通过房水长链非编码 RNA 谱分析鉴定原发性开角型青光眼的潜在生物标志物。

Potential Biomarkers for Primary Open-Angle Glaucoma Identified by Long Noncoding RNA Profiling in the Aqueous Humor.

机构信息

Department of Ophthalmology, The Second Xiangya Hospital, Central South University, Changsha, People's Republic of China; Hunan Clinical Research Center of Ophthalmic Disease, Changsha, People's Republic of China.

Department of Ophthalmology, Institute for Human Genetics, University of California, San Francisco School of Medicine, San Francisco, California; Department of Anatomy, Institute for Human Genetics, University of California, San Francisco School of Medicine, San Francisco, California.

出版信息

Am J Pathol. 2019 Apr;189(4):739-752. doi: 10.1016/j.ajpath.2018.12.011. Epub 2019 Jan 22.

DOI:10.1016/j.ajpath.2018.12.011

PMID:30677397

Abstract

This study aimed to identify potential biomarkers for primary open-angle glaucoma (POAG) diagnosis. First, long noncoding RNA (lncRNA) and mRNA expression profiles in the aqueous humor (AH) from 10 POAG and 10 control patients were accessed by microarray analyses. Coding-noncoding gene coexpression networks were drawn to predict potential lncRNA functions. lncRNA T267384, ENST00000607393, and T342877 expression levels were further tested by real-time quantitative PCR in AH from 29 POAG and 30 cataract patients, in iris tissues from 16 POAG patients and 10 controls, and in plasma from 49 POAG patients and 55 healthy controls. Finally, ENST00000607393 function was characterized in an in vitro model of cell calcification. A total of 3627 lncRNAs and 2228 mRNAs in the AH of POAG patients were significantly up-regulated, and 1520 lncRNAs and 820 mRNAs were significantly down-regulated. Seven lncRNAs showed positive correlation with glaucoma-associated gene, bone morphogenetic protein 2. Moreover, real-time quantitative RT-PCR confirmed that T267384, ENST00000607393, and T342877 expression levels were significantly higher in the AH from a different cohort of POAG patients. ENST00000607393 was also significantly higher in the iris and plasma of POAG patients. Last, ENST00000607393 knockdown alleviated calcification of primary human trabecular meshwork cells in vitro. Therefore, lncRNAs T267384, ENST00000607393, and T342877 may be potential biomarkers for POAG diagnosis. ENST00000607393 might be a new therapeutic target for trabecular meshwork calcification.

摘要

本研究旨在鉴定原发性开角型青光眼（POAG）诊断的潜在生物标志物。首先，通过微阵列分析获取 10 例 POAG 患者和 10 例对照患者房水中的长链非编码 RNA（lncRNA）和 mRNA 表达谱。绘制编码-非编码基因共表达网络以预测潜在 lncRNA 功能。进一步通过实时定量 PCR 检测房水中 lncRNA T267384、ENST00000607393 和 T342877 的表达水平，检测对象为 29 例 POAG 患者和 30 例白内障患者、16 例 POAG 患者和 10 例对照患者的虹膜组织以及 49 例 POAG 患者和 55 例健康对照者的血浆。最后，在细胞钙化的体外模型中对 ENST00000607393 功能进行了表征。POAG 患者房水中的 3627 个 lncRNA 和 2228 个 mRNA 显著上调，1520 个 lncRNA 和 820 个 mRNA 显著下调。7 个 lncRNA 与骨形态发生蛋白 2 相关的青光眼基因呈正相关。此外，实时定量 RT-PCR 证实不同队列的 POAG 患者房水中 T267384、ENST00000607393 和 T342877 的表达水平明显更高。POAG 患者的虹膜和血浆中 ENST00000607393 的水平也明显更高。最后，ENST00000607393 敲低可减轻体外原代人小梁网细胞的钙化。因此，lncRNA T267384、ENST00000607393 和 T342877 可能是 POAG 诊断的潜在生物标志物。ENST00000607393 可能是小梁网钙化的新治疗靶点。