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

项目概要 阿尔茨海默病（AD）是一种复杂且严重的神经退行性疾病。 AD的特点是 随着老年斑（含有蛋白质的沉积物）的积累，大脑中的突触和神经元丧失， 神经原纤维缠结。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费用估计为 2260亿美元。然而，目前还没有明确的 AD 即时护理 (POC) 诊断方法。当前AD的标准方法 诊断涉及影像学和认知测试的结合，这些测试昂贵且需要复杂的、 耗时的实验室分析，而明确的诊断是在事后进行的。因为早 诊断可以实现更好的计划和协调护理，迫切需要开发一种 用于症状前诊断的经济高效、高度敏感和选择性的诊断工具，也可以 可用作床边或主要 POC 使用的便捷监测设备。 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) 以及患者的脑脊液样本。 ISL 证明 Adnos 设备的平均限制为 检测 100 fM AD 特异性生物标志物。结果表明，Adnos 的灵敏度优于 标准实验室测试，例如 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 检测性能 AD 特异性生物标志物。 ISL 将：(1) 优化 Adnos 纳米线传感器制造工艺，(2) 进行检测 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 在不到 30 分钟的时间内，(4) 展示了原型 Adnos 检测临床脑脊液中 AD 生物标志物的能力 样品并与标准 ELISA 和斑点印迹测试进行比较，以及 (5) 准备确保 CLIA 在我们探索商业选择时放弃。 在症状出现之前检测 AD 最早阶段的能力可能具有强大的作用 对家庭的影响：(1) 规划未来的医疗保健需求，(2) 制定早期干预措施 策略，(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将非常有价值。 AD 的有效治疗方法。

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.