神经元衍生细胞外小泡中的突触蛋白作为阿尔茨海默病的生物标志物:新方法和概念的临床证明。

Erez Eitan, Tricia Thornton-Wells, Katya Elgart, Eren Erden, Eve Gershun, Amir Levine, Olga Volpert, Mitra Azadeh, Daniel G Smith, Dimitrios Kapogiannis
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引用次数: 0

摘要

目的:血液生物标志物可以改善阿尔茨海默病(AD)的药物开发及其治疗。血浆中神经元衍生的细胞外小泡(NDEV)为开发新的生物标志物提供了一个微创平台,可用于监测AD涉及的各种致病过程。然而,NDEV仅占循环细胞外小囊泡(EV)的一小部分。大多数已发表的研究都利用L1细胞粘附分子(L1CAM)进行NDEV免疫捕获。我们旨在开发和优化一种替代的、高度特异性的免疫亲和方法,以富集血液NDEV,用于生物标志物的开发。方法:在筛选多种神经元抗原后,我们使用抗生长相关蛋白(GAP)43和神经胶质蛋白3(NLGN3)的抗体实现了高亲和力和特异性的NDEV捕获。通过电子显微镜、蛋白质印迹和蛋白质组学证实了捕获材料的EV身份。神经元来源的特异性通过显示神经元标记物(蛋白质、mRNA)的富集和掺入的神经元EVs的回收来证明。我们从两组早期AD患者(N=19和N=40)和对照组(N=20和N=19)的血浆样本中回顾性分离NDEV,并测量p181 Tau、淀粉样蛋白β(Aβ)42、脑源性神经营养因子(BDNF)、前体脑源性营养因子(proBDNF),结果:AD样本中p181 Tau、Aβ42和NRGN升高,而proBDNF、GluR2、PSD95、GAP43和syntaxin-1降低。p181 Tau、前BDNF和GluR2的差异在多次比较校正中幸存下来,并与认知评分相关。一个包含生物标志物的模型正确地对94.7%的AD参与者和61.5%的对照参与者进行了分类。观察到的NDEV相关生物标志物的差异与先前的研究结果一致。结论:GAP43和NLGN3免疫捕获分离NDEV为AD生物标志物的开发提供了一个强大的新平台,适合大规模验证。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Synaptic proteins in neuron-derived extracellular vesicles as biomarkers for Alzheimer's disease: novel methodology and clinical proof of concept.

Aims: Blood biomarkers can improve drug development for Alzheimer's disease (AD) and its treatment. Neuron-derived extracellular vesicles (NDEVs) in plasma offer a minimally invasive platform for developing novel biomarkers that may be used to monitor the diverse pathogenic processes involved in AD. However, NDEVs comprise only a minor fraction of circulating extracellular vesicles (EVs). Most published studies have leveraged the L1 cell adhesion molecule (L1CAM) for NDEV immunocapture. We aimed to develop and optimize an alternative, highly specific immunoaffinity method to enrich blood NDEVs for biomarker development.

Methods: After screening multiple neuronal antigens, we achieved NDEV capture with high affinity and specificity using antibodies against Growth-Associated Protein (GAP) 43 and Neuroligin 3 (NLGN3). The EV identity of the captured material was confirmed by electron microscopy, western blotting, and proteomics. The specificity for neuronal origin was demonstrated by showing enrichment for neuronal markers (proteins, mRNA) and recovery of spiked neuronal EVs. We performed NDEV isolation retrospectively from plasma samples from two cohorts of early AD patients (N = 19 and N = 40) and controls (N = 20 and N = 19) and measured p181-Tau, amyloid-beta (Aβ) 42, brain-derived neurotrophic factor (BDNF), precursor brain-derived neurotrophic factor (proBDNF), glutamate receptor 2 (GluR2), postsynaptic density protein (PSD) 95, GAP43, and syntaxin-1.

Results: p181-Tau, Aβ42, and NRGN were elevated in AD samples, whereas proBDNF, GluR2, PSD95, GAP43, and Syntaxin-1 were reduced. Differences for p181-Tau, proBDNF, and GluR2 survived multiple-comparison correction and were correlated with cognitive scores. A model incorporating biomarkers correctly classified 94.7% of AD participants and 61.5% of control participants. The observed differences in NDEVs-associated biomarkers are consistent with previous findings.

Conclusion: NDEV isolation by GAP43 and NLGN3 immunocapture offers a robust novel platform for biomarker development in AD, suitable for large-scale validation.

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