An acoustofluidic device for diagnosing preclinical Alzheimer's disease

摘要 阿尔茨海默病(AD)是一种残废的终末性神经退行性疾病，估计有550万人受到影响 美国人口和大约10%的65岁以上人口。的早期症状。 疾病包括近期记忆障碍、语言表达困难和行为改变 包括情绪低落。随着时间的推移，症状逐渐恶化，最终导致痴呆症和 身体机能。目前尚无灵敏、客观的诊断试验用于AD的早期诊断，使得 医生很难正确筛查这种疾病，也无法诊断出可能呈阳性的患者 广告。虽然AD的医学成像已经取得了进展，但它们极其昂贵，而且有限 诊断准确率。近年来，神经元来源的外体(30-150 nm的胞外小泡)已经 被认为是诊断阿尔茨海默病的一个有前途的生物标志物。在循环中发现的神经元来源的外切体可以提供 这是一种简单、非侵入性的中枢神经系统健康监测方法。积累证据 提示神经元衍生的外切体可能通过帮助传播，在AD的病理中发挥关键作用 异常的、潜在致病的、错误折叠的蛋白质遍布整个大脑。初步研究表明 通过分析神经元衍生的外切体及其分子货物的数量，早期AD患者 可以与健康对照组以及其他神经疾病患者区分开来。而当 研究人员在鉴定神经元来源的外体蛋白和RNA方面取得了进展，难度很大 围绕外切体的分离和分析，阻止了它们作为AD生物标记物的广泛使用。 目前，还没有商业化的产品能够同时分离和分析 神经元衍生的外切体。该SBIR项目的目标是克服现有AD的局限性 诊断技术，并通过开发和商业化解决市场上未得到满足的需求 自动化、高纯度、高产量、生物相容的外切体分离和准确的AD生物标志物检测 声流(即声学和流体力学的融合)分离和电化学检测 技术。在第一阶段项目的工作中，我们成功地展示了 研制的满足或超过目标值诊断AD的外切体分离分析装置 对于成功衡量标准中确定的四个关键参数中的每一个。在第二阶段，我们的商业化 活动将提高声流控芯片的性能，开发自给自足的测试版 原型，并与最终用户验证其性能。凭借其在自动化、速度、精度、 和准确性，建议的声流控技术有可能极大地简化和革命性地 诊断阿尔茨海默病。结果将是一个高度敏感的液体活检，它将提供一个全面的分子 AD的签名，从而显著提高了诊断准确性，超越了现有方法，并提供了更深入的 对疾病的病理生理学有深入的了解。

Abstract Alzheimer’s disease (AD) is a crippling, terminal neurodegenerative illness that affects an estimated 5.5 million people in the United States and approximately 10% of the population over the age of 65. Early symptoms of the disease include impairment in recent memory, difficulties with expressive language, and behavioral changes including depressed mood. Symptoms gradually worsen over time, ultimately leading to dementia and a loss of bodily functions. Currently there is no sensitive, objective diagnostic test for the early diagnosis of AD, making it difficult for physicians to properly screen for the disease and failing to diagnose patients that may be positive for AD. While advances have been made in medical imaging for AD, they are extremely costly and have limited diagnostic accuracy. In recent years, neuron-derived exosomes (30-150 nm extracellular vesicles) have emerged as a promising biomarker for diagnosing AD. Neuron-derived exosomes found in circulation can provide a simple, non-invasive means of monitoring the health of the central nervous system. Accumulating evidence suggests that neuron-derived exosomes may play a crucial role in the pathology of AD by helping to spread abnormal, potentially disease-causing, misfolded proteins throughout the brain. Preliminary studies have shown that by analyzing the number of neuron-derived exosomes and their molecular cargo, early-stage AD patients can be distinguished from healthy controls, as well as patients with other neurological diseases. While researchers have made progress in identifying neuron-derived exosomal proteins and RNAs, difficulties surrounding the isolation and analysis of exosomes have prevented their widespread use as a biomarker for AD. Currently, there are no commercially available products capable of simultaneously isolating and analyzing neuron-derived exosomes. The objective of this SBIR project is to overcome the limitations of existing AD diagnostic technologies and address the unmet needs in the market by developing and commercializing an automated, high-purity, high-yield, biocompatible exosome isolation and accurate AD biomarker detection using acoustofluidic (i.e., the fusion of acoustics and fluid mechanics) separation and electrochemical detection technologies. During our work on the Phase I project, we successfully demonstrated the utility and feasibility of the proposed exosome isolation and analysis device for AD diagnosis by meeting or exceeding the target values for each of the four key parameters identified in the Measures of Success. In Phase II, our commercialization activities will improve the performance of the acoustofluidic chips, develop self-contained, beta-testing-ready prototypes, and validate their performance with end users. With its advantages in automation, speed, precision, and accuracy, the proposed acoustofluidic technology has the potential to greatly simplify and revolutionize the diagnosis of AD. The result will be a highly sensitive liquid biopsy that will provide a comprehensive molecular signature for AD, thus significantly improving diagnostic accuracy past existing methods and providing deeper insight in the pathophysiology of the disease.