State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, Guangdong Province, China.
Department of Ophthalmology, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong Province, China.
Invest Ophthalmol Vis Sci. 2024 Feb 1;65(2):25. doi: 10.1167/iovs.65.2.25.
To evaluate the expression of sry-box transcription factor 9 (SOX9) in orbital fibroblasts (OFs) of thyroid eye disease (TED) and to find its potential role and underlying mechanism in orbital fibrosis.
OFs were cultured from orbital connective tissues obtained from patients with TED (n = 10) and healthy controls (n = 6). SOX9 was depleted by small interfering RNA or overexpressed through lentivirus transduction in OFs. Fibroblast contractile activity was measured by collagen gel contraction assay and proliferation was examined by EdU assay. Transcriptomic changes were assessed by RNA sequencing.
The mRNA and protein levels of SOX9 were significantly higher in OFs cultured from patients with TED than those from healthy controls. Extracellular matrix-related genes were down-regulated by SOX9 knockdown and up-regulated by SOX9 overexpression in TED-OFs. SOX9 knockdown significantly decrease the contraction and the antiapoptotic ability of OFs, whereas the overexpression of SOX9 increased the ability of transformation, migration, and proliferation of OFs. SOX9 knockdown suppressed the expression of phosphorylated ERK1/2, whereas its overexpression showed the opposite effect. Epidermal growth factor receptor (EGFR) is one of the notably down-regulated genes screened out by RNA sequencing. Chromatin immunoprecipitation-qPCR demonstrated SOX9 binding to the EGFR promoter.
A high expression of SOX9 was found in TED-OFs. SOX9 can activate OFs via MAPK/ERK1/2 signaling pathway, which in turn promotes proliferation and differentiation of OFs. EGFR was a downstream target gene of SOX9. SOX9/EGFR can be considered as therapeutic targets for the treatment of orbital fibrosis in TED.
评估甲状腺眼病（TED）眼眶成纤维细胞（OFs）中性别决定区 Y 框转录因子 9（SOX9）的表达，寻找其在眼眶纤维化中的潜在作用和潜在机制。
从 TED 患者（n=10）和健康对照者（n=6）眼眶结缔组织中培养 OFs。通过小干扰 RNA 敲低或慢病毒转导过表达 OFs 中的 SOX9。通过胶原凝胶收缩试验测量成纤维细胞收缩活性，通过 EdU 测定法检测增殖。通过 RNA 测序评估转录组变化。
与健康对照组相比，TED 患者来源的 OFs 中 SOX9 的 mRNA 和蛋白水平均显著升高。SOX9 敲低下调了 TED-OFs 中细胞外基质相关基因的表达，而 SOX9 过表达则上调了这些基因的表达。SOX9 敲低显著降低了 OFs 的收缩和抗凋亡能力，而过表达 SOX9 则增加了 OFs 的转化、迁移和增殖能力。SOX9 敲低抑制了磷酸化 ERK1/2 的表达，而过表达则显示出相反的效果。表皮生长因子受体（EGFR）是 RNA 测序筛选出的显著下调基因之一。染色质免疫沉淀-qPCR 表明 SOX9 结合到 EGFR 启动子上。
TED-OFs 中发现 SOX9 高表达。SOX9 可通过 MAPK/ERK1/2 信号通路激活 OFs，进而促进 OFs 的增殖和分化。EGFR 是 SOX9 的下游靶基因。SOX9/EGFR 可作为治疗 TED 眼眶纤维化的治疗靶点。
