Nanowire Sensor Array-based Assay for Early Diagnosis of Alzheimer's Disease

项目摘要 阿尔茨海默病（Alzheimer's disease，AD）是一种复杂而严重的神经退行性疾病。AD的特征在于 大脑中的突触和神经元损失，伴有老年斑（含蛋白质的沉积物）的积累， 神经系统缠结大约540万美国人和全球3000万人受到影响， 数量正在迅速增长。2015年，AD和其他痴呆症的累计护理成本估计为 2260亿美元然而，AD没有明确的床旁（POC）诊断。AD的现行标准方法 诊断涉及昂贵且需要复杂的成像和认知测试的组合， 耗时的实验室分析，而明确的诊断是在死后进行的。因为早期 诊断可以更好地规划和协调护理，迫切需要制定一个 具有成本效益，高度敏感和选择性的诊断工具，用于症状前AD诊断，还可以 可用作床边或主要POC使用的方便监测装置。 在第一阶段，InnoSense LLC（ISL）开发了一种基于导电聚合物纳米线的生物传感器Adnos，用于 检测AD生物标志物。ISL构建了一个工作模型，并展示了其检测能力 磷酸盐缓冲盐水和人工脑脊液加标溶液中的AD相关生物标志物 （aCSF）以及来自患者的CSF样品。ISL证明Adnos设备的平均限值为 AD特异性生物标志物的100 fM检测。结果表明，Adnos具有比 标准实验室测试，例如用于AD蛋白的酶联免疫吸附测定（ELISA）（纳摩尔 分析. 在第二阶段，ISL将进一步开发、微调和严格评估Adnos的性能， AD特异性生物标志物。ISL将：（1）优化Adnos纳米线传感器的制造工艺，（2） 开发用必要的电子设备和软件构建原型，（3）微调原型 准确检测和监测AD特异性生物标志物的性能，检测限为100 fM 在少于30分钟内，（4）证明原型Adnos检测临床CSF中AD生物标志物的能力 样品，并与标准ELISA和斑点印迹试验的性能进行比较，以及（5）准备确保CLIA 我们会放弃豁免权同时探索商业选择 在症状发作之前检测AD的最早阶段的能力可能具有强大的 对家庭的影响：（1）规划未来的保健需求，（2）制定早期干预措施 策略，以及（3）获得理解和管理疾病整个生命周期的能力。使用 Adnos系统作为一种为临床研究提供监测数据的工具， 有效治疗AD。

PROJECT SUMMARY Alzheimer’s disease (AD) is a complex and severe neurodegenerative disorder. AD is characterized by synapse and neuron loss in the brain with the accumulation of senile plaques (protein containing deposits) and neurofibrillary tangles. Approximately 5.4 million Americans and globally 30 million people are affected and this number is growing rapidly. In 2015, the cumulative cost of care for AD and other dementia was estimated at $226 billion. Yet, there is no definitive point-of-care (POC) diagnostic for AD. Current standard methods of AD diagnosis involve a combination of imaging and cognitive tests which are expensive and require complicated, time-consuming laboratory-based analysis while definitive diagnosis is made postmortem. Because early diagnosis could enable better planned and coordinated care, there is an urgent need to develop a cost-effective, highly sensitive and selective diagnostic tool for pre-symptomatic AD diagnosis which could also be used as a convenient monitoring device for bedside or primary POC use. In Phase I, InnoSense LLC (ISL) developed a conducting polymer nanowire-based biosensor, Adnos, for detecting AD biomarkers. ISL constructed a working model and demonstrated its capability for detecting AD-associated biomarkers in spiked solutions with phosphate buffered saline and artificial cerebrospinal fluid (aCSF) as well as CSF samples from patients. ISL demonstrated that Adnos devices had an average limit of detection of 100 fM for AD-specific biomarkers. The results indicate that Adnos has better sensitivity than standard laboratory tests such as Enzyme-Linked Immunosorbent Assay (ELISA) (nanomolar) for AD protein analysis. In Phase II, ISL will further develop, fine tune, and rigorously evaluate Adnos performance for detecting AD-specific biomarkers. ISL will: (1) optimize the Adnos nanowire sensor fabrication process, (2) toward assay development construct a prototype with necessary electronics and software, (3) fine-tune the prototype performance for accurate detection and monitoring of AD-specific biomarkers with a limit of detection of 100 fM in less than 30 min, (4) demonstrate ability of a prototype Adnos to detect AD biomarkers in clinical CSF samples and compare the performance with standard ELISA and dot blot tests, and (5) prepare to secure CLIA waiver while we explore commercial options. The ability to detect the earliest stages of AD, prior to onset of symptoms, could potentially have a powerful impact on families to: (1) plan for future healthcare needs, (2) development of early stage intervention strategies, and (3) gain the ability to understand and manage the entire lifecycle of the disease. Use of the Adnos system will be highly valuable as a tool providing monitoring data for clinical studies in search of effective treatments for AD.