EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy

Graves’ ophthalmopathy (GO) is an extra-thyroidal complication of Graves’ disease which can lead to vision loss in severe cases. Currently, treatments of GO are not sufficiently effective, so novel therapeutic strategies are needed. As platelet-derived growth factor (PDGF)-BB induces several effector mechanisms in GO orbital fibroblasts including cytokine production and myofibroblast activation, this study aims to investigate the roles of histone lysine methyltransferases (HKMTs) in PDGF-BB-activated GO orbital fibroblasts by screening with HKMTs inhibitors library. From the total of twelve selective HKMT inhibitors in the library, EZH2, G9a and DOT1L inhibitors, DZNeP, BIX01294 and Pinometostat, respectively, prevented PDGF-BB-induced proliferation and hyaluronan production by GO orbital fibroblasts. However, only EZH2 inhibitor, DZNeP, significantly blocked pro-inflammatory cytokine production. For the HKMTs expression in GO orbital fibroblasts, PDGF-BB significantly and time-dependently induced EZH2, G9a and DOT1L mRNA expression. To confirm the role of EZH2 in PDGF-BB-induced orbital fibroblast activation, EZH2 silencing experiments revealed suppression of PDGF-BB-induced collagen type I and α-SMA expression along with decreasing histone H3 lysine 27 trimethylation (H3K27me3) level. In a more clinically relevant model than orbital fibroblast culture experiments, DZNeP treated GO orbital tissues significantly reduced pro-inflammatory cytokine production while slightly reduced ACTA2 mRNA expression. Our data is the first to demonstrate that among all HKMTs EZH2 dominantly involved in the expression of myofibroblast markers in PDGF-BB-activated orbital fibroblast from GO presumably via H3K27me3. Thus, EZH2 may represent a novel therapeutics target for GO.</p

EZH2 as a major histone methyltransferase in PDGF-BB-activated orbital fibroblast in the pathogenesis of Graves’ ophthalmopathy

Graves’ ophthalmopathy (GO) is an extra-thyroidal complication of Graves’ disease which can lead to vision loss in severe cases. Currently, treatments of GO are not sufficiently effective, so novel therapeutic strategies are needed. As platelet-derived growth factor (PDGF)-BB induces several effector mechanisms in GO orbital fibroblasts including cytokine production and myofibroblast activation, this study aims to investigate the roles of histone lysine methyltransferases (HKMTs) in PDGF-BB-activated GO orbital fibroblasts by screening with HKMTs inhibitors library. From the total of twelve selective HKMT inhibitors in the library, EZH2, G9a and DOT1L inhibitors, DZNeP, BIX01294 and Pinometostat, respectively, prevented PDGF-BB-induced proliferation and hyaluronan production by GO orbital fibroblasts. However, only EZH2 inhibitor, DZNeP, significantly blocked pro-inflammatory cytokine production. For the HKMTs expression in GO orbital fibroblasts, PDGF-BB significantly and time-dependently induced EZH2, G9a and DOT1L mRNA expression. To confirm the role of EZH2 in PDGF-BB-induced orbital fibroblast activation, EZH2 silencing experiments revealed suppression of PDGF-BB-induced collagen type I and α-SMA expression along with decreasing histone H3 lysine 27 trimethylation (H3K27me3) level. In a more clinically relevant model than orbital fibroblast culture experiments, DZNeP treated GO orbital tissues significantly reduced pro-inflammatory cytokine production while slightly reduced ACTA2 mRNA expression. Our data is the first to demonstrate that among all HKMTs EZH2 dominantly involved in the expression of myofibroblast markers in PDGF-BB-activated orbital fibroblast from GO presumably via H3K27me3. Thus, EZH2 may represent a novel therapeutics target for GO.</p

Skin tightening by an insulated microneedle radiofrequency device combined with lower blepharoplasty: a randomized fellow eye comparison study

Background : : Aging in the lower eyelids creates bags under the eyes from the protruding orbital fat, and periorbital wrinkles. Therefore, lower blepharoplasty should be combined with a skin tightening procedure to enhance the results.Objective: To compare the effects between lower blepharoplasty combined with an insulated microneedle radiofrequency (RF) device and lower blepharoplasty alone.Methods : : Twelve Asian (24 eyes) patients with baggy eyelids were included. One eye per patient was randomized for lower blepharoplasty combined with insulated microneedle RF devices (RF group), and the other eye for lower blepharoplasty alone (LB group). All patients underwent transconjunctival lower blepharoplasty with fat transposition and orbicularis oculi suspension. The primary outcome was the difference between both groups on the five-point Wrinkle Assessment Scale (WAS), assessed at baseline and at 2 months post-surgery. The secondary outcome was patient satisfaction using the Global Aesthetic Improvement Scale (GAIS).Results : : The WAS improvements were 75.0% (9/12 eyes) and 58.3% (7/12 eyes) in the RF and LB groups, respectively. The GAIS showed high patient satisfaction in both groups, with an average of 4.12 (3-5) in the RF group and 4.0 (2-5) in the LB group (p=0.157). There was no difference in the postoperative inflammation between both eyes at 1 week; and no decrease in the WAS score at 2 months or any adverse effects at 6 months were observed.Conclusion : : Wrinkle improvement was observed post-lower blepharoplasty with or without microneedle RF treatment. The insulated microneedle RF device proved safe when performed concurrently with lower blepharoplasty