Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration.
Altered bioenergetics and enhanced resistance to oxidative stress in human retinal pigment epithelial cells from donors with age-related macular degeneration.
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DOI：
10.1016/j.redox.2017.05.015
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发表时间：
2017-10
期刊：
影响因子：
11.4
通讯作者：
Montezuma SR
中科院分区：
文献类型：
--
作者：
Ferrington DA;Ebeling MC;Kapphahn RJ;Terluk MR;Fisher CR;Polanco JR;Roehrich H;Leary MM;Geng Z;Dutton JR;Montezuma SR
视网膜相关性黄斑变性（AMD）是老年人失明的主要原因。已经提出视网膜色素上皮（RPE）中的线粒体缺陷是AMD病理学的基础。为了验证这一想法，我们开发了RPE的原代培养物，以询问来自AMD供体的RPE与健康年龄匹配的供体相比是否在代谢谱上有所不同。对基因表达、蛋白质含量和RPE功能的分析表明，这些培养的细胞复制了体内RPE的许多基本特征。使用Seahorse细胞外通量分析仪测量生物能量学，我们观察到与健康供体相比，来自AMD供体的RPE表现出降低的线粒体和糖酵解功能。与健康供体的细胞相比，AMD供体的RPE对这两种能量产生途径的氧化失活也更具抵抗力，并且对氧化诱导的细胞死亡不太敏感。对生物能量学和抗氧化应激性差异的潜在机制的研究表明，来自AMD供体的RPE在响应氧化挑战时具有增加的PGC1α蛋白以及多个基因的差异表达。基于我们的数据，我们提出，培养的RPE从捐助者表型的存在或不存在的AMD提供了一个很好的模型系统，用于研究“AMD在一个盘子”。我们的结果与以下想法一致：（i）RPE中的生物能量学危机有助于AMD病理学，以及（ii）体内的患病环境导致保留在体外的细胞概况的变化。来自成年供体的视网膜色素上皮（RPE）的原代培养物在体内复制了RPE的许多特征。AMD供体RPE中线粒体和糖酵解功能的降低表明生物能量危机有助于AMD病理学。体内的患病环境引起了保留在体外的细胞谱的变化。AMD供体RPE中改变的生物能量学和氧化表明这些培养物是研究“培养皿中的AMD”的良好模型。
Age-related macular degeneration (AMD) is the leading cause of blindness among older adults. It has been suggested that mitochondrial defects in the retinal pigment epithelium (RPE) underlies AMD pathology. To test this idea, we developed primary cultures of RPE to ask whether RPE from donors with AMD differ in their metabolic profile compared with healthy age-matched donors. Analysis of gene expression, protein content, and RPE function showed that these cultured cells replicated many of the cardinal features of RPE in vivo. Using the Seahorse Extracellular Flux Analyzer to measure bioenergetics, we observed RPE from donors with AMD exhibited reduced mitochondrial and glycolytic function compared with healthy donors. RPE from AMD donors were also more resistant to oxidative inactivation of these two energy-producing pathways and were less susceptible to oxidation-induced cell death compared with cells from healthy donors. Investigation of the potential mechanism responsible for differences in bioenergetics and resistance to oxidative stress showed RPE from AMD donors had increased PGC1α protein as well as differential expression of multiple genes in response to an oxidative challenge. Based on our data, we propose that cultured RPE from donors phenotyped for the presence or absence of AMD provides an excellent model system for studying “AMD in a dish”. Our results are consistent with the ideas that (i) a bioenergetics crisis in the RPE contributes to AMD pathology, and (ii) the diseased environment in vivo causes changes in the cellular profile that are retained in vitro. Primary cultures of retinal pigment epithelium (RPE) from adult donors replicate many features of RPE in vivo. Decreased mitochondrial and glycolytic function in AMD donor RPE suggests a bioenergetic crisis contributes to AMD pathology. The diseased environment in vivo caused changes in the cellular profile that are retained in vitro. Altered bioenergetics and oxidation in AMD donor RPE suggest these cultures are a good model for studying "AMD in a dish".