Autoantibodies in Melanoma-Associated Retinopathy Recognize an Epitope Conserved Between TRPM1 and TRPM3

Invest Ophthalmol Vis Sci. 2017 May 1;58(5):2732-2738. doi: 10.1167/iovs.17-21443.

Abstract

Purpose: Melanoma-associated retinopathy (MAR) is a paraneoplastic syndrome associated with malignant melanoma and the presence of anti-retinal autoantibodies, including autoantibodies against transient receptor potential melanopsin 1 (TRPM1), a cation channel expressed by both melanocytes and retinal bipolar cells. The goal of this study was to further map the antigenic epitope.

Methods: Patient sera were tested by immunofluorescence and Western blotting on HEK293 cells transfected with enhanced green fluorescent protein (EGFP)-TRPM1 fusion constructs and mouse retina sections.

Results: The epitope recognized by MAR patient sera was mapped to a region encoded by exons 9 and 10 of the human TRPM1 gene. This region of TRPM1 is highly conserved with TRPM3, and indeed MAR sera were found to cross-react with TRPM3, a closely related channel expressed in the retinal pigment epithelium (RPE).

Conclusions: These results indicate that TRPM1 autoantibodies in MAR patient sera recognize a short, intracellular segment of TRPM1. Cross-reactivity with TRPM3 in the RPE may account for other visual symptoms that are experienced by some MAR patients such as retinal and RPE detachments. We propose that TRPM1 autoantibodies are generated in response to abnormal TRPM1 polypeptides encoded by an alternate mRNA splice variant expressed by malignant melanocytes.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Autoantibodies / blood*
  • Blotting, Western
  • Cross Reactions
  • Epitopes*
  • Exons / genetics
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Green Fluorescent Proteins
  • HEK293 Cells
  • Humans
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Paraneoplastic Syndromes, Ocular / immunology*
  • Recombinant Fusion Proteins / genetics
  • TRPM Cation Channels / genetics
  • TRPM Cation Channels / immunology*
  • Transfection

Substances

  • Autoantibodies
  • Epitopes
  • Recombinant Fusion Proteins
  • TRPM Cation Channels
  • TRPM1 protein, human
  • TRPM3 protein, human
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins