Mater Misericordiae University Hospital, Eccles St., Dublin 7, Ireland.
University College Cork, College Road, Cork, Ireland.
Int J Mol Sci. 2021 Nov 15;22(22):12321. doi: 10.3390/ijms222212321.
This study explored the expression of several miRNAs reported to be deregulated in age-related macular degeneration (AMD). Total RNA was isolated from sera from patients with dry AMD ( = 12), wet AMD ( = 14), and controls ( = 10). Forty-two previously investigated miRNAs were selected based on published data and their role in AMD pathogenesis, such as angiogenic and inflammatory effects, and were co-analysed using a miRCURY LNA miRNA SYBR Green PCR kit via quantitative real-time polymerase chain reaction (qRT-PCR) to validate their presence. Unsupervised hierarchical clustering indicated that AMD serum specimens have a different miRNA profile to healthy controls. We successfully validated the differentially regulated miRNAs in serum from AMD patients versus controls. Eight miRNAs (hsa-let-7a-5p, hsa-let-7d-5p, hsa-miR-23a-3p, hsa-miR-301a-3p, hsa-miR-361-5p, hsa-miR-27b-3p, hsa-miR-874-3p, hsa-miR-19b-1-5p) showed higher expression in the serum of dry AMD patients than wet AMD patients and compared with healthy controls. Increased quantities of certain miRNAs in the serum of AMD patients indicate that these miRNAs could potentially serve as diagnostic AMD biomarkers and might be used as future AMD treatment targets. The discovery of significant serum miRNA biomarkers in AMD patients would provide an easy screening tool for at-risk populations.
本研究探讨了几种据报道在年龄相关性黄斑变性（AMD）中失调的 miRNAs 的表达。从干性 AMD 患者（=12）、湿性 AMD 患者（=14）和对照者（=10）的血清中分离总 RNA。基于已发表的数据和它们在 AMD 发病机制中的作用，如血管生成和炎症效应，选择了 42 种先前研究过的 miRNAs，并使用 miRCURY LNA miRNA SYBR Green PCR 试剂盒通过定量实时聚合酶链反应（qRT-PCR）共同分析，以验证它们的存在。无监督层次聚类表明 AMD 血清标本与健康对照者具有不同的 miRNA 谱。我们成功验证了 AMD 患者与对照者血清中差异调节的 miRNAs。八种 miRNAs（hsa-let-7a-5p、hsa-let-7d-5p、hsa-miR-23a-3p、hsa-miR-301a-3p、hsa-miR-361-5p、hsa-miR-27b-3p、hsa-miR-874-3p、hsa-miR-19b-1-5p）在干性 AMD 患者血清中的表达高于湿性 AMD 患者和健康对照者。AMD 患者血清中某些 miRNAs 的含量增加表明，这些 miRNAs 可能作为 AMD 的诊断生物标志物，并可能作为未来 AMD 的治疗靶点。在 AMD 患者中发现有意义的血清 miRNA 生物标志物将为高危人群提供一种简单的筛查工具。
