title: Effects of thyrotropin and thyrotropin-receptor-stimulating Graves' disease immunoglobulin G on cyclic adenosine monophosphate and hyaluronan production in nondifferentiated orbital fibroblasts of Graves' ophthalmopathy patients.
abstract: BACKGROUND
  Orbital fibroblasts are involved in the pathogenesis of Graves' ophthalmopathy (GO) by producing hyaluronan (HA), synthesized by three types of hyaluronan synthases (HAS1, HAS2, and HAS3). Thyrotropin receptors (TSHR) expressed in orbital fibroblasts activate the cyclic adenosine monophosphate (cAMP) pathway. Only sparse data are available at present supporting a role for TSHR activation in the regulation of HA in GO orbital fibroblasts. We hypothesize that TSHR activation, via cAMP signaling, results in induction of HAS1-3 mRNA expression and HA production by nondifferentiated GO orbital fibroblasts.
  
  
  METHODS
  Cultured nondifferentiated orbital fibroblasts obtained during orbital decompression surgery from 15 GO patients were stimulated with recombinant human TSH (rhTSH), TSHR-stimulating Graves' disease immunoglobulin G (GD-IgG) or forskolin (FSK), or interleukin-1beta (IL-1beta).
  
  
  RESULTS
  FSK significantly stimulated cAMP production, HAS1 and HAS3 mRNA expression, and HA secretion in orbital fibroblasts. IL-1beta slightly induced cAMP production, but induced HAS mRNA expression of all three isoforms and HA secretion. In contrast, the effects of rhTSH and GD-IgG on cAMP were modest and absent, respectively, and on HAS mRNA and HA synthesis were completely absent.
  
  
  CONCLUSIONS
  The strong increase in cAMP synthesis by FSK in nondifferentiated GO orbital fibroblasts results in increased HA synthesis, but TSHR activation by rhTSH or GD-IgG does not result in altered HA synthesis. Our results do not support a predominant role for GD-IgGs in the accumulation of orbital glycosaminoglycans; cytokines like IL-1beta seem largely responsible for excessive glycosaminoglycan production by nondifferentiated orbital fibroblasts in early immunopathogenesis of GO.
authors.affiliation: Department of Endocrinology and Metabolism, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands. c.j.vanzeijl@amc.uva.nl
authors.forename: Clementine J J
authors.initials: CJ
authors.lastname: van Zeijl
authors.forename: Eric
authors.initials: E
authors.lastname: Fliers
authors.forename: Chris J
authors.initials: CJ
authors.lastname: van Koppen
authors.forename: Olga V
authors.initials: OV
authors.lastname: Surovtseva
authors.forename: Marcel E
authors.initials: ME
authors.lastname: de Gooyer
authors.forename: Maarten P
authors.initials: MP
authors.lastname: Mourits
authors.forename: Wilmar M
authors.initials: WM
authors.lastname: Wiersinga
authors.forename: André M M
authors.initials: AM
authors.lastname: Miltenburg
authors.forename: Anita
authors.initials: A
authors.lastname: Boelen
mesh_terms: D003994:Bucladesine / Q000494:pharmacology; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000096:biosynthesis*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D006111:Graves Disease / Q000276:immunology*; D049970:Graves Ophthalmopathy / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D006820:Hyaluronic Acid / Q000096:biosynthesis*; D007074:Immunoglobulin G / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000096:biosynthesis; D008156:Luciferases / Q000378:metabolism; D009915:Orbit / Q000166:cytology; D012313:RNA / Q000096:biosynthesis / Q000302:isolation & purification; D011989:Receptors, Thyrotropin / Q000276:immunology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013972:Thyrotropin / Q000627:therapeutic use*
chemical_list: D007074:Immunoglobulin G; D053583:Interleukin-1beta; D011989:Receptors, Thyrotropin; D005576:Colforsin; D012313:RNA; D003994:Bucladesine; D013972:Thyrotropin; D006820:Hyaluronic Acid; D000242:Cyclic AMP; D008156:Luciferases