Weiss Alexander, Meissner Alex Florin, Boros Fanni Annamária, Regensburger Martin, Taudte Regina Verena, Matamoros-Angles Andreu, Arnold Philipp, Zunke Friederike
Department of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander-University Erlangen-Nürnberg (FAU), Erlangen, Germany.
Institute of Functional and Clinical Anatomy, Friedrich-Alexander-University Erlangen-Nürnberg, Erlangen, Germany.
Cell Commun Signal. 2025 Sep 18;23(1):395. doi: 10.1186/s12964-025-02383-w.
Parkinson's disease is a neurodegenerative disorder with no curative treatment option and objective biomarker profile. Extracellular Vesicles (EVs) are membrane-enclosed biological nanoparticles released from all cells of the human body. In this pilot study we compare plasma- and serum-derived EVs from Parkinson's disease (PD) patients and healthy controls (HC) utilizing a precipitation-based method. Additionally, we employ an L1CAM antibody to selectively enrich for L1CAM-positive EVs from the total plasma-/serum-derived EV fractions. Successful EV enrichment was shown in western blot experiments for CD63 and for L1CAM as well as in metabolomic analysis for a HC sample. In a side-by-side quantification,. which we based on transmission electron microscopic images from negative stain samples, we identify small but significant differences between EV diameter from PD patients and HC. To streamline the quantification process, we introduce an ImageJ-based computer algorithm for (semi-)automated quantification of EVs from negative stain electron micrographs. We observe that this (semi-)automated quantification determines a smaller diameter than manual quantification. However, the difference between PD and HC group is systematic and reveals the same relative differences calculated from manually measured total plasma-derived EV particles. In this pilot study, we introduce a new workflow implemented into an ImageJ plugin enabling to determine differences in EV size within TEM images. For our data set of plasma-derived EVs from PD patients and HC, we find small, yet consistent differences. We feel that this study contributes to the search of a clinical biomarker for PD.
帕金森病是一种神经退行性疾病，目前尚无治愈性治疗方案和客观的生物标志物特征。细胞外囊泡（EVs）是从人体所有细胞释放的膜包裹生物纳米颗粒。在这项初步研究中，我们使用基于沉淀的方法比较了帕金森病（PD）患者和健康对照（HC）的血浆和血清来源的EVs。此外，我们使用L1CAM抗体从总的血浆/血清来源的EVs组分中选择性富集L1CAM阳性的EVs。在针对CD63和L1CAM的蛋白质印迹实验以及针对一个HC样本的代谢组学分析中，均显示出成功的EVs富集。在一项基于负染样本的透射电子显微镜图像的并列定量分析中，我们确定了PD患者和HC的EVs直径之间存在微小但显著的差异。为了简化定量过程，我们引入了一种基于ImageJ的计算机算法，用于对负染电子显微镜图像中的EVs进行（半）自动定量。我们观察到，这种（半）自动定量所确定的直径比手动定量小。然而，PD组和HC组之间的差异是系统性的，并且揭示了从手动测量的总的血浆来源的EV颗粒计算得出的相同相对差异。在这项初步研究中，我们引入了一种新的工作流程，该流程已在一个ImageJ插件中实现，能够确定透射电子显微镜图像中EVs大小的差异。对于我们来自PD患者和HC的血浆来源的EVs数据集，我们发现了微小但一致的差异。我们认为这项研究有助于寻找帕金森病的临床生物标志物。
