Supplementary data from: 2'-O-Galloylhyperin prevents tissue remodeling in thyroid eye disease: prospects as a thyrotropin receptor antagonist
Creators: name: Guo, Yan
name: Liu, Boyuan
name: Liang, Weiwei
name: Cheng, Yanglei
name: Li, Hai
name: Guan, Hongyu
name: Xiao, Haipeng
name: Li, Yanbing
Publisher: name: Zenodo
Year: 2024
Subjects: thyroid eye disease
tissue remodeling
TSHR
antagonist
Organoids

description: Context: Thyroid eye disease (TED) is a challenging condition owing to relentless orbital tissue remodeling, with thyroid-stimulating hormone receptor (TSHR) in orbital fibroblasts (OFs) serving as a promising therapeutic target.

Objectives: This study seeks to discover potential TSHR inhibitors among FDA-approved drugs and evaluate their effects on TED-OFs.

Design/Participants: Adipose tissues were sourced from the patients with or without TED. Isolated OFs were cultivated in proliferation medium or stimulated for adipogenic/fibrotic differentiation in 2D/3D models, treated by 2'-O-GH (0, 5, 20, and 50μM).

Main Outcome Measures: Using structure-based virtual screening (SBVS), potential TSHR antagonists were identified. Cellular proliferation was analyzed by EdU incorporation, flow cytometry and spheroid size. Adipogenesis was determined by Oil Red O staining, Western blot and immunofluorescence. Fibrosis was assessed using woundhealing assays, Western blot and immunofluorescence. Cyclic adenosine monophosphate (cAMP), hyaluronan and cytokine were quantified by ELISA.

Results: Herein, the FDA-approved drug 2'-O-Galloylhyperin (2'-O-GH) dose-dependently decreased cAMP production and the subsequent CREB phosphorylation stimulated by a TSHR stimulating monoclonal autoantibody M22, which was reversed by a consistently activated mutation of TSHR (L629F). As expected, 2'-O-GH attenuated lipid accumulation in TED-OFs, along with down-regulation of key adipogenic markers. And 2'-O-GH ameliorated hyaluronan production during adipogenesis. Notably, 2'-O-GH dampened wound closure and fibrotic differentiation of TED-OF stimualted by TGF-β1. Correspondingly, proliferation of TED-OFs was halted by 2'-O-GH.

Conclusions: 2'-O-GH promises to prevent the tissue remodeling of TED by exerting inhibitory effects on proliferation, differentiation and hyaluronan deposition by inhibiting TSHR activation, implying its potential therapeutic value for TED.
description: Funding provided by: National Natural Science Foundation of ChinaROR ID: https://ror.org/01h0zpd94Award Number: 82200876
Funding provided by: National Natural Science Foundation of ChinaROR ID: https://ror.org/01h0zpd94Award Number: 81870557
Funding provided by: National Natural Science Foundation of ChinaROR ID: https://ror.org/01h0zpd94Award Number: 81572624
Funding provided by: Natural Science Foundation of Guangdong ProvinceCrossref Funder Registry ID: Award Number: 2024A1515013290
Funding provided by: Key Field Research and Development Program of Guangdong Province, ChinaCrossref Funder Registry ID: Award Number: 2019B020230001
Funding provided by: Guangdong Basic and Applied Basic Research FoundationCrossref Funder Registry ID: Award Number: 2020A1515010049
Funding provided by: Sun Yat-sen UniversityROR ID: https://ror.org/0064kty71Award Number: 24qnpy346