Circulating exosomes from Alzheimer's disease suppress VE-cadherin expression and induce barrier dysfunction in recipient brain microvascular endothelial cell.
Circulating exosomes from Alzheimer's disease suppress VE-cadherin expression and induce barrier dysfunction in recipient brain microvascular endothelial cell.
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来自阿尔茨海默病的循环外泌体抑制 VE-钙粘蛋白表达并诱导受体脑微血管内皮细胞屏障功能障碍。
DOI：
10.1101/2023.04.03.535441
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Gong,Bin
中科院分区：
文献类型：
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作者：
Bei,Jiani;Miranda-Morales,ErnestoG;Gan,Qini;Qiu,Yuan;Husseinzadeh,Sorosh;Liew,JiaYi;Chang,Qing;Krishnan,Balaji;Gaitas,Angelo;Yuan,Subo;Felicella,Michelle;Qiu,WeiQiao;Fang,Xiang;Gong,Bin
血脑屏障（BBB）破坏是阿尔茨海默病（AD）进展和病理的一个组成部分。血脑屏障功能障碍的主要原因是脑微血管内皮细胞（BMEC）紧密连接蛋白减少或紊乱或粘附。尽管越来越多的证据表明，AD患者的bmec中紧密连接的破坏，粘附连接在AD患者血脑屏障功能障碍中的功能作用仍然未知。衰老细胞分泌的外泌体具有独特的特征，有助于调节受体细胞的表型。然而，尚不清楚这些外泌体是否以及如何导致AD患者BMEC功能障碍。目的探讨AD循环外泌体及其RNA载体在AD脑内皮功能障碍中的潜在作用。方法采用尺寸排除色谱技术从5例AD患者血清中分离外泌体，并与年龄和性别匹配的认知正常对照进行比较。我们用纳米颗粒跟踪分析和原子力显微镜验证了分离的外泌体的质量和粒径。我们测量了bmec内皮屏障的生物力学性质，活体bmec之间的横向结合力，采用流体力错误复制。我们在BMEC培养物和采用原代人BMEC和周细胞的3D血脑屏障模型中可视化了关键粘附连接蛋白VE-cadherin的细胞旁表达，并使用共聚焦显微镜对其进行了评估。我们还检测了5例AD患者和5例年龄和性别匹配的认知正常对照组脑组织中的ve -钙粘蛋白信号。结果我们发现AD患者循环外泌体抑制VE-cadherin的细胞旁表达水平，损害受体bmec的屏障功能。免疫染色分析显示，AD循环外泌体在微血管小管形成的3D模型中破坏ve -钙粘蛋白的完整性。我们发现AD的循环外泌体根据RNA货物削弱血脑屏障。同时，我们观察到，与正常对照相比，AD大脑微血管ve -钙粘蛋白表达减少。结论本研究利用vitroandex vivomo模型，说明AD患者循环外泌体以外泌体rna依赖的方式介导血脑屏障受体BMEC粘附连接的损伤作用。这提示了外周衰老外泌体与AD风险的新机制。
BackgroundBlood-brain barrier (BBB) breakdown is a component of the progression and pathology of Alzheimer’s disease (AD). BBB dysfunction is primarily caused by reduced or disorganized tight junction or adherens junction proteins of brain microvascular endothelial cell (BMEC). While there is growing evidence of tight junction disruption in BMECs in AD, the functional role of adherens junctions during BBB dysfunction in AD remains unknown. Exosomes secreted from senescent cells have unique characteristics and contribute to modulating the phenotype of recipient cells. However, it remains unknown if and how these exosomes cause BMEC dysfunction in AD.ObjectivesThis study aimed to investigate the potential roles of AD circulating exosomes and their RNA cargos in brain endothelial dysfunction in AD.MethodsWe isolated exosomes from sera of five cases of AD compared with age- and sex-matched cognitively normal controls using size-exclusion chromatography technology. We validated the qualities and particle sizes of isolated exosomes with nanoparticle tracking analysis and atomic force microscopy. We measured the biomechanical natures of the endothelial barrier of BMECs, the lateral binding forces between live BMECs, using fluidic force miscopy. We visualized the paracellular expressions of the key adherens junction protein VE-cadherin in BMEC cultures and a 3D BBB model that employs primary human BMECs and pericytes with immunostaining and evaluated them using confocal microscopy. We also examined the VE-cadherin signal in brain tissues from five cases of AD and five age- and sex-matched cognitively normal controls.ResultsWe found that circulating exosomes from AD patients suppress the paracellular expression levels of VE-cadherin and impair the barrier function of recipient BMECs. Immunostaining analysis showed that AD circulating exosomes damage VE-cadherin integrity in a 3D model of microvascular tubule formation. We found that circulating exosomes in AD weaken the BBB depending on the RNA cargos. In parallel, we observed that microvascular VE-cadherin expression is diminished in AD brains compared to normal controls.ConclusionUsingin vitroandex vivomodels, our study illustrates that circulating exosomes from AD patients play a significant role in mediating the damage effect on adherens junction of recipient BMEC of the BBB in an exosomal RNA-dependent manner. This suggests a novel mechanism of peripheral senescent exosomes for AD risk.