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

职业：超快分子分离和集成近场光-金属-荧光团相互作用，用于护理点生物标志物检测

• 批准号: 2310106
• 负责人: Dharmakeerthi Nawarathna
• 金额: $ 50万
• 依托单位: Old Dominion University Research Foundation
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2310106&HistoricalAwards=false
• 关键词: CAREER; Ultrafast; molecular; separation; integrated

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.

癌症、肥胖和阿片类药物滥用对美国人民的健康构成综合威胁，影响很大一部分，每年造成超过 2500 亿美元的医疗费用。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 的翻译来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）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。部分原因是缺乏适用于诊所、医院和医疗中心的灵敏且低成本的 miRNA 检测方法。该研究计划的成果将是一个完整的、随时可用的 miRNA 检测平台，其中包括数据收集、质量控制和用于数据分析的内置统计方法。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立大学的本科电气工程课程。此外，外展工作的目标是通过引入生物传感器概念并提供为社会或社区问题提供技术解决方案的机会，吸引代表性不足的群体（中学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。该项目的研究目标是研究小核酸场（电和温度）相互作用和近场光到金属到荧光团能量转移领域的三个基本概念：（1）研究基本核酸和场（电和温度）相互作用，以选择性地将荧光团标记的短（约17-25个核苷酸）miRNA-DNA双链体分子与其他单链核酸分离 类似长度的酸分子并将它们集中在电极附近，（2）研究交流电场以最大化每个近场光-金属-荧光团相互作用，这有助于增强荧光团的荧光强度，以及3）整合多种相互作用以增强荧光强度远远超出当前能力。这些新知识将用于开发通用生物传感器的模板，该模板可检测任何缓冲液中的任何个体或生物标记类型的组合。具体来说，拟议的为期 5 年的项目将开发基于 miRNA 的护理点技术，以满足临床样本分析的关键需求。该项目由电气、通信和网络系统部门 (ECCS) 以及刺激竞争性研究既定计划 (EPSCoR) 共同资助。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查进行评估，认为值得支持。 标准。

项目成果

Integrated Nano-Plasmonic and Dielectrophoretic Based Label Free DNA Biomarker Detection

基于集成纳米等离子体和介电泳的无标记 DNA 生物标志物检测

• DOI: 10.1109/rapid54473.2023.10264745
• 发表时间: 2023
• 期刊: IEEE
• 作者: Lakshan, K. A.;Nawarathna, Dharmakeerthi
• 通讯作者: Nawarathna, Dharmakeerthi

Label-free Biomarker Detection Using Dielectrophoresis and Localized Surface Plasmonic Resonance

使用介电泳和局部表面等离子共振进行无标记生物标志物检测

• DOI: 10.1109/rapid54472.2022.9911547
• 发表时间: 2022
• 期刊: IEEE RAPID
• 作者: Lakshan, K. A.;Nawarathna, Dharmakeerthi
• 通讯作者: Nawarathna, Dharmakeerthi

Label Free Flow Cytometry Using Machine Learning

使用机器学习进行无标签流式细胞术

• DOI: 10.1109/rapid54473.2023.10264775
• 发表时间: 2023
• 期刊: IEEE
• 作者: Vandal, Noah;Tida, Umamaheswa Rao;Nawarathna, Dharmakeerthi
• 通讯作者: Nawarathna, Dharmakeerthi

Concentration of Flurophore Labeled ssDNA Molecules Near Au Nanoparticles by Dielectrophresis Improves the Fluorescence Intensity

通过介电电泳将荧光团标记的 ssDNA 分子浓缩在 Au 纳米颗粒附近可提高荧光强度

• DOI: 10.1109/rapid54473.2023.10264718
• 发表时间: 2023
• 期刊: IEEE
• 作者: Lakshan, K. A.;Nawarathna, Dharmakeerthi
• 通讯作者: Nawarathna, Dharmakeerthi

A Miniaturized MicroRNA Sensor Identifies Targets Associated with Weight Loss in a Diet and Exercise Intervention among Healthy Overweight Individuals.

• DOI: 10.3390/s22186758
• 发表时间: 2022-09-07
• 期刊: Sensors (Basel, Switzerland)
• 作者: Jayasooriya V;Johnson N;Bradley A;Kotarsky C;Jepng'etich L;Friesner D;Stastny S;Hackney KJ;Nawarathna D
• 通讯作者: Nawarathna D