The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Lab of Ophthalmology, Chongqing Eye Institute, Chongqing Branch of National Clinical Research Center for Ocular Diseases, Chongqing, P. R. China.
Curr Eye Res. 2022 Aug;47(8):1156-1164. doi: 10.1080/02713683.2022.2071945. Epub 2022 May 16.
The differential gene expression of neural precursor cell expressed developmentally downregulated 9 (NEDD9) in human trabecular meshwork (HTM) cells after dexamethasone (Dex) treatment was confirmed through gene expression profiling. However, the regulatory mechanism of NEDD9 expression in HTM cells remains unknown. In this study, we investigated NEDD9 expression in HTM cells and gained a better understanding of glucocorticoid-induced glaucoma (GIG) pathophysiology.
The Gene Expression Omnibus database and GEO2R tool were used to identify differentially expressed genes in the GSE37474 and GSE124114 datasets, and NEDD9 gene expression was found to be upregulated. Human corneoscleral segments and HTM cells were treated with 100 nM Dex or an equal volume of ethanol (0.01%) for 7 days. NEDD9 expression in TM tissues was evaluated by immunohistochemistry, and NEED9 expression in HTM cells was confirmed by RT-qPCR and western blotting. HTM cell adhesive behaviors were assessed with a cell adhesion detection kit. NEDD9 expression was knocked down with short hairpin RNA in HTM cells, and FAK/Src signaling pathway activation was found to be regulated by NEDD9.
After 7 days of HTM cell Dex treatment, NEDD9 expression was upregulated to be approximately twice that of control. FAK, Src, phospho-FAK, and phospho-Src expression in Dex-treated HTM cells was markedly increased. Downregulation of NEDD9 expression reduced HTM cell adhesion to the surface of culture wells and simultaneously led to a reduction in FAK, Src, phospho-FAK and phospho-Src expression.
NEDD9 expression is upregulated in HTM cells after Dex treatment and promotes HTM cell adhesion. These findings underscore the contribution of NEDD9 overexpression to altered HTM cell adhesion during glucocorticoid therapy and may play a key role in GIG pathological progression. Considering the similarity between GIG and primary open-angle glaucoma (POAG), our findings suggest that targeting NEDD9 may be a new therapeutic strategy for POAG patients.
通过基因表达谱分析，证实了在人眼小梁网（HTM）细胞中，地塞米松（Dex）处理后神经前体细胞表达的发育下调 9（NEDD9）的差异基因表达。然而，NEDD9 在 HTM 细胞中的表达调控机制尚不清楚。本研究旨在探讨 NEDD9 在 HTM 细胞中的表达情况，以期更好地了解糖皮质激素诱导性青光眼（GIG）的病理生理学。
使用基因表达综合数据库和 GEO2R 工具，在 GSE37474 和 GSE124114 数据集，发现 NEDD9 基因表达上调。用 100nM Dex 或等体积乙醇（0.01%）处理人角膜巩膜段和 HTM 细胞 7 天。通过免疫组织化学评估 TM 组织中的 NEDD9 表达，通过 RT-qPCR 和 Western blot 确认 HTM 细胞中的 NEDD9 表达。用细胞黏附检测试剂盒评估 HTM 细胞的黏附行为。用短发夹 RNA 敲低 HTM 细胞中的 NEDD9 表达，发现 NEDD9 调节 FAK/Src 信号通路的激活。
Dex 处理 HTM 细胞 7 天后，NEDD9 表达上调约 2 倍。Dex 处理的 HTM 细胞中 FAK、Src、磷酸化 FAK 和磷酸化 Src 的表达明显增加。下调 NEDD9 表达可降低 HTM 细胞黏附至培养孔表面，同时降低 FAK、Src、磷酸化 FAK 和磷酸化 Src 的表达。
Dex 处理后 HTM 细胞中 NEDD9 表达上调，并促进 HTM 细胞黏附。这些发现强调了 NEDD9 过表达在糖皮质激素治疗期间改变 HTM 细胞黏附的作用，可能在 GIG 病理进展中起关键作用。考虑到 GIG 和原发性开角型青光眼（POAG）之间的相似性，我们的研究结果表明，靶向 NEDD9 可能是 POAG 患者的一种新的治疗策略。
