Objectives A beneficial effect of sirolimus in Graves’ orbitopathy (GO) was reported, suggesting a possible use in clinical practice. We conducted an observational, single-centre, no-profit, clinical study to investigate the efficacy of sirolimus as a second-line treatment for moderate-to-severe, active GO compared with methylprednisolone. Methods Data from consecutive patients given sirolimus (2 mg orally on first day, followed by 0.5 mg/day for 12 weeks) or methylprednisolone [500 mg iv/weekly (6 weeks), 250 mg/weekly (6 weeks)] as a second-line treatment were collected and compared. Primary objective: overall GO outcome at 24 weeks, based on a composite evaluation. Secondary objectives at 24 weeks: (1) improvement in quality of life, evaluated using a specific uestionnaire (GO-QoL); (2) reduction in proptosis; (3) reduction in the clinical activity score (CAS); (4) improvement of eye ductions; and (5) reduction in eyelid aperture. Results Data from 30 patients (15 per group) treated between January 15, 2020, and June 15, 2021, were analysed. Proportion of GO responders (primary outcome) at 24 weeks was significantly greater in sirolimus group compared with methylprednisolone group (86.6% vs 26.6%; OR: 17.8; 95% CI from 2.7 to 116.8; P = 0.0026). GO-quality of life (GO-QoL) score was greater in sirolimus group. Proportion of proptosis responders was greater in sirolimus group, as well as proportion of clinical activity score (CAS) responders. No serious adverse events were observed, with no differences between groups. Conclusions Sirolimus seems to be an effective second-line treatment for GO. Further randomized clinical trials are needed to confirm our observations.

Context A role of DNA methylation in Graves’ orbitopathy (GO) has been proposed. Design. To investigate DNA methylation and gene expression in orbital fibroblasts from control and GO patients, under basal conditions or following challenge with an anti-TSH receptor antibody (M22) or cytokines involved in GO; to investigate the relationship between DNA methylation and cell function (proliferation); to perform a methylome analysis. Setting. Referral Center. Materials. Orbital fibroblasts from six GO and six control patients. Intervention. None. Main Outcome Measure. Methylome analysis of the whole genome. Results. Global DNA methylation increased significantly both in control and GO fibroblasts upon incubation with M22. Expression of two selected genes (CYP19A1 and AIFM2) was variably affected by M22 and interleukin-6. M22 increased cell proliferation in control and GO fibroblasts, which correlated with global DNA methylation. Methylome analysis revealed 19,869 DNA regions differently methylated in GO fibroblasts, encompassing 3,957 genes and involving CpG islands, shores and shelves. One-hundred and nineteen gene families and subfamilies, 89 protein groups, 402 biological processes and seven pathways were involved. Three genes found to be differentially expressed were concordantly hyper- or hypomethylated. Among the differently methylated genes, insulin-like growth factor-1 receptor and several fibroblast growth factors and receptors were included. Conclusions We propose that, when exposed to an autoimmune environment, orbital fibroblasts undergo hyper- or hypomethylation of certain genes, involving CpG promoters, which results in differential gene expression, which may be responsible for functional alterations, in particular higher proliferation, and ultimately for the GO phenotype in vivo.

Objective A relationship between thyroid and non-organ-specific autoimmunity could be relevant for Graves’ orbitopathy (GO), which affects connective tissue. We investigated the association between GO and anti-nuclear antibodies (ANAs). Methods Retrospective investigation was conducted in 265 patients with Graves’ disease (GD), 158 with and 107 without GO. Primary outcome was: prevalence of ANAs in GO vs no-GO. Secondary outcomes were: (1) relationship between ANAs and GO features; (2) prevalence of ANAs in GD compared with non-autoimmune hyperthyroidism [(78 patients with toxic nodular goiter (TNG)]; (3) distribution of ANA patterns. Results ANAs were detected in 212 (80%) GD patients, but prevalence did not differ between GO (79.7%) and no-GO (80.3%). Higher ANA titers (1:160) were more common in GO (51.5 vs 38.3%), but only nearly significantly (OR 0.5; 95% CI: 0.3–1; P = 0.059). Proptosis was lower in ANA-positive patients (mean difference: − 1.4 mm; 95% CI from − 2.5 to − 0.3; P = 0.011), in whom nearly significantly lower CAS (Mann–Whitney U: 1.5; P = 0.077) and eyelid aperture (mean difference: − 0.9 mm; 95% CI from − 2 to 0; P = 0.062) were observed. Prevalence of ANAs in GD was lower than in TNG (80 vs 91%; OR 0.3; 95% CI: 0.1–0.9; P = 0.028), but nuclear speckled pattern was more frequent (OR 22.9; 95% CI 1.3–381.3; P = 0.028). Conclusions Although ANAs are not more frequent in GO, they seem to exert a protective role on its severity and on development of GD. A switch of T cell population in ANA-positive patients, resulting in a different phenotype, may be responsible. Further studies are needed to investigate the mechanisms.