microRNA Profiling in Glaucoma Eyes With Varying Degrees of Optic Neuropathy by Using Next-Generation Sequencing
microRNA Profiling in Glaucoma Eyes With Varying Degrees of Optic Neuropathy by Using Next-Generation Sequencing
复制标题
使用下一代测序对患有不同程度视神经病变的青光眼眼进行 microRNA 分析。
DOI：
10.1167/iovs.17-23599
复制
发表时间：
2018-06-01
影响因子：
4.4
通讯作者：
Zhang, Xiulan
中科院分区：
文献类型：
--
作者：
Liu, Yaoming;Chen, Yang;Zhang, Xiulan
目的.利用新一代测序技术探讨microRNA（miRNA）的表达谱及其在青光眼视神经病变中的作用。术前采集19只原发性开角型青光眼（POAG）眼和17只白内障眼的房水（AH）样本。对从18个AH样本中提取的RNA样本进行下一代测序，并对具有足够干净数据输出的样本应用生物信息学方法。另外18份样本用于定量PCR验证测序结果。总共有12个（6个POAG和6个白内障对照）样本在测序后具有足够的干净数据输出，用于进一步的数据分析。在POAG和白内障对照组中分别检测到466和480个成熟miRNAs。其中，164个miRNAs在所有POAG样本中检测到，96个miRNAs在所有白内障对照样本中检测到。此外，88个miRNAs被鉴定为在POAG和白内障对照眼之间差异表达。此外，有16种miRNAs在重度视野损害的POAG眼和中度视野损害的POAG眼中表达不同。预测这种差异表达调节硫胺素代谢、嘌呤代谢和转录失调。hsa-miR-184、hsa-miR-486 - 5p和hsa-miR-93 - 5p的相对表达模式通过定量PCR证实。本研究全面展示了POAG眼AH中miRNA的表达谱，尤其是不同程度视野损害眼中miRNA的差异表达，与潜在的miRNA相关通路一起，为POAG的发病和进展提供了新的靶点。
PURPOSE. To explore the microRNA (miRNA) profile and its putative role in glaucomatous optic neuropathy by using next-generation sequencing.METHODS. Aqueous humor (AH) samples were collected from 19 primary open-angle glaucoma (POAG) eyes and 17 cataract eyes before surgery. Next-generation sequencing was performed for RNA samples extracted from 18 AH samples, and the bioinformatics approach was applied for samples with adequate clean data output. The other 18 samples were used for quantitative PCR validation of sequencing results.RESULTS. In total, 12 (six POAG and six cataract controls) samples with sufficient clean data output after sequencing were used for further data analysis. Four hundred sixty-six and 480 mature miRNAs were detected in the POAG and cataract control groups, respectively. Among them, 164 miRNAs were detected in all POAG samples, and 96 miRNAs were detected in all cataract control samples. Furthermore, 88 miRNAs were identified as differently expressed between POAG and cataract control eyes. In addition, 16 miRNAs were differently expressed between POAG eyes with severe visual field damage and eyes with moderate visual field damage. This differential expression was predicted to regulate thiamine metabolism, purine metabolism, and transcriptional misregulation. Relative expression patterns of hsa-miR-184, hsa-miR-486-5p, and hsa-miR-93-5p were confirmed by quantitative PCR.CONCLUSIONS. This study comprehensively demonstrated the miRNA expression profile in the AH of POAG eyes, especially the differential expression of miRNA in eyes with varying degrees of visual field damage, which, together with the underlying miRNA-related pathways, indicate new targets for the pathogenesis and progression of POAG.