Skip to main content
Dryad logo

Data from: Novel roles of chloroquine and hydroxychloroquine in Graves’ orbitopathy therapy by targeting orbital fibroblasts

Citation

Guo, Yan et al. (2020), Data from: Novel roles of chloroquine and hydroxychloroquine in Graves’ orbitopathy therapy by targeting orbital fibroblasts, Dryad, Dataset, https://doi.org/10.5061/dryad.5tb2rbp1k

Abstract

Context: Graves’ orbitopathy (GO) causes infiltrative exophthalmos by inducing excessive proliferation, adipogenesis and glycosaminoglycan production in orbital fibroblasts (OFs). Interference with OF autophagy is a potential therapy for proptosis.

Objectives: Here, we aimed to evaluate the effects of chloroquine (CQ) and hydroxychloroquine (HCQ), the autophagy inhibitors commonly used in clinical practice, on OFs.

Design/Setting/Participants: OFs isolated from patients with GO (GO-OFs) or control persons (non-GO-OFs) were cultured in proliferation medium (PM) or subjected to differentiation medium (DM). OFs were treated with CQ or HCQ (0, 2, 5,10 μM), and subsequently examined in vitro.

Main Outcome Measures: CCK-8, EdU incorporation and flow cytometry assays were used to assess cellular viability. Adipogenesis was assessed with Western blot analysis, real-time PCR, and Oil Red O staining. Hyaluronan production was determined by real-time PCR and ELISA. Autophagy flux was detected using RFP-GFP-LC3 fluorescent staining and Western blot analysis.

Results: CQ/HCQ halted proliferation and adipogenesis in GO-OFs in a concentration-dependent manner through blockage of autophagy, which were not detected in non-GO-OFs. Besides, inhibitory effect of CQ/HCQ on hyaluronan secretion of GO-OFs was also concentration-dependent, mediated by downregulation of hyaluronan synthase 2 (HAS2) rather than hyaluronidases. Moreover, CQ (10 mM) induced GO-OFs apoptosis without aggravating oxidative stress.

Conclusions: The antimalarials CQ/HCQ affect proliferation, adipogenesis and hyaluronan generation in GO-OFs by inhibiting autophagy, providing proof of concept that they can treat GO as autophagy inhibitors.

Usage Notes

Supplemental figures 1-3

Funding

National Key Research and Development Program of China, Award: 2018YFC1314100

National Natural Science Foundation of China, Award: 818,705,578,157,262,000,000,000

Natural Science Foundation of Guangdong Province, Award: 2016A030310169

Key Field Research and Development Program of Guangdong Province, Award: 2019B020230001

Graduate Innovative Development Program, Award: 19ykyjs08