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个集成诊断系统，旨在快速隔离、 人类临床中循环外泌体 microRNA (miRNA) 生物标志物的检测和多重鉴定 用于早期发现疾病的样本。 miRNA 失调经常在多种疾病中观察到，包括 癌症、心血管疾病和感染。最近的研究还表明 miRNA 可以 通过外泌体从肿瘤细胞分泌到血液中；它们的表达谱可用于对癌症进行分类 类型。因此，循环外泌体 miRNA 被认为是用于早期检测和治疗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产生的技术困难 这些小分子的分析、检测尚未被采用到临床实践中。因此，有必要 开发替代分析策略，与传统方法相比具有更多优势。在这个 项目中，我们提出了一种集成声流控等离子体分子诊断系统，旨在快速 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 新功能可促进疾病的精确临床诊断。该系统集成了两个独特的 技术：（1）声流控技术，可快速高产率、高纯度分离外泌体；和（2） 基于表面增强拉曼散射（SERS）的“逆分子哨兵”（iMS）纳米探针，用于直接 miRNA 检测。虽然与结直肠癌（CRC）相关的 miRNA 将被用作模型系统，但 拟议的项目还将导致普遍适用的护理点诊断技术的开发 对于其他类型的疾病。具体目标是：（1）开发并集成声流系统 外泌体 miRNA 分离； (2) 开发用于外泌体miRNA多重检测的iMS纳米探针 生物标志物； (3) SERS声流系统的技术评估。我们将在合并的基础上再接再厉 我们的跨学科团队的知识建立了准备这种集成所需的技术验证 未来在临床环境中使用的技术。在整个项目中，将开发一个集成系统并 经验证可对临床样本中的多种外泌体 miRNA 进行原位分析，无需 miRNA 提取和扩增。在技​​术验证阶段，结果将与获得的结果进行比较 使用常规检测（例如 qRT-PCR）。该系统将实现无样品制备和“样品到- 答案”分析​​方法基于现有的和经过测试的子组件的集成。这种新的 诊断系统能够加强早期检测和筛查领域的研究和转化 CRC 以外的各种疾病；它非常适合即时检测。拟议的系统 代表了生物医学研究、诊断和筛查领域具有变革潜力的重大创新。