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

该项目的主要目的是实现电化学微流体多路复用芯片实验室（MEMS）平台，用于对人体体液中一组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，开发的系统将被应用于分析相关的生物标本的阿尔茨海默病患者，主要是人的脑脊液（CSF）和外周血。在阿尔茨海默病的临床诊断，最流行的形式的痴呆症，有一个伟大的和迫切需要的新的和互补的生物标志物，以监测病理过程，包括神经递质消耗或神经炎症。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进行适当的诊断和治疗的监测，因此非常希望利用快速、低成本和可靠的系统同时沿着测量不同的生物标志物。因此，该项目致力于使用具有用于即时检测（POCT）的多路复用微流体的电化学生物传感器平台来实现神经递质以及与神经炎症相关的一组miRNA的新型检测策略。P物质不仅作为神经元信号传递物起作用，而且作为由外周和中枢神经系统免疫细胞表达炎性介质起作用。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学中的参与及其作为临床生物标志物的可用性都没有得到充分的澄清。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。MicroRNA，小的非编码RNA分子，在基因表达的调节中起着关键作用。一些miRNAs是疾病特异性表达的，可用于生物标志物分析。因此，随着许多筛选研究旨在鉴定癌症、心血管疾病、痴呆症（如AD）和神经炎症的新的潜在生物标志物候选物，miRNA变得越来越重要。在本项目的第二部分中，我们将对各种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研究，以实现和测试用于临床诊断的4重miRNA面板。这样的平台将成为阿尔茨海默病研究及其他领域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实现。

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.