Modeling age‐related macular degeneration (AMD) is challenging, because it is a multifactorial disease. To focus on interactions between the retinal pigment epithelium (RPE) and Bruch's membrane, we generated RPE from AMD patients and used an altered extracellular matrix (ECM) that models aged Bruch's membrane. Induced pluripotent stem cells (iPSCs) were generated from fibroblasts isolated from AMD patients or age‐matched (normal) controls. RPE derived from iPSCs were analyzed by morphology, marker expression, transepithelial electrical resistance (TER), and phagocytosis of rod photoreceptor outer segments. Cell attachment and viability was tested on nitrite‐modified ECM, a typical modification of aged Bruch's membrane. DNA microarrays with hierarchical clustering and analysis of mitochondrial function were used to elucidate possible mechanisms for the observed phenotypes. Differentiated RPE displayed cell‐specific morphology and markers. The TER and phagocytic capacity were similar among iPSC‐derived RPE cultures. However, distinct clusters were found for the transcriptomes of AMD and control iPSC‐derived RPE. AMD‐derived iPSC‐RPE downregulated genes responsible for metabolic‐related pathways and cell attachment. AMD‐derived iPSC‐RPE exhibited reduced mitochondrial respiration and ability to attach and survive on nitrite‐modified ECM. Cells that did attach induced the expression of complement genes. Despite reprogramming, iPSC derived from AMD patients yielded RPE with a transcriptome that is distinct from that of age‐matched controls. When challenged with an AMD‐like modification of Bruch's membrane, AMD‐derived iPSC‐RPE activated the complement immune system.

中文翻译：

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来自年龄相关性黄斑变性患者的干细胞衍生的视网膜色素上皮表现出代谢和基质相互作用降低。

模拟年龄相关性黄斑变性 (AMD) 具有挑战性，因为它是一种多因素疾病。为了关注视网膜色素上皮 (RPE) 和 Bruch 膜之间的相互作用，我们从 AMD 患者中生成了 RPE，并使用了一种改变的细胞外基质 (ECM) 来模拟老化的 Bruch 膜。诱导多能干细胞 (iPSC) 是从 AMD 患者或年龄匹配（正常）对照的成纤维细胞中产生的。通过形态学、标记表达、跨上皮电阻 (TER) 和杆状光感受器外段的吞噬作用分析了来自 iPSC 的 RPE。细胞附着和活力在亚硝酸盐修饰的 ECM 上进行测试，这是对老化的布鲁赫膜的典型修饰。具有层次聚类和线粒体功能分析的 DNA 微阵列用于阐明观察到的表型的可能机制。分化的 RPE 显示细胞特异性形态和标记。iPSC 衍生的 RPE 培养物中的 TER 和吞噬能力相似。然而，在 AMD 和对照 iPSC 衍生的 RPE 的转录组中发现了不同的簇。AMD 衍生的 iPSC-RPE 下调负责代谢相关途径和细胞附着的基因。AMD 衍生的 iPSC-RPE 表现出线粒体呼吸减少以及在亚硝酸盐修饰的 ECM 上附着和存活的能力。确实附着的细胞诱导了补体基因的表达。尽管进行了重编程，但源自 AMD 患者的 iPSC 产生的 RPE 具有与年龄匹配的对照不同的转录组。