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; 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.

该项目的目的是开发一个旨在快速隔离的综合诊断系统， 人类临床中循环外泌体microRNA（miRNA）生物标志物的检测和多路复用鉴定 样品以早期发现疾病。在包括 癌症，心血管疾病和感染。最近的研究还表明，miRNA可以是 通过外泌体从肿瘤细胞分泌到血液中；它们的表达曲线可用于对癌症进行分类 类型。因此，循环外泌体miRNA被认为是有希望的早期检测生物标志物和 诊断癌症。但是，由于外部隔离和miRNA引起的技术困难 分析，这些小分子的检测尚未用于临床实践。那是有必要的 制定替代分析策略，这些策略可以比传统方法更具优势。在这个 项目，我们提出了一个综合的声流体塑料分子诊断系统，旨在快速 外泌体miRNA生物标志物的分离和多路复用检测。跨学科的方法将提供 推进疾病精确临床诊断的新功能。该系统集成了两个独特的 技术：（1）可以快速分离出高产量和纯度的外泌体的声流体技术； （2） 基于表面增强的拉曼散射（SERS）基于直接的“逆分子前哨”（IMS）纳米探针 miRNA检测。尽管与结直肠癌（CRC）相关的miRNA将用作模型系统，但 拟议的项目还将导致开发普遍适用的护理点诊断技术 对于其他类型的疾病。具体目的是：（1）开发并整合了用于 外泌体miRNA分离； （2）开发IMS纳米探针用于多路复用的外泌体miRNA 生物标志物； （3）SERS-ACOUSTOFLUIDIC系统的技术评估。我们将建立在总和的基础上 了解我们的跨学科团队，建立了准备综合性所需的技术验证 用于临床环境中未来使用的技术。通过该项目，将开发一个集成系统，并 经过验证，用于对临床样品的多个外泌体miRNA的原位分析，而无需miRNA 提取和放大。在技​​术验证阶段，将将结果与获得的结果进行比较 使用常规测定（例如QRT-PCR）。该系统将导致无样本制备和“样本到样本 答案“分析方法基于现有和测试的子组件的整合。 诊断系统能够增强早期检测和筛查领域的研究和翻译 CRC以外的各种疾病；它非常适合护理点测试。提出的系统 代表了生物医学研究，诊断和筛查中具有变革潜力的主要创新。