title: Independent adipogenic and contractile properties of fibroblasts in Graves' orbitopathy: an in vitro model for the evaluation of treatments.
abstract: Graves' orbitopathy (GO) is a disfiguring and sometimes blinding disease, characterised by inflammation and swelling of orbital tissues, with fibrosis and adipogenesis being predominant features. Little is known about the disease aetiology and the molecular mechanisms driving the phenotypic changes in orbital fibroblasts are unknown. Using fibroblasts isolated from the orbital fat of undiseased individuals or GO patients, we have established a novel in vitro model to evaluate the dual profile of GO cells in a three-dimensional collagen matrix; this pseudo-physiological 3D environment allows measurement of their contractile and adipogenic properties. GO cells contracted collagen matrices more efficiently than control cells following serum or TGFβ1 stimulation, and showed a slightly increased ability to proliferate in the 3D matrix, in accordance with a fibro-proliferative phenotype. GO cells, unlike controls, also spontaneously differentiated into adipocytes in 3D cultures - confirming an intrinsic adipogenic profile. However, both control and GO cells underwent adipogenesis when cultured under pathological pressure levels. We further demonstrate that a Thy-1-low population of GO cells underlies the adipogenic - but not the contractile - phenotype and, using inhibitors, confirm that the contractile and adipogenic phenotypes are regulated by separate pathways. In view of the current lack of suitable treatment for GO, we propose that this new model testing the duality of the GO phenotype could be useful as a preclinical evaluation for the efficacy of potential treatments.
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: He
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: Caroline
authors.initials: C
authors.lastname: Fitchett
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Kozdon
authors.affiliation: Department of Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: Hari
authors.initials: H
authors.lastname: Jayaram
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom; Moorfields Eye Hospital, and the National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, UCL Partners AHSC, London, United Kingdom.
authors.forename: Geoffrey E
authors.initials: GE
authors.lastname: Rose
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom.
authors.forename: Maryse
authors.initials: M
authors.lastname: Bailly
authors.affiliation: Department of Cell Biology, UCL Institute of Ophthalmology, London, United Kingdom; Moorfields Eye Hospital, and the National Institute for Health Research (NIHR) Biomedical Research Centre at Moorfields Eye Hospital NHS Foundation Trust and UCL Institute of Ophthalmology, UCL Partners AHSC, London, United Kingdom.
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Ezra
mesh_terms: D050156:Adipogenesis; D000368:Aged; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000473:pathology*; D049970:Graves Ophthalmopathy / Q000473:pathology*; D006801:Humans; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D010641:Phenotype; D011312:Pressure; D018800:Thy-1 Antigens / Q000378:metabolism
chemical_list: D018800:Thy-1 Antigens
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