Department of Ophthalmology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.
Genomic Data Analysis Unit, Hebrew University, Jerusalem, Israel.
Sci Rep. 2016 Jul 4;6:29046. doi: 10.1038/srep29046.
Mononuclear phagocytes (MPs), including monocytes/macrophages, play complex roles in age-related macular degeneration (AMD) pathogenesis. We reported altered gene-expression signature in peripheral blood mononuclear cells from AMD patients, and a chemokine receptor signature on AMD monocytes. To obtain comprehensive understanding of MP involvement, particularly in peripheral circulation in AMD, we performed global gene expression analysis in monocytes. We separated monocytes from treatment-naïve neovascular AMD (nvAMD) patients (n = 14) and age-matched controls (n = 15), and performed microarray and bioinformatics analysis. Quantitative real-time PCR was performed on other sets of nvAMD (n = 25), atrophic AMD (n = 21), and controls (n = 28) for validation. This validated microarray genes (like TMEM176A/B and FOSB) tested, including differences between nvAMD and atrophic AMD. We identified 2,165 differentially-expressed genes (P < 0.05), including 79 genes with log2 fold change ≥1.5 between nvAMD and controls. Functional annotation using DAVID and TANGO demonstrated immune response alterations in AMD monocytes (FDR-P <0.05), validated by randomized data comparison (P < 0.0001). GSEA, ISMARA, and MEME analysis found immune enrichment and specific involved microRNAs. Enrichment of differentially-expressed genes in monocytes was found in retina via SAGE data-mining. These genes were enriched in non-classical vs. classical monocyte subsets (P < 0.05). Therefore, global gene expression analysis in AMD monocytes reveals an altered immune-related signature, further implicating systemic MP activation in AMD.
单核吞噬细胞（MPs），包括单核细胞/巨噬细胞，在年龄相关性黄斑变性（AMD）发病机制中发挥着复杂的作用。我们报道了 AMD 患者外周血单核细胞中改变的基因表达特征，以及 AMD 单核细胞上的趋化因子受体特征。为了全面了解 MP 的参与情况，特别是在 AMD 的外周循环中，我们对单核细胞进行了全基因组基因表达分析。我们从未经治疗的新生血管性 AMD（nvAMD）患者（n=14）和年龄匹配的对照组（n=15）中分离单核细胞，并进行了微阵列和生物信息学分析。在其他 nvAMD（n=25）、萎缩性 AMD（n=21）和对照组（n=28）的患者中进行了定量实时 PCR 验证。该验证的微阵列基因（如 TMEM176A/B 和 FOSB）测试，包括 nvAMD 和萎缩性 AMD 之间的差异。我们确定了 2165 个差异表达基因（P<0.05），其中包括 nvAMD 和对照组之间对数 2 倍变化≥1.5 的 79 个基因。使用 DAVID 和 TANGO 进行的功能注释表明 AMD 单核细胞的免疫反应发生改变（FDR-P<0.05），通过随机数据比较验证（P<0.0001）。GSEA、ISMARA 和 MEME 分析发现免疫富集和特定涉及的 microRNA。通过 SAGE 数据挖掘在外周血单核细胞中发现差异表达基因的富集。这些基因在外周血单核细胞中的富集与非经典单核细胞亚群与经典单核细胞亚群相比（P<0.05）。因此，AMD 单核细胞的全基因组基因表达分析揭示了一个改变的免疫相关特征，进一步表明系统 MPs 的激活与 AMD 有关。
