单细胞外囊泡 (EV) 分析验证了 L1 细胞粘附分子 (L1CAM) 是神经元衍生 EV 的可靠生物标记。

IF 15.5 1区 医学 Q1 CELL BIOLOGY Journal of Extracellular Vesicles Pub Date : 2024-06-13 DOI:10.1002/jev2.12459
Carlos J Nogueras-Ortiz, Erden Eren, Pamela Yao, Elizabeth Calzada, Christopher Dunn, Olga Volpert, Francheska Delgado-Peraza, Maja Mustapic, Alexey Lyashkov, F Javier Rubio, Michael Vreones, Lesley Cheng, Yang You, Andrew F Hill, Tsuneya Ikezu, Erez Eitan, Edward J Goetzl, Dimitrios Kapogiannis
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引用次数: 0

摘要

用 L1 细胞粘附分子(L1CAM)特异性抗体分离神经元衍生的细胞外囊泡(NDEVs)已被广泛用于鉴定中枢神经系统疾病的血液生物标记物。然而,方法论的全面验证需要在单个 NDEV 中证明 L1CAM,以及在来自其他细胞的单个 EV 中证明 L1CAM 水平较低或不存在。在这里,我们使用多种单个 EV 技术确定了神经元的来源,并确定了人体血液中 L1CAM 阳性 EV 的丰度。结果表明,L1CAM 外结构域的表位与神经元蛋白 β-III-tubulin、GAP43 和 VAMP2 共同表达在单个 EV 上,它们的水平随着 L1CAM 阳性 EV 的富集而增加。携带神经元蛋白 VAMP2 和 β-III-tubulin 的 L1CAM 阳性 EVs 含量为 30% 至 63%,而 L1CAM 阴性 EVs 含量为 0.8% 至 3.9%。血浆液相 L1CAM 不与单个 EV 结合。我们的研究结果支持将 L1CAM 作为分离血浆 NDEVs 的目标,并利用它们的货物来鉴定反映神经元功能的生物标记物。
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Single-extracellular vesicle (EV) analyses validate the use of L1 Cell Adhesion Molecule (L1CAM) as a reliable biomarker of neuron-derived EVs

Isolation of neuron-derived extracellular vesicles (NDEVs) with L1 Cell Adhesion Molecule (L1CAM)-specific antibodies has been widely used to identify blood biomarkers of CNS disorders. However, full methodological validation requires demonstration of L1CAM in individual NDEVs and lower levels or absence of L1CAM in individual EVs from other cells. Here, we used multiple single-EV techniques to establish the neuronal origin and determine the abundance of L1CAM-positive EVs in human blood. L1CAM epitopes of the ectodomain are shown to be co-expressed on single-EVs with the neuronal proteins β-III-tubulin, GAP43, and VAMP2, the levels of which increase in parallel with the enrichment of L1CAM-positive EVs. Levels of L1CAM-positive EVs carrying the neuronal proteins VAMP2 and β-III-tubulin range from 30% to 63%, in contrast to 0.8%–3.9% of L1CAM-negative EVs. Plasma fluid-phase L1CAM does not bind to single-EVs. Our findings support the use of L1CAM as a target for isolating plasma NDEVs and leveraging their cargo to identify biomarkers reflecting neuronal function.

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来源期刊
Journal of Extracellular Vesicles
Journal of Extracellular Vesicles Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
27.30
自引率
4.40%
发文量
115
审稿时长
12 weeks
期刊介绍: The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies. The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.
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