title: Orbital fibroblasts from thyroid eye disease patients differ in proliferative and adipogenic responses depending on disease subtype.
abstract: OBJECTIVE
  Thyroid eye disease (TED) patients are classified as type I (predominantly fat compartment enlargement) or type II (predominantly extraocular muscle enlargement) based on orbital imaging. Orbital fibroblasts (OFs) can be driven to proliferate or differentiate into adipocytes in vitro. We tested the hypothesis that type I OFs undergo more adipogenesis than type II OFs, whereas type II OFs proliferate more than type I OFs. We also examined the effect of cyclooxygenase (COX) inhibitors on OF adipogenesis and proliferation.
  
  
  METHODS
  Type I, type II, and non-TED OFs were treated with transforming growth factor-beta (TGFβ) to induce proliferation and with 15-deoxy-Δ(-12,14)-prostaglandin J2 (15d-PGJ2) to induce adipogenesis. Proliferation was measured using the [(3)H]thymidine assay, and adipogenesis was measured using the AdipoRed assay, Oil Red O staining, and flow cytometry. The effect of COX inhibition on adipogenesis and proliferation was also studied.
  
  
  RESULTS
  Type II OFs incorporated 1.7-fold more [(3)H]thymidine than type I OFs (P < 0.05). Type I OFs accumulated 4.8-fold more lipid than type II OFs (P < 0.05) and 12.6-fold more lipid than non-TED OFs (P < 0.05). Oil Red O staining and flow cytometry also demonstrated increased adipogenesis in type I OFs compared to type II and non-TED OFs. Cyclooxygenase inhibition significantly decreased proliferation and adipogenesis in type II OFs, but not type I OFs.
  
  
  CONCLUSIONS
  We have demonstrated that OFs from TED patients have heterogeneous responses to proproliferative and proadipogenic stimulators in vitro in a manner that corresponds to their different clinical manifestations. Furthermore, we demonstrated a differential effect of COX inhibitors on type I and type II OF proliferation and adipogenesis.
authors.affiliation: Flaum Eye Institute, University of Rochester School of Medicine and Dentistry, Rochester, New York.
authors.forename: Ajay E
authors.initials: AE
authors.lastname: Kuriyan
authors.forename: Collynn F
authors.initials: CF
authors.lastname: Woeller
authors.forename: Charles W
authors.initials: CW
authors.lastname: O'Loughlin
authors.forename: Richard P
authors.initials: RP
authors.lastname: Phipps
authors.forename: Steven E
authors.initials: SE
authors.lastname: Feldon
mesh_terms: D050156:Adipogenesis / Q000187:drug effects*; D000704:Analysis of Variance; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D049109:Cell Proliferation / Q000187:drug effects*; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000187:drug effects*; D005434:Flow Cytometry; D049970:Graves Ophthalmopathy / Q000188:drug therapy / Q000473:pathology*; D006801:Humans; D009915:Orbit / Q000473:pathology*; D015230:Prostaglandin D2 / Q000031:analogs & derivatives / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000494:pharmacology
chemical_list: C477819:15-deoxyprostaglandin J2; D016861:Cyclooxygenase Inhibitors; D016212:Transforming Growth Factor beta; D015230:Prostaglandin D2
keywords: adipogenesis; orbital fibroblasts; proliferation; thyroid eye disease