Jean M Lewis, Dorathy-Ann Harris, Janell Kosmatka, Emily Mikrut, Jack Evenson, Heath I Balcer, Harmeet Dhani, Juan Pablo Hinestrosa, Robert Rissman, Paul R Billings
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引用次数: 0
Abstract
Background: Blood tests for Alzheimer's disease (AD) that measure biomarkers related to neuropathology have demonstrated to be useful, minimally-invasive ways to identify patients for screening into clinical trials. While some AD biomarkers can be detected in plasma, greater sensitivity is needed to make plasma AD tests more effective. Extracellular vesicles (EVs) in plasma carry AD-related biomarkers from the brain and could offer a concentrated source of brain-related biomarkers, though the methodological complexities involved in isolating plasma EVs have hampered its validation for clinical use.
Objective: To explore the feasibility and effectiveness of developing blood tests for AD utilizing extracellular vesicle-bound protein biomarkers.
Methods: We developed a simplified method for isolating EVs directly from plasma using an alternating current electrokinetic (ACE) microchip. No sample pretreatment steps were needed. Protein biomarkers on the EVs were detected by adding fluorescent antibodies to the plasma samples before capture by the chip. This allowed measurement of EV biomarker levels directly on the chip.
Results: AD or non-AD control plasma was measured for ten different AD-related biomarkers. EV-associated NCAM1, pTau231, α-synuclein, and TDP-43 levels were able to distinguish a group of 10 AD, 10 mild cognitive impairment (MCI), and 10 non-AD subjects. pTau231 was different between AD and non-AD (p = 0.0300) and α-synuclein differentiated AD from MCI (p = 0.0148).
Conclusions: This study shows how ACE microfluidic chip technology can help differentiate AD and MCI patients from non-AD controls with clinical relevance. This work also highlights the important diagnostic role of plasma EV biomarkers in neurodegenerative disease.
背景：对阿尔茨海默病（AD）进行血液检测，测量与神经病理学相关的生物标志物，已被证明是一种有用的、微创的方法，可用于筛选患者进入临床试验。虽然血浆中可以检测到某些阿兹海默症生物标志物，但要使血浆阿兹海默症检测更有效，还需要更高的灵敏度。血浆中的细胞外囊泡（EVs）携带来自大脑的AD相关生物标志物，可以提供大脑相关生物标志物的集中来源，但分离血浆EVs的方法复杂，阻碍了其临床应用的验证：探索利用细胞外囊泡结合蛋白生物标记物开发AD血液检测的可行性和有效性：方法：我们开发了一种简化的方法，利用交变电流电动（ACE）微芯片直接从血浆中分离出EVs。无需样品预处理步骤。在芯片捕获血浆样品之前，通过向血浆中添加荧光抗体来检测EVs上的蛋白质生物标记物。这样就可以直接在芯片上测量EV生物标记物的水平：结果：对AD或非AD对照血浆进行了10种不同的AD相关生物标记物检测。EV相关的NCAM1、pTau231、α-突触核蛋白和TDP-43水平能够区分10名AD受试者、10名轻度认知障碍（MCI）受试者和10名非AD受试者。PTau231在AD和非AD之间存在差异（p = 0.0300），α-突触核蛋白区分了AD和MCI（p = 0.0148）：这项研究表明，ACE 微流控芯片技术有助于区分 AD 和 MCI 患者与非 AD 对照组，具有临床意义。这项工作还凸显了血浆 EV 生物标记物在神经退行性疾病中的重要诊断作用。
期刊介绍：
The Journal of Alzheimer''s Disease (JAD) is an international multidisciplinary journal to facilitate progress in understanding the etiology, pathogenesis, epidemiology, genetics, behavior, treatment and psychology of Alzheimer''s disease. The journal publishes research reports, reviews, short communications, hypotheses, ethics reviews, book reviews, and letters-to-the-editor. The journal is dedicated to providing an open forum for original research that will expedite our fundamental understanding of Alzheimer''s disease.
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