Shang Peng, Ambrosino Helena, Hoang Johnson, Geng Zhaohui, Zhu Xiaoyu, Shen Shichen, Eminhizer Mark, Hong Elise, Zhang Ming, Qu Jun, Du Jianhai, Montezuma Sandra R, Dutton James R, Ferrington Deborah A
Doheny Eye Institute, Pasadena, CA, 91103, USA.
Stem Cell Institute, University of Minnesota, Minneapolis, MN, 55455, USA.
Free Radic Biol Med. 2024 Nov 20;225:833-845. doi: 10.1016/j.freeradbiomed.2024.10.307. Epub 2024 Nov 2.
Age-related macular degeneration (AMD), the leading cause of central vision loss in the elderly, involves death of the retinal pigment epithelium (RPE) and light-sensing photoreceptors. This multifactorial disease includes contributions from both genetic and environmental risk factors. The current study examined the effect of the Y402H polymorphism of Complement Factor H (CFH, rs1061170) and cigarette smoke, predominant genetic and environmental risk factors associated with AMD. We used targeted and discovery-based approaches to identify genotype-dependent responses to chronic oxidative stress induced by cigarette smoke extract (CSE) in RPE differentiated from induced pluripotent stem cells (iPSC) derived from human donors harboring either the low risk (LR) or high risk (HR) CFH genotype. Chronic CSE altered the metabolic profile in both LR and HR iPSC-RPE and caused a dose-dependent reduction in mitochondrial function despite an increase in mitochondrial content. Notably, cells with the HR CFH SNP showed a greater reduction in maximal respiration and ATP production. Significant genotype-dependent changes in the proteome were observed for HR RPE at baseline (cytoskeleton, MAPK signaling) and after CSE exposure, where a less robust upregulation of the antioxidants and significant downregulation in proteins involved in nucleic acid metabolism and membrane trafficking were noted compared to LR cells. In LR cells, uniquely upregulated proteins were involved in lipid metabolism and chemical detoxification. These genotype-dependent differences at baseline and in response to chronic CSE exposure suggest a broader role for CFH in modulating the response to oxidative stress in RPE and provides insight into the interaction between environmental and genetic factors in AMD pathogenesis.
年龄相关性黄斑变性（AMD）是老年人中心视力丧失的主要原因，涉及视网膜色素上皮（RPE）和感光光感受器的死亡。这种多因素疾病包括遗传和环境风险因素的影响。本研究检测了补体因子H（CFH，rs1061170）的Y402H多态性和香烟烟雾（AMD主要的遗传和环境风险因素）的作用。我们采用靶向和基于发现的方法，以确定在源自携带低风险（LR）或高风险（HR）CFH基因型的人类供体的诱导多能干细胞（iPSC）分化而来的RPE中，对香烟烟雾提取物（CSE）诱导的慢性氧化应激的基因型依赖性反应。慢性CSE改变了LR和HR iPSC-RPE的代谢谱，尽管线粒体含量增加，但导致线粒体功能呈剂量依赖性降低。值得注意的是，具有HR CFH单核苷酸多态性的细胞在最大呼吸和ATP产生方面的降低幅度更大。在基线时（细胞骨架、丝裂原活化蛋白激酶信号传导）以及CSE暴露后，观察到HR RPE的蛋白质组存在显著的基因型依赖性变化，与LR细胞相比，HR RPE中抗氧化剂的上调不太明显，参与核酸代谢和膜运输的蛋白质显著下调。在LR细胞中，独特上调的蛋白质参与脂质代谢和化学解毒。这些基线时以及对慢性CSE暴露反应中的基因型依赖性差异表明，CFH在调节RPE对氧化应激的反应中具有更广泛的作用，并为AMD发病机制中环境和遗传因素之间的相互作用提供了见解。
