Department of Ophthalmology, Eye & ENT Hospital, Shanghai Medical College, Fudan University, 83 Fenyang Road, Shanghai, China; NHC Key Laboratory of Myopia, Fudan University, 83 Fenyang Road, Shanghai, China; Key Laboratory of Myopia, Ministry of Health, Fudan University, 83 Fenyang Road, Shanghai, China.
Department of Ophthalmology, Changzheng Hospital, Second Military Medical University, 415 Fengyang Road, Shanghai, China.
Exp Eye Res. 2020 Jul;196:108036. doi: 10.1016/j.exer.2020.108036. Epub 2020 May 4.
Our study aimed to investigate the differentially expressed circRNAs and their potential roles in orbital adipose/connective tissue from patients with thyroid-associated ophthalmopathy (TAO). The orbital adipose/connective tissue samples from three TAO patients and three control individuals were collected for RNA sequencing after depletion of ribosomal RNA. Differentially expressed mRNAs and up-regulated circRNAs were used for co-expression analysis. Functional and pathway enrichment analysis were conducted for the up- and down-regulated mRNAs in the circRNA-mRNA co-expression network. Meanwhile, circRNA-miRNA interaction network was established by miRanda software. The expression levels of mRNAs and circRNAs in control and TAO samples were determined by qRT-PCR. Among all the 16,329 circRNAs predicted from RNA sequencing data, 163 circRNAs (95 down-regulated and 68 up-regulated) were differentially expressed in TAO samples. Besides, 607 differentially expressed mRNAs were identified. The co-expression analysis showed circRNA_14940 was correlated with CCND1 and TNXB, while circRNA_10135 was correlated with PTGFR, and circRNA_14936 was correlated with TNFRSF19. The up-regulated CCND1 participated in Wnt signaling pathway. The down-regulated TNXB was involved in the ECM-receptor interaction, focal adhesion, and PI3K-Akt signaling pathway. PTGFR participated in neuroactive ligand-receptor interaction and calcium signaling pathway. TNFRSF19 was involved in cytokine-cytokine receptor interaction. In the interaction network, circRNA_14936 could interact with hsa-miR-10392-3p, and circRNA_12367 could interact with hsa-miR-1228-3p. Moreover, the expression changes of MMP2, TNXB, PTGFR, CCND1, and TNFRSF19, as well as circRNA_14936, circRNA_14940, and circRNA_12367 were validated by qRT-PCR. In conclusion, the differentially expressed circRNAs might participate in pathogenesis of TAO, and we speculated that circRNA_14940-CCND1-Wnt signaling pathway might be an important regulatory axis.
我们的研究旨在探讨甲状腺相关眼病（TAO）患者眼眶脂肪/结缔组织中差异表达的 circRNA 及其潜在作用。从 3 名 TAO 患者和 3 名对照个体的眼眶脂肪/结缔组织样本中，在去除核糖体 RNA 后，进行 RNA 测序。差异表达的 mRNA 和上调的 circRNA 用于共表达分析。对 circRNA-mRNA 共表达网络中的上调和下调 mRNA 进行功能和通路富集分析。同时，通过 miRanda 软件建立 circRNA-miRNA 相互作用网络。通过 qRT-PCR 测定对照和 TAO 样本中 mRNA 和 circRNA 的表达水平。在 RNA 测序数据预测的所有 16329 个 circRNA 中，有 163 个 circRNA（95 个下调和 68 个上调）在 TAO 样本中差异表达。此外，还鉴定了 607 个差异表达的 mRNA。共表达分析表明，circRNA_14940 与 CCND1 和 TNXB 相关，circRNA_10135 与 PTGFR 相关，circRNA_14936 与 TNFRSF19 相关。上调的 CCND1 参与了 Wnt 信号通路。下调的 TNXB 参与了细胞外基质受体相互作用、焦点黏附以及 PI3K-Akt 信号通路。PTGFR 参与了神经活性配体受体相互作用和钙信号通路。TNFRSF19 参与了细胞因子-细胞因子受体相互作用。在相互作用网络中，circRNA_14936 可以与 hsa-miR-10392-3p 相互作用，circRNA_12367 可以与 hsa-miR-1228-3p 相互作用。此外，通过 qRT-PCR 验证了 MMP2、TNXB、PTGFR、CCND1 和 TNFRSF19 以及 circRNA_14936、circRNA_14940 和 circRNA_12367 的表达变化。总之，差异表达的 circRNA 可能参与了 TAO 的发病机制，我们推测 circRNA_14940-CCND1-Wnt 信号通路可能是一个重要的调控轴。
