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Circulating microRNA predicts insensitivity to glucocorticoid therapy in Graves’ ophthalmopathy

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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.

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Acknowledgments

This study was supported by the grants from Science and Technology Commission of Shanghai Municipality [11DJ1400200, 13QH1402000] and China National Natural Science Foundation [30900701, 81200569]. The authors are grateful to the ophthalmologist Dr. Qin Jiao (Ruijin Hospital, Shanghai Jiaotong University, School of Medicine) who evaluated patients’ conditions. The authors also acknowledge Dr. Qinyun Ma, Dr. Xuelian Xiong and Dr. Mingwei Li for the kind gifts of luciferase reporters. And we thank Dr. Tiange Wang (Ruijin Hospital, Shanghai Jiaotong University, School of Medicine) for providing statistical assistance.

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The authors have nothing to declare.

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    Authors and Affiliations

    1. 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

      Liyun Shen, Fengjiao Huang, Lei Ye, Wei Zhu, Xiaofang Zhang, Shu Wang, Weiqing Wang & Guang Ning

    2. Laboratory for Endocrine & Metabolic Diseases of Institute of Health Science, Shanghai Jiaotong University School of Medicine and Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 227 South Chong Qing Road, Shanghai, 200025, People’s Republic of China

      Guang Ning

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    1. Liyun Shen
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    Correspondence to Lei Ye or Weiqing Wang.

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    Liyun Shen and Fengjiao Huang have contributed equally to this work.

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    Shen, L., Huang, F., Ye, L. et al. Circulating microRNA predicts insensitivity to glucocorticoid therapy in Graves’ ophthalmopathy. Endocrine 49, 445–456 (2015). https://doi.org/10.1007/s12020-014-0487-4

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