Multiplexed point-of-care testing for inflammatory miRNA and neurotransmitter profiling in Alzheimer’s disease diagnostics

该项目的主要目标是实现一个电化学微流体多路芯片实验室（LOC）平台，用于对人体体液中潜在的阿尔茨海默病（AD）生物标志物进行敏感和现场测试。在这方面，神经炎症过程的各种生物标志物，包括神经递质和microrna，将被测量和评估其在临床诊断中的适用性。为此，开发的系统将用于分析阿尔茨海默病患者的相关生物标本，主要是人脑脊液（CSF）和外周血。在阿尔茨海默病的临床诊断中，迫切需要新的和互补的生物标志物来监测包括神经递质耗竭或神经炎症在内的病理过程。由于在许多情况下，基于单一生物标志物的临床发现不足以对治疗进行适当的诊断和监测，因此非常需要同时测量不同的生物标志物以及快速，低成本和可靠的系统。因此，该项目致力于实现一种新的神经递质检测策略，以及一组与神经炎症相关的mirna，使用电化学生物传感器平台与多路微流体进行即时检测（POCT）。神经递质，如P物质，不仅作为神经元的信号递质，而且作为炎症介质由外周和中枢神经系统免疫细胞表达。尽管对神经递质在阿尔茨海默病中的作用进行了大量的研究，但这种多功能递质在疾病病理中的作用及其作为临床生物标志物的可用性都没有得到充分的阐明。因此，该项目将特别有助于阐明神经递质作为AD生物标志物的适用性。MicroRNAs是一种小的非编码RNA分子，在基因表达调控中起着关键作用。一些mirna是疾病特异性表达的，可以用于生物标志物分析。因此，随着众多筛选研究旨在识别癌症、心血管疾病、痴呆（如AD）和神经炎症等新的潜在生物标志物候选物，mirna正变得越来越重要。在该项目的第二部分，将对各种miRNA作为阿尔茨海默病潜在生物标志物候选物的临床验证进行检查，以实施和测试用于临床诊断的4-plex miRNA小组。提出的微流体POCT装置的成功演示将提供许多好处，例如直接和快速检测mirna，而无需任何事先扩增。这样一个平台将是阿尔茨海默病研究中神经炎症分析的一个重要里程碑，因为测量原理可以在许多应用领域得到实施。

The major aim of this project is the implementation of an electrochemical micro-fluidic multiplexed lab-on-a-chip (LOC) platform for the sensitive and on-site testing of a panel of potential Alzheimer’s disease (AD) biomarkers in human body fluids. In this regard, various biomarkers of neuroinflammatory processes, including neurotransmitters and microRNAs, will be measured and evaluated for their applicability in clinical diagnostics. For this purpose, the developed system will be applied for the analysis of relevant biospecimen of Alzheimer patients, primarily human cerebrospinal fluid (CSF) and peripheral blood.In the clinical diagnostics of Alzheimer’s disease, the most prevalent form of dementia, there is a great and urgent need for novel and complementary biomarkers to monitor pathological processes including neurotransmitter depletion or neuroinflammation. As in many cases clinical findings on the basis of a single biomarker are not sufficient for the appropriate diagnosis and surveillance of the therapy, it is highly desirable to gauge different biomarkers simultaneously along with a rapid, low-cost and reliable system. Thus, this project is dedicated to implement a novel detection strategy for the neurotransmitters as well as a panel of miRNAs associated with neuroinflammation using an electrochemical biosensor platform with multiplexed microfluidics for point-of-care testing (POCT).The neurotransmitters, e.g., substance P, not only function as a neuronal signal transmitter, but also as an inflammatory mediator expressed by both peripheral and central nervous system immune cells. Despite a significant amount of research on the role of neurotransmitters in Alzheimer’s disease, neither the involvement of this multifunctional transmitter in the disease pathology nor its usability as a clinical biomarker have been sufficiently clarified. Hence, this project will be especially useful for clarifying the applicability of neurotransmitters as a biomarker in AD.MicroRNAs, small non-coding RNA molecules, play a key role in the regulation of gene expression. Some miRNAs are expressed disease-specific and can utilized for biomarker analysis. Hence, miRNAs are gaining increasing importance as numerous screening studies aim to identify new potential biomarker candidates for cancer, cardiovascular disease, dementias such as AD and neuroinflammation. In the second part of this project, the clinical validation of various miRNAs as potential biomarker candidates for Alzheimer’s disease will be examined to implement and test a 4-plex miRNA panel for clinical diagnosis.A successful demonstration of the proposed microfluidic POCT device will offer many benefits such as direct and rapid detection of miRNAs without any prior amplification. Such a platform would be a major milestone on neuroinflammation analysis in Alzheimer research and beyond as the principle of measurement could be implemented on many areas of application.