年龄相关性黄斑变性（AMD）是致盲的主要原因。视力损失是由视网膜色素上皮（RPE）和光感受器的萎缩和/或视网膜和脉络膜新生血管引起的。在这里，我们使用AMD患者特异性的RPE细胞，具有补体因子H Y402H高风险多态性，对细胞外囊泡（EVs）及其载荷和疾病病理学的综合分析进行了研究。我们发现AMD RPE以增强的极化EV分泌为特征。多组学分析表明AMD RPE EV携带RNA、蛋白质和脂质，介导关键的AMD特征，包括氧化应激、细胞骨架功能障碍、血管生成和Drusen沉积。此外，AMD RPE EVs诱导淀粉样纤维形成，揭示了它们在Drusen形成中的作用。我们证明，暴露于AMD RPE顶部EV的控制RPE会导致获得AMD特征，例如压力空泡、细胞骨架不稳定和细胞核形态的异常。视网膜类器官处理顶部AMD RPE EV会导致神经上皮的破坏和出现细胞保护的αB结晶蛋白免疫阳性细胞，其中一些同时表达视网膜祖细胞标记物Pax6/Vsx2，表明损伤诱导再生途径的激活。这些发现表明AMD RPE EV是邻近RPE和视网膜细胞的强效诱导器AMD表型的原因。©2022作者。由Wiley Periodicals，LLC代表国际细胞外囊泡协会出版的《细胞外囊泡杂志》。

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.© 2022 The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular Vesicles.