Supplementary information files for Retinal pigment epithelium extracellular vesicles are potent inducers of age-related macular degeneration disease phenotype in the outer retina

Supplementary files for article Retinal pigment epithelium extracellular vesicles are potent inducers of age-related macular degeneration disease phenotype in the outer retina <br> Age-related macular degeneration (AMD) is a leading cause of blindness. Vision loss is caused by the retinal pigment epithelium (RPE) and photoreceptors atrophy and/or retinal and choroidal angiogenesis. Here we use AMD patient-specific RPE cells with the Complement Factor H Y402H high-risk polymorphism to perform a comprehensive analysis of extracellular vesicles (EVs), their cargo and role in disease pathology. We show that AMD RPE is characterised by enhanced polarised EV secretion. Multi-omics analyses demonstrate that AMD RPE EVs carry RNA, proteins and lipids, which mediate key AMD features including oxidative stress, cytoskeletal dysfunction, angiogenesis and drusen accumulation. Moreover, AMD RPE EVs induce amyloid fibril formation, revealing their role in drusen formation. We demonstrate that exposure of control RPE to AMD RPE apical EVs leads to the acquisition of AMD features such as stress vacuoles, cytoskeletal destabilization and abnormalities in the morphology of the nucleus. Retinal organoid treatment with apical AMD RPE EVs leads to disrupted neuroepithelium and the appearance of cytoprotective alpha B crystallin immunopositive cells, with some co-expressing retinal progenitor cell markers Pax6/Vsx2, suggesting injury-induced regenerative pathways activation. These findings indicate that AMD RPE EVs are potent inducers of AMD phenotype in the neighbouring RPE and retinal cells.

本数据集为论文《视网膜色素上皮细胞外囊泡是外视网膜年龄相关性黄斑变性表型的强效诱导剂》的补充材料。
年龄相关性黄斑变性（age-related macular degeneration, AMD）是全球首位致盲性眼病，其视力丧失由视网膜色素上皮（Retinal pigment epithelium, RPE）与光感受器萎缩，以及视网膜、脉络膜血管新生所引发。本研究使用携带补体因子H Y402H高风险多态性的AMD患者特异性RPE细胞，对细胞外囊泡（extracellular vesicles, EVs）、其携带的内容物及其在疾病病理中的作用开展全面分析。
研究发现，AMD来源的RPE细胞以极化分泌EVs增强为典型特征。多组学分析表明，AMD RPE来源的EVs携带有RNA、蛋白质与脂质，这些物质可介导AMD的关键病理特征，包括氧化应激、细胞骨架功能异常、血管生成及玻璃膜疣沉积。此外，AMD RPE来源的EVs可诱导淀粉样纤维形成，揭示了其在玻璃膜疣形成中的作用。
研究证实，将正常RPE细胞暴露于AMD RPE的顶侧EVs后，正常细胞会获得AMD相关表型，如应激空泡形成、细胞骨架失稳及细胞核形态异常。使用顶侧AMD RPE来源的EVs处理视网膜类器官（retinal organoid），可导致神经上皮结构紊乱，并出现具有细胞保护作用的αB晶状体蛋白（alpha B crystallin）免疫阳性细胞，其中部分细胞共表达视网膜祖细胞标志物Pax6与Vsx2，提示损伤诱导的再生通路被激活。
上述研究结果表明，AMD RPE来源的EVs是邻近RPE细胞与视网膜细胞AMD表型的强效诱导因子。