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近视患者房水来源外泌体 miRNAs 的表达谱分析。

Expression Profiling of Exosomal miRNAs Derived from the Aqueous Humor of Myopia Patients.

机构信息

Cancer Progression Research Center, National Yang-Ming University.

Department of Ophthalmology, Taipei City Hospital.

出版信息

Tohoku J Exp Med. 2019 Nov;249(3):213-221. doi: 10.1620/tjem.249.213.

DOI:10.1620/tjem.249.213
PMID:31776299
Abstract

Myopia is the most common refractive disorder in Eastern Asia. The development of myopia is associated with the cooperation of various ocular tissues. Exosomes in the aqueous humor (AH) have been implicated to modulate intracellular communications by transferring exosomal miRNAs and proteins between cells. These exosomal miRNAs and proteins are likely involved in the pathogenesis of various eye diseases. In this study, we aimed to explore human exosomal miRNA profiles and their roles in myopia development. AH samples were collected from 16 patients (8 myopia and 8 control) undergoing routine cataract surgeries. Exosomes were isolated from AH of each individual using the ExoQuick solution. The numbers and sizes of exosomes were not significantly different between the myopia and control groups. The individual exosomes of the same group were pooled to purify RNA. Unexpectedly, the myopia group contained 2.78-fold total RNA amount than that in the control group. Thereafter, miRNA profiles were analyzed using the OpenArray system. We thus found 15 myopia-specific miRNAs and four myopia-absent miRNAs. By using bioinformatics analysis, we identified six well-known myopia-associated genes that are potential targets of five myopia-specific miRNAs (has-miR-582-3p, has-miR-17-5p, has-miR-885-3p, has-miR-19b-3p, and has-miR-450b-5p). These genes are cholinergic receptor muscarinic 2 (CHRM2), cyclic nucleotide-gated channel beta 3 (CNGB3), vascular endothelial growth factor A (VEGFA), adenosine A2a receptor (ADORA2A), insulin-like growth factor 1 (IGF1), and lumican (LUM). Moreover, CHRM2 may be a target of myopia-absent miRNA (has-miR-378a-5p). In conclusion, we show the expression profiles of AH-derived exosomal miRNAs and their potential roles in myopia development.

摘要

近视是东亚最常见的屈光不正。近视的发展与各种眼组织的合作有关。房水中的外泌体(AH)已被认为通过在细胞之间转移外泌体 miRNA 和蛋白质来调节细胞内通讯。这些外泌体 miRNA 和蛋白质可能参与各种眼病的发病机制。在这项研究中,我们旨在探索人外泌体 miRNA 谱及其在近视发展中的作用。从 16 名(8 名近视和 8 名对照)接受常规白内障手术的患者中采集 AH 样本。使用 ExoQuick 溶液从每个人的 AH 中分离外泌体。近视组和对照组之间的外泌体数量和大小没有显著差异。同一组的个体外泌体被汇集以纯化 RNA。出乎意料的是,近视组的总 RNA 量比对照组多 2.78 倍。此后,使用 OpenArray 系统分析 miRNA 谱。因此,我们发现了 15 种近视特异性 miRNA 和 4 种近视缺失 miRNA。通过生物信息学分析,我们确定了六个已知的近视相关基因,它们是五个近视特异性 miRNA 的潜在靶点(has-miR-582-3p、has-miR-17-5p、has-miR-885-3p、has-miR-19b-3p 和 has-miR-450b-5p)。这些基因是胆碱能受体毒蕈碱 2(CHRM2)、环核苷酸门控通道β3(CNGB3)、血管内皮生长因子 A(VEGFA)、腺苷 A2a 受体(ADORA2A)、胰岛素样生长因子 1(IGF1)和亮氨酸(LUM)。此外,CHRM2 可能是近视缺失 miRNA(has-miR-378a-5p)的靶点。总之,我们展示了 AH 衍生的外泌体 miRNA 的表达谱及其在近视发展中的潜在作用。

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