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

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.