CAREER: Ultrafast molecular separation and integrated near-field light-metal-fluorophore interactions for biomarker detection at point-of-care

癌症、肥胖和阿片类药物滥用对美国人民的健康构成了综合威胁，影响了很大一部分人的健康，每年的医疗费用超过2500亿美元。解决与疾病起始、进展和治疗干预相关的基本分子变化的传感技术的不可用性是成功对抗这些疾病的关键障碍。microRNA（miRNAs）是一类短的非编码RNA片段（约17-25个核苷酸），通过降解或抑制多个靶mRNA的翻译参与基因表达的转录后调控。由于miRNA参与启动许多疾病的生化级联反应的早期基因调控，因此它们可以比其他下游生物标志物（例如，抗原）。此外，miRNA生物标志物在循环血液中稳定表达，因此，miRNA检测可用于开发灵敏、微创和低成本的诊断测试。不幸的是，miRNAs尚未被翻译或用于任何疾病的临床诊断。这部分是由于缺乏适用于诊所、医院和医疗中心的灵敏且低成本的miRNA检测方法。这项研究计划的成果将是一个完整的，随时可用的miRNA检测平台，包括数据收集，质量控制和内置的数据分析统计方法。此外，拟议的更广泛的影响活动旨在将研究引入北达科他州州立大学的本科电气工程课程。此外，外联工作的目标是通过引入生物传感器概念和提供机会，为社会或社区问题提供技术解决方案，吸引代表性不足的群体（中学女生和美洲土著社区大学生）进入工程领域。PI还将与美国土著学校的高中STEM教师合作，帮助他们为学生制定学习计划。这些活动将扩大代表性不足的群体在STEM教育和劳动力中的参与。本计画的研究目标是探讨小核酸场领域的三个基本概念（电和温度）相互作用和近场光-金属-荧光团能量转移：（1）核酸基础与领域研究（电和温度）相互作用，以选择性地分离荧光团标记的短（~17-25个核苷酸）miRNA-DNA双链体分子，并将它们集中在电极附近，（2）研究AC电场以使有助于增强荧光团的荧光强度的每个近场光-金属-荧光团相互作用最大化，和3）整合多种相互作用以增强荧光强度，使其远远超过当前的能力。这一新知识将用于开发通用生物传感器的模板，该生物传感器可检测任何缓冲液中的任何生物标志物类型的个体或组合。具体而言，拟议的5年项目将开发基于miRNA的床旁技术，以满足临床样本分析的迫切需求。该项目由电气，通信和网络系统部门（ECCS）共同资助，激励竞争研究计划（EPSCoR）该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Cancer, obesity, and opioid abuse pose a combined threat to the health of the people in the United States, affecting a significant portion and costing more than $250 billion per year in medical expenses. The unavailability of sensing technologies addressing the fundamental molecular changes related to disease initiation, progression, and therapeutic interventions is a critical roadblock for successfully combating these diseases. MicroRNA (miRNAs) are short non-coding RNA fragments (~17–25 nucleotides) that are involved in post-transcriptional regulation of gene expression either by degrading or repressing translation of multiple target mRNAs. Since miRNAs are involved in early gene regulations that initiate biochemical cascades for many diseases, they could provide reliable and clinically relevant information earlier than other downstream biomarkers (e.g., antigens). Also, miRNA biomarkers are stably expressed in circulating blood and, therefore, miRNA detection could be used to develop sensitive, minimally invasive, and low-cost diagnostics tests. Unfortunately, miRNAs have not yet been translated or utilized in the clinical diagnosis of any disease. This is in part due to the lack of sensitive and low-cost miRNA detection approaches that are applicable at clinics, hospitals, and medical centers. The outcome of this research plan will be a complete, ready-to-go miRNA detection platform that includes data collection, quality control, and built-in statistical methods for data analysis. In addition, proposed broader impact activities aim to introduce research into the undergraduate electrical engineering curriculum of North Dakota State University. Also, outreach efforts will aim to attract underrepresented groups (middle school girls and Native American community college students) to the field of engineering by introducing biosensor concepts and providing an opportunity to produce a technical solution to a societal or community problem. The PI will also work with high school STEM teachers in Native American schools to help them develop learning plans for their students. These activities will broaden the participation of underrepresented groups in STEM education and the workforce. The research goals of this project are to study three fundamental concepts in the areas of small nucleic acid-field (electric and temperature) interactions and near-field light-to-metal-to-fluorophore energy transfer: (1) study of fundamental nucleic acid and field (electric and temperature) interactions to selectively separate fluorophore-labeled short (~17–25 nucleotide) miRNA-DNA duplex molecules from other single-stranded nucleic acid molecules of similar lengths and concentrate them near electrodes, (2) study of AC electric fields to maximize each near-field light-metal-fluorophore interaction that contributes to enhancing the fluorescence intensity of fluorophores, and 3) integration of multiple interactions to enhance the fluorescence intensity well beyond current capabilities. This new knowledge will be used to develop a template for a universal biosensor that detects any individual or combination of biomarker-types in any buffer. Specifically, the proposed 5-year project will develop miRNA-based point-of-care technology to address the critical need for the analysis of clinical samples.This project is jointly funded by the Electrical, Communications and Cyber Systems Division (ECCS), and the Established Program to Stimulate Competitive Research (EPSCoR).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.