suggest localisation to retinal Müller glial cells (RMG), which may be the source cells for these proteins. Furthermore, retinal expression levels and patterns of DKK3 change in response to ERU. Since many other regulated proteins identified here are associated with RMG cells, these cells qualify as the prime responders to autoimmune triggers.

MeSH terms

  • Animals
  • Autoimmune Diseases / metabolism
  • Autoimmune Diseases / veterinary*
  • Chromatography, Liquid / methods
  • Down-Regulation
  • Horse Diseases / metabolism*
  • Horses
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Mass Spectrometry / methods
  • Membrane Proteins / metabolism*
  • Signal Transduction
  • Staining and Labeling
  • Uveitis / metabolism
  • Uveitis / veterinary*
  • Vitreous Body
  • Wnt Proteins / metabolism*

Substances

  • Intercellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Wnt Proteins