Molecular Structure and Functional Genomics Section, National Eye Institute, National Institutes of Health, Bethesda, Maryland, United States of America.
Office of Intramural Research, Center for Information Technology, National Institutes of Health, Bethesda, Maryland, United States of America.
PLoS One. 2024 Sep 26;19(9):e0293383. doi: 10.1371/journal.pone.0293383. eCollection 2024.
ARPE-19 cells are derived from adult human retinal pigment epithelium (RPE). The response of these cells to the stress of serum deprivation mimics some important processes relevant to age-related macular degeneration (AMD). Here we extend the characterization of this response using RNASeq and EGSEA gene set analysis of ARPE-19 cells over nine days of serum deprivation. This experiment confirmed the up-regulation of cholesterol and lipid-associated pathways that increase cholesterol levels in these cells. The gene expression analysis also identified other pathways relevant to AMD progression. There were significant changes in extracellular matrix gene expression, notably a switch from expression of collagen IV, a key component of Bruch's membrane (part of the blood-retina barrier), to expression of a fibrosis-like collagen type I matrix. Changes in the expression profile of the extracellular matrix led to the discovery that amelotin is induced in AMD and is associated with the development of the calcium deposits seen in late-stage geographic atrophy. The transcriptional profiles of other pathways, including inflammation, complement, and coagulation, were also modified, consistent with immune response patterns seen in AMD. As previously noted, the cells resist apoptosis and autophagy but instead initiate a gene expression pattern characteristic of senescence, consistent with the maintenance of barrier function even as other aspects of RPE function are compromised. Other differentially regulated genes were identified that open new avenues for investigation. Our results suggest that ARPE-19 cells maintain significant stress responses characteristic of native RPE that are informative for AMD. As such, they provide a convenient system for discovery and for testing potential therapeutic interventions.
ARPE-19 细胞源自成人视网膜色素上皮 (RPE)。这些细胞对血清剥夺应激的反应模拟了与年龄相关性黄斑变性 (AMD) 相关的一些重要过程。在这里,我们使用 RNA 测序和 EGSEA 基因集分析,对 ARPE-19 细胞在血清剥夺的九天中进行了研究,从而进一步对这种反应进行了表征。该实验证实了胆固醇和脂质相关途径的上调,这些途径会增加细胞中的胆固醇水平。基因表达分析还确定了与 AMD 进展相关的其他途径。细胞外基质基因表达发生了显著变化,特别是从表达布鲁赫膜(血视网膜屏障的一部分)的关键成分 IV 型胶原,转变为表达纤维化样 I 型胶原基质。细胞外基质表达谱的变化导致发现了在 AMD 中诱导的釉蛋白,并与晚期地理萎缩中所见的钙沉积的发展有关。其他途径(包括炎症、补体和凝血)的转录谱也发生了改变,与 AMD 中所见的免疫反应模式一致。如前所述,这些细胞抵抗细胞凋亡和自噬,但会启动衰老的基因表达模式,这与保持屏障功能一致,即使 RPE 功能的其他方面受损。还确定了其他差异调节的基因,为进一步研究开辟了新途径。我们的结果表明,ARPE-19 细胞维持了与天然 RPE 相关的重要应激反应,这些反应对 AMD 具有重要意义。因此,它们为发现和测试潜在的治疗干预措施提供了一个方便的系统。