Département de pharmacologie et physiologie, Institut de Génie Biomédical, Faculté de Médecine, Université de Montréal, Montréal, Quebec, Canada.
Multiomics Investigation of Neurodegenerative Diseases (MIND) lab, Centre de recherche de l'Institut Universitaire de Gériatrie, Montréal, Quebec, Canada.
Alzheimers Dement. 2024 Jul;20(7):4411-4422. doi: 10.1002/alz.13823. Epub 2024 Jun 12.
Brain-derived extracellular vesicles (BEVs) in blood allows for minimally-invasive investigations of central nervous system (CNS) -specific markers of age-related neurodegenerative diseases (NDDs). Polymer-based EV- and immunoprecipitation (IP)-based BEV-enrichment protocols from blood have gained popularity. We systematically investigated protocol consistency across studies, and determined CNS-specificity of proteins associated with these protocols.
NDD articles investigating BEVs in blood using polymer-based and/or IP-based BEV enrichment protocols were systematically identified, and protocols compared. Proteins used for BEV-enrichment and/or post-enrichment were assessed for CNS- and brain-cell-type-specificity, extracellular domains (ECD+), and presence in EV-databases.
A total of 82.1% of studies used polymer-based (ExoQuick) EV-enrichment, and 92.3% used L1CAM for IP-based BEV-enrichment. Centrifugation times differed across studies. A total of 26.8% of 82 proteins systematically identified were CNS-specific: 50% ECD+, 77.3% were listed in EV-databases.
We identified protocol steps requiring standardization, and recommend additional CNS-specific proteins that can be used for BEV-enrichment or as BEV-biomarkers.
Across NDDs, we identified protocols commonly used for EV/BEV enrichment from blood. We identified protocol steps showing variability that require harmonization. We assessed CNS-specificity of proteins used for BEV-enrichment or found in BEV cargo. CNS-specific EV proteins with ECD+ or without were identified. We recommend evaluation of blood-BEV enrichment using these additional ECD+ proteins.
血液中的脑源性细胞外囊泡 (BEV) 可实现对中枢神经系统 (CNS) 特异性标志物的微创研究，这些标志物与年龄相关的神经退行性疾病 (NDD) 相关。基于聚合物的 EV 和免疫沉淀 (IP) 富集 BEV 的方案在血液中得到了广泛应用。我们系统地研究了不同研究中方案的一致性，并确定了与这些方案相关的蛋白质的 CNS 特异性。
系统地鉴定了使用基于聚合物和/或基于 IP 的 BEV 富集方案研究血液中 BEV 的 NDD 文章，并对方案进行了比较。评估了用于 BEV 富集和/或富集后蛋白质的 CNS 和脑细胞特异性、细胞外结构域 (ECD+) 和 EV 数据库中的存在情况。
共有 82.1%的研究使用聚合物 (ExoQuick) EV 富集，92.3%的研究使用 L1CAM 进行基于 IP 的 BEV 富集。研究中离心时间不同。在系统鉴定的 82 种蛋白质中，共有 26.8%是 CNS 特异性的：50% ECD+，77.3%列在 EV 数据库中。
我们确定了需要标准化的方案步骤，并推荐了可用于 BEV 富集或作为 BEV 生物标志物的其他 CNS 特异性蛋白质。
在 NDD 中，我们确定了常用于从血液中富集 EV/BEV 的方案。我们确定了需要协调的显示变异性的方案步骤。我们评估了用于 BEV 富集或在 BEV 货物中发现的蛋白质的 CNS 特异性。鉴定了具有 ECD+或无 ECD+的 CNS 特异性 EV 蛋白质。我们建议使用这些额外的 ECD+蛋白质评估血液 BEV 富集。
