title: Circulating microRNA predicts insensitivity to glucocorticoid therapy in Graves' ophthalmopathy.
abstract: Glucocorticoid (GC) insensitivity occurs commonly in Graves' ophthalmopathy (GO), and GC therapy is associated with major adverse effects. A reliable and easily accessible biomarker is required to predict the outcome of GC therapy. This study aimed to evaluate the performance of circulating microRNA (miRNA) to predict GC insensitivity in GO patients. A total of 35 consecutive patients were included in this study. A cumulative dose of 4.5 g of methylprednisolone (MP) was administered intravenously for 12 weeks. Pretreatment serum miRNAs from the best- (N = 5) and worst- (N = 4) responding patients were profiled using miScript PCR arrays and validated by quantitative PCR in all patients. We calculated the predictive value of pretreatment assays of serum miRNAs with regard to GC insensitivity. We further investigated the roles of target miRNAs in modulating NF-κB activity and restoring transrepression of an NF-κB reporter by dexamethasone. Nine miRNAs displayed significant differences between responsive and resistant patients by miScript PCR arrays. Validation of the top two miRNAs in all 35 patients confirmed a significantly lower serum level of miR-224-5p (p = 0.0048) in resistant patients. A multivariate logistic regression model identified a composite biomarker combining baseline serum miR-224-5p and TRAb was independently associated with GC response (OR: 2.565, 95 % CI 1.011-6.505, p = 0.047). Receiver operating characteristic (ROC) curves analysis revealed the composite marker combining miR-224-5p and TRAb led to a 91.67 % positive prediction value (PPV) and a 69.56 % negative prediction value (NPV) with regard to GC resistance. Overexpression of miR-224-5p restored transrepression of the NF-κB reporter by dexamethasone under induced resistance, which may be via targeting GSK-3β to increase GR protein level. Our study demonstrated baseline serum miR-224-5p was associated with GC sensitivity in GO and in vitro overexpression of miR-224-5p restored GC sensitivity in a resistant cell model. A parameter combined serum miR-224-5p and TRAb could effectively predict GC sensitivity in GO patients.
authors.affiliation: Shanghai Key Laboratory for Endocrine Tumors, Shanghai Clinical Center for Endocrine and Metabolic Diseases, Shanghai Institute of Endocrine and Metabolic Diseases and Shanghai E-institute for Endocrinology, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, 197 Ruijin 2nd Road, Shanghai, 200025, People's Republic of China.
authors.forename: Liyun
authors.initials: L
authors.lastname: Shen
authors.forename: Fengjiao
authors.initials: F
authors.lastname: Huang
authors.forename: Lei
authors.initials: L
authors.lastname: Ye
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
authors.forename: Xiaofang
authors.initials: X
authors.lastname: Zhang
authors.forename: Shu
authors.initials: S
authors.lastname: Wang
authors.forename: Weiqing
authors.initials: W
authors.lastname: Wang
authors.forename: Guang
authors.initials: G
authors.lastname: Ning
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000097:blood; D003907:Dexamethasone / Q000494:pharmacology*; D004351:Drug Resistance / Q000502:physiology*; D005260:Female; D005500:Follow-Up Studies; D005938:Glucocorticoids / Q000008:administration & dosage / Q000494:pharmacology*; D049970:Graves Ophthalmopathy / Q000097:blood* / Q000188:drug therapy*; D006801:Humans; D018828:Immunoglobulins, Thyroid-Stimulating / Q000097:blood*; D008297:Male; D008775:Methylprednisolone / Q000008:administration & dosage / Q000494:pharmacology*; D035683:MicroRNAs / Q000097:blood*; D008875:Middle Aged
chemical_list: D015415:Biomarkers; D005938:Glucocorticoids; D018828:Immunoglobulins, Thyroid-Stimulating; C546140:MIRN224 microRNA, human; D035683:MicroRNAs; C094372:thyrotropin-binding inhibitory immunoglobulin; D003907:Dexamethasone; D008775:Methylprednisolone