Integrated Acoustofluidic Plasmonic Molecular Diagnostic System for Detecting MicroRNA Biomarkers

用于检测 MicroRNA 生物标志物的集成声流控等离子体分子诊断系统

• 批准号: 10542811
• 负责人: Tuan Vo-Dinh
• 金额: $ 42.02万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10542811
• 关键词: Acoustics; Adopted; Area; Benchmarking; Biological; Biological Assay; Biological Markers; Biological Models; Biomedical Research; Blood Circulation; Blood specimen; Body Fluids; Cancer Diagnostics; Cardiovascular Diseases; Cardiovascular system; Cell secretion; Centrifugation; Characteristics; Chylomicrons; Clinical; Colorectal Cancer; Communicable Diseases; Consumption; Detection; Development; Devices; Diagnostic; Disease; Early Diagnosis; Early treatment; Encapsulated; Evaluation; Future; Gastrointestinal tract structure; Gene Expression; Goals; Human; Human Resources; In Situ; Infection; Investigation; Knowledge; Lipoprotein (a); Lipoproteins; Liquid substance; Low-Density Lipoproteins; Malignant Neoplasms; Measures; Metaplasia; Methods; MicroRNAs; Microfluidics; Molecular; Molecular Analysis; Molecular Biology; Monitor; Neurodegenerative Disorders; Northern Blotting; Output; Pathology; Performance; Phase; Preparation; Procedures; Process; Proteins; Quantitative Reverse Transcriptase PCR; Research; Research Personnel; Sampling; Sentinel; Sorting; Specificity; Specimen; Speed; Statistical Data Interpretation; Surface; Symptoms; System; Systems Analysis; Systems Integration; Techniques; Technology; Testing; Time; Training; Translations; Tumor-Derived; Ultracentrifugation; Untranslated RNA; Validation; Vesicle; Whole Blood; accurate diagnosis; biobank; biological research; biomaterial compatibility; cancer classification; cancer diagnosis; cancer type; circulating microRNA; clinical application; clinical diagnosis; clinical practice; clinical translation; clinically relevant; colon cancer patients; cost effective; density; design; detection platform; diagnostic platform; diagnostic technologies; disease classification; disease diagnosis; disease diagnostic; early detection biomarkers; early screening; exosome; improved; innovation; interactive feedback; interdisciplinary collaboration; microRNA biomarkers; molecular diagnostics; molecular marker; mortality; multiplex detection; nanobiosensor; nanoprobe; nanoscale; neoplastic cell; new technology; novel; novel diagnostics; performance tests; physical property; plasmonics; point of care; point of care testing; point-of-care diagnostics; posttranscriptional; rapid diagnosis; screening; small molecule; targeted biomarker

The goal of this project is to develop an integrated diagnostic system designed for rapid isolation, detection, and multiplexed identification of circulating exosomal microRNA (miRNA) biomarkers in human clinical samples for early detection of diseases. Dysregulation of miRNAs is often observed in various diseases including cancer, cardiovascular illnesses, and infection. Recent studies have also demonstrated that miRNAs can be secreted from tumor cells into bloodstream via exosomes; their expression profiles can be used to classify cancer types. Therefore, circulating exosomal miRNAs are considered promising biomarkers for early detection and diagnosis of cancer. However, because of technical difficulties arising from exosome isolation and miRNA analysis, detection of these small molecules has not been adopted into clinical practice. Thus, there is a need to develop alternative analysis strategies that could offer more advantages over conventional methods. In this project, we propose an integrated acoustofluidic plasmonic molecular diagnostic system designed for rapid isolation and multiplexed detection of exosomal miRNA biomarkers. The cross-disciplinary approach will provide new capabilities to advance the precise clinical diagnosis of diseases. This system integrates two unique technologies: (1) acoustofluidic technology that can rapidly isolate exosomes with high yield and purity; and (2) surface-enhanced Raman scattering (SERS)-based “Inverse Molecular Sentinel” (iMS) nanoprobes for direct miRNA detection. Although miRNAs related to colorectal cancer (CRC) will be used as the model system, the proposed project will also lead to the development of a generally applicable point-of-care diagnostic technology for other types of diseases. The specific aims are: (1) Develop and integrate an acoustofluidic system for exosomal miRNA isolation; (2) Develop iMS nanoprobes for multiplexed detection of exosomal miRNA biomarkers; and (3) Technical Evaluation of the SERS-acoustofluidic system. We will build upon the combined knowledge of our interdisciplinary team establishing the technical validation required to ready this integrated technology for future use in clinical settings. Throughout this project, an integrated system will be developed and validated for in-situ analysis of multiple exosomal miRNAs from clinical samples without the need for miRNA extraction and amplification. In the technical validation phase, the results will be compared to those obtained using conventional assays (e.g. qRT-PCR). The system will lead to a no-sample preparation and “sample-to- answer” analysis approach that is based on the integration of existing and tested sub-components. This new diagnostic system is capable of enhancing research and translation in the areas of early detection and screening of various diseases beyond CRC; it is exceedingly well suited for point-of-care testing. The proposed system represents a major innovation with transformative potential in biomedical research, diagnostics, and screening.

该项目的目标是开发一种用于快速隔离的综合诊断系统， 人类临床中循环外泌体微小RNA（miRNA）生物标志物的检测和多重鉴定 用于早期检测疾病的样本。在多种疾病中经常观察到miRNAs的失调，包括 癌症心血管疾病和感染最近的研究也表明，miRNA可以被 通过外泌体从肿瘤细胞分泌到血流中;它们的表达谱可用于癌症分类 类型因此，循环外泌体miRNA被认为是用于早期检测的有希望的生物标志物， 癌症的诊断。然而，由于外泌体分离和miRNA的技术困难， 分析，检测这些小分子还没有采用到临床实践。因此，有必要 开发可提供比传统方法更多优势的替代分析策略。在这 项目，我们提出了一个集成的声流等离子体分子诊断系统，设计用于快速 外泌体miRNA生物标志物的分离和多重检测。跨学科方法将提供 提高疾病精确临床诊断的新能力。该系统集成了两个独特的 技术：（1）声流体技术，可以快速分离外泌体，产量和纯度高;和（2） 基于表面增强拉曼散射（Sers）的“反分子哨兵”（iMS）纳米探针，用于直接 miRNA检测。尽管与结直肠癌（CRC）相关的miRNA将被用作模型系统，但是， 拟议的项目还将导致普遍适用的即时诊断技术的发展 其他类型的疾病。具体目标是：（1）开发和集成声流控系统， （2）开发iMS纳米探针用于外泌体miRNA的多重检测 生物标志物;和（3）SERS声流体系统的技术评估。我们将建立在 我们的跨学科团队的知识，建立所需的技术验证，准备这个集成 未来在临床环境中使用的技术。在整个项目中，将开发一个综合系统， 经验证可用于临床样品中多种外泌体miRNA的原位分析，而无需miRNA 提取和扩增。在技术验证阶段，将把结果与所获得的结果进行比较 使用常规测定（例如qRT-PCR）。该系统将导致无样品制备和“样品到样品”， 答案”分析方法是基于现有的和测试的子组件的集成。这个新 诊断系统能够加强早期检测和筛查领域的研究和翻译 除了CRC之外的各种疾病;它非常适合即时检测。所提出的系统 这是一项重大创新，在生物医学研究、诊断和筛查方面具有变革潜力。