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: 10580345
• 负责人: Tuan Vo-Dinh
• 金额: $ 3.44万
• 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 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10580345
• 关键词: Acoustics; Adopted; Area; Benchmarking; Biological Assay; Biological Markers; Biological Models; Biomedical Research; Blood Circulation; Blood specimen; Cancer Diagnostics; Cardiovascular Diseases; Cardiovascular system; Centrifugation; Chylomicrons; Clinical; Colorectal Cancer; Communicable Diseases; Detection; Development; Devices; Disease; Early Diagnosis; Early treatment; Evaluation; Goals; Human; Human Resources; In Situ; Investigation; Knowledge; Lipoprotein (a); Lipoproteins; Low-Density Lipoproteins; Malignant Neoplasms; Measures; Messenger RNA; Methods; MicroRNAs; Molecular; Molecular Analysis; Monitor; Output; Pathology; Performance; Phase; Preparation; Proteins; Quantitative Reverse Transcriptase PCR; Research; Sampling; Sentinel; Specificity; Specimen; Speed; Statistical Data Interpretation; Surface; Symptoms; System; Systems Analysis; Techniques; Technology; Testing; Training; Translations; Validation; Whole Blood; accurate diagnosis; base; biobank; biological research; cancer diagnosis; cancer type; clinical application; clinical diagnosis; clinical practice; clinical translation; clinically relevant; colon cancer patients; cost effective; density; design; diagnostic platform; diagnostic screening; diagnostic technologies; disease diagnostic; early detection biomarkers; early screening; exosome; improved; innovation; interactive feedback; microRNA biomarkers; molecular diagnostics; molecular marker; mortality; multiplex detection; nanoprobe; neoplastic cell; new technology; novel; performance tests; physical property; plasmonics; point of care; point-of-care diagnostics; rapid diagnosis; small molecule

ABSTRACT The goal of this project is to develop an Integrated acoustofluidic plasmonic molecular 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 infectious diseases. For instance, recent studies have also demonstrated that miRNAs can be secreted from tumor cells into bloodstream via exosomes, and 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, these small molecules have not been adopted into clinical practice because of the technical difficulties involved in the isolation of exosomes and the analysis of exosomal miRNAs. Thus, there is a need to develop alternative analyzing strategies that could offer more advantages over conventional methods. In this regard, 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 cancer. This system integrates two unique but proven technologies: (1) the 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 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 and establish the technical validation required to ready this integrated technology for use in clinical settings. Throughout this project, an integrated acoustofluidic plasmonic 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 result will be compared to that obtained using conventional assays (e.g. qRT-PCR). The proposed system will lead to a no-sample preparation and “sample- to-answer” analysis approach that is based on the integration of existing sub-components, many of which have already been separately tested in similar applications. This new integrated molecular diagnostic system is capable of enhancing research and translation in the areas of early detection and screening of various cancers beyond CRC, and is ultimately well suited for point-of-care. The proposed molecular analysis system represents a major innovation that has a transformative potential in biomedical research, diagnostics and screening.

摘要 本计画的目标是发展一套整合式声流电浆分子诊断系统 设计用于快速分离、检测和多重鉴定循环外泌体microRNA（miRNA） 用于早期检测疾病的人类临床样品中的生物标志物。经常观察到miRNAs的失调 癌症、心血管疾病和传染病等多种疾病。例如，最近 研究还表明miRNA可以通过外来体从肿瘤细胞分泌到血流中， 并且它们的表达谱可用于分类癌症类型。因此，循环外泌体miRNA是 被认为是早期检测和诊断癌症的有前途的生物标志物。然而，这些小分子 由于在分离中涉及的技术困难， 外泌体和外泌体miRNA的分析。因此，有必要开发替代分析 可以提供比传统方法更多优势的策略。在这方面，我们提出了一个综合的 声流等离子体分子诊断系统设计用于快速分离和多重检测 外泌体miRNA生物标志物。跨学科方法将提供新的能力， 癌症的精确临床诊断。该系统集成了两种独特但经过验证的技术：（1） 声流体技术，其可以以高产率和纯度快速分离外泌体，和（2）表面增强的 基于拉曼散射（SERS）的"反向分子哨兵"（iMS）纳米探针用于直接miRNA检测。 虽然与结直肠癌（CRC）相关的miRNAs将被用作模型系统，但拟议的项目将 导致开发出普遍适用于其他类型疾病的即时诊断技术。 具体目标是：（1）开发并整合一套用于外泌体miRNA分离的声流控系统; (2)开发iMS纳米探针用于外泌体miRNA生物标志物的多重检测;以及（3）技术 SERS-声流体系统的评估。我们将建立在我们的综合知识， 跨学科的团队，并建立技术验证所需的准备使用这种综合技术 在临床环境中。在整个项目中，将开发一个集成的声流体等离子体系统， 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，将结果与使用 常规测定（例如qRT-PCR）。拟议的系统将导致无样品制备和"样品- 基于现有子组件的集成的"回答"分析方法，其中许多子组件具有 已经在类似的应用中进行了单独测试。这种新的集成分子诊断系统是 能够加强各种癌症的早期检测和筛查领域的研究和翻译 超越CRC，最终非常适合床旁护理。所提出的分子分析系统代表了 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。

项目成果

Smartphone-Based Device for Colorimetric Detection of MicroRNA Biomarkers Using Nanoparticle-Based Assay.

• DOI: 10.3390/s21238044
• 发表时间: 2021-12-01
• 期刊: Sensors (Basel, Switzerland)
• 作者: Krishnan T;Wang HN;Vo-Dinh T
• 通讯作者: Vo-Dinh T

A sound approach to advancing healthcare systems: the future of biomedical acoustics.

• DOI: 10.1038/s41467-022-31014-y
• 发表时间: 2022-06-16
• 期刊: Nature communications
• 影响因子: 16.6

Acoustofluidic Holography for Micro- to Nanoscale Particle Manipulation.

• DOI: 10.1021/acsnano.0c03754
• 发表时间: 2020-11-24
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Gu Y;Chen C;Rufo J;Shen C;Wang Z;Huang PH;Fu H;Zhang P;Cummer SA;Tian Z;Huang TJ
• 通讯作者: Huang TJ

A Cell-Phone-Based Acoustofluidic Platform for Quantitative Point-of-Care Testing.

• DOI: 10.1021/acsnano.9b08349
• 发表时间: 2020-03
• 期刊: ACS nano
• 影响因子: 17.1
• 作者: Liying Zhang;Zhenhua Tian;Hunter Bachman;Peiran Zhang;T. Huang

Acoustofluidics for simultaneous nanoparticle-based drug loading and exosome encapsulation.

• DOI: 10.1038/s41378-022-00374-2
• 发表时间: 2022
• 期刊: MICROSYSTEMS & NANOENGINEERING
• 影响因子: 7.9
• 作者: Wang, Zeyu;Rich, Joseph;Hao, Nanjing;Gu, Yuyang;Chen, Chuyi;Yang, Shujie;Zhang, Peiran;Huang, Tony Jun
• 通讯作者: Huang, Tony Jun