title: Novel markers of inflammation identified in tumor necrosis factor receptor-associated periodic syndrome (TRAPS) by transcriptomic analysis of effects of TRAPS-associated tumor necrosis factor receptor type I mutations in an endothelial cell line.
abstract: OBJECTIVE
  To analyze the effects of tumor necrosis factor receptor-associated periodic syndrome (TRAPS)-associated mutant tumor necrosis factor receptor type I (TNFRI) expression in a cell type directly relevant to the inflammation in TRAPS, and to identify novel markers associated with mutant TNFRI expression.
  
  
  METHODS
  Transcriptome analysis on 30,000 human genes was performed on SK-Hep-1 human endothelial cells transfected with either wild-type (WT) or TRAPS-associated mutant TNFRI. Quantitative reverse transcriptase-polymerase chain reaction and protein expression levels measured by enzyme-linked immunosorbent assay verified transcriptional changes for selected genes both in supernatants from cells expressing mutant TNFRI and in patient plasma.
  
  
  RESULTS
  Cells expressing mutant TNFRI showed up-regulation of multiple proinflammatory genes relative to WT transfectants, including genes for pentraxin 3, granulocyte-macrophage colony-stimulating factor, granulocyte colony-stimulating factor, CCL2, and CCL5, which were also expressed as proteins. In addition, the expression of most of these markers was increased in the plasma and peripheral blood mononuclear cells from TRAPS patients relative to those from healthy controls. The cysteine mutations (C33Y and C52F), which are associated with a more severe clinical phenotype, induced more genes than the low-penetrance mutation R92Q, which is associated with a milder phenotype. The expression of most genes was induced by a death domain (DD)-dependent mechanism, since they were not induced by expression of TNFRI mutants with an inactivated DD.
  
  
  CONCLUSIONS
  TRAPS-associated TNFRI mutants induce the expression of multiple genes encoding inflammatory molecules, cellular receptors, transcription factors, and regulators of apoptosis in endothelial cells that require the cytoplasmic signaling properties of the receptor. Different mutants have specific expression profiles, indicating mutation-specific effects. The expression of some of these markers was also elevated in samples from TRAPS patients.
authors.affiliation: Institute of Infection, Immunity and Inflammation, School of Molecular Medical Sciences, University of Nottingham, and Queen's Medical Centre, Nottingham, UK.
authors.forename: Susana L
authors.initials: SL
authors.lastname: Rebelo
authors.forename: Mohammad R
authors.initials: MR
authors.lastname: Amel-Kashipaz
authors.forename: Paul M
authors.initials: PM
authors.lastname: Radford
authors.forename: Susan E
authors.initials: SE
authors.lastname: Bainbridge
authors.forename: Roel
authors.initials: R
authors.lastname: Fiets
authors.forename: Johnny
authors.initials: J
authors.lastname: Fang
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: McDermott
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Powell
authors.forename: Ian
authors.initials: I
authors.lastname: Todd
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Tighe
mesh_terms: D001327:Autoimmune Diseases / Q000235:genetics / Q000276:immunology; D015415:Biomarkers; D006528:Carcinoma, Hepatocellular; D045744:Cell Line, Tumor; D004727:Endothelium / Q000166:cytology; D010505:Familial Mediterranean Fever / Q000235:genetics* / Q000276:immunology*; D020869:Gene Expression Profiling; D006801:Humans; D008113:Liver Neoplasms; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards; D009942:Organometallic Compounds; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics* / Q000276:immunology*; D015203:Reproducibility of Results; D014162:Transfection
chemical_list: D015415:Biomarkers; D009942:Organometallic Compounds; C524943:RT-03 compound; D047888:Receptors, Tumor Necrosis Factor, Type I