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共同威胁，影响了很大一部分人，每年的医疗费用超过2500亿美元。传感技术无法解决与疾病发生、进展和治疗干预相关的基本分子变化，这是成功对抗这些疾病的关键障碍。MicroRNA （mirna）是一种短的非编码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参与了许多疾病的早期基因调控，启动了生化级联反应，因此它们可以比其他下游生物标志物（如抗原）更早地提供可靠的临床相关信息。此外，miRNA生物标志物在循环血液中稳定表达，因此，miRNA检测可用于开发敏感、微创和低成本的诊断测试。不幸的是，mirna尚未被翻译或用于任何疾病的临床诊断。这在一定程度上是由于缺乏适用于诊所、医院和医疗中心的敏感和低成本的miRNA检测方法。这项研究计划的结果将是一个完整的，现成的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，提议的更广泛的影响活动旨在将研究引入北达科他州立大学的本科电气工程课程。此外，通过引入生物传感器概念，并提供机会为社会或社区问题提供技术解决方案，外展工作将旨在吸引代表性不足的群体（中学女生和美国原住民社区大学生）进入工程领域。PI还将与美国原住民学校的高中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对STEM教育和劳动力的参与。本项目的研究目标是研究小核酸场（电和温度）相互作用和近场光-金属-荧光团能量转移领域的三个基本概念：(1)研究基本的核酸和场（电和温度）相互作用，选择性地将荧光团标记的短（~ 17-25个核苷酸）miRNA-DNA双分子与其他类似长度的单链核酸分子分离，并将其集中在电极附近；(2)研究交流电场，最大限度地提高每个近场轻金属-荧光团相互作用，有助于增强荧光团的荧光强度；3)整合多种相互作用，提高荧光强度，远远超出目前的能力。这一新知识将用于开发通用生物传感器模板，用于检测任何缓冲液中的任何个体或生物标志物类型的组合。具体来说，拟议的5年项目将开发基于mirna的即时护理技术，以解决临床样本分析的关键需求。该项目由电气、通信和网络系统部（ECCS）和促进竞争研究的既定计划（EPSCoR）共同资助。该奖项反映了美国国家科学基金会的法定使命，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.