Development of plasmon-enhanced biosensing for multiplexed profiling of extracellular vesicles

用于细胞外囊泡多重分析的等离子体增强生物传感的发展

• 批准号: 10685632
• 负责人: Hyungsoon Im
• 金额: $ 35.7万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10685632
• 关键词: Address; Basic Science; Binding; Biological; Biological Assay; Biological Markers; Biological Models; Biosensing Techniques; Blood; Body Fluids; Cancer cell line; Cardiovascular Diseases; Cardiovascular system; Cells; Cerebrospinal Fluid; Clinical; Clinical Research; Color; Communicable Diseases; Cytolysis; Detection; Development; Disease; Emerging Technologies; Fluorescence; Generations; Genetic Materials; Goals; Heterogeneity; Human; Individual; Integral Membrane Protein; Label; Lipids; Liquid substance; Malignant Neoplasms; Malignant neoplasm of ovary; Membrane; Methods; Modeling; Molecular; Molecular Probes; Monitor; Names; Nerve Degeneration; Neurodegenerative Disorders; Performance; Periodicals; Phospholipids; Plasma; Procedures; Process; Production; Proteins; RNA; RNA marker; Reproducibility; Saliva; Sampling; Signal Transduction; Structure; Surface Plasmon Resonance; System; Technology; Technology Assessment; Testing; Tumor-Derived; Urine; Variant; Vesicle; cancer biomarkers; cancer diagnosis; cell type; circulating biomarkers; clinical application; clinically relevant; design; detection assay; detection platform; detection sensitivity; exosome; experimental study; extracellular vesicles; improved; innovation; insight; liquid biopsy; minimally invasive; molecular diagnostics; molecular marker; nano; nanoGold; nanoplasmonic; nanopore; nanoscale; new technology; next generation; noninvasive diagnosis; pre-clinical; protein biomarkers; prototype; sensor; sensor technology; tool; transmission process; treatment response; tumor

Extracellular vesicles (EVs) present new opportunities for molecular diagnostics from non-invasive liquid biopsies. These cell-derived membrane-bound vesicles are abundantly present in biological fluids. EVs carry cell-specific cargos (e.g., lipids, proteins, and genetic materials), which can be harnessed to probe the molecular status of their cellular origins. EV analyses, however, pose unique technical challenges due to EVs' nanometer-sizes and presence in a vast biological background. EV analyses, however, pose unique technical challenges due to EVs' nanometer-sizes and presence in a vast biological background. While new technologies for EV analysis have been developed, fundamental limitations still remain, including i) low sensitivity limited to bulk analyses; ii) necessities of EV lysis for detecting markers inside of EVs; iii) lack of multiplexed analysis on protein and RNA markers; and iv) a separate EV isolation process required prior to the assay. The overall goal of this application is to overcome these technical challenges and develop a new platform that enables multiplexed analyses of EV protein and RNA markers in individual EVs. We previously developed a nanoplasmonic EV sensing platform based on transmission surface plasmon resonance through periodic nanohole gratings. We showed that the nanoplasmonic sensors could rapidly and sensitively detect disease-specific EVs directly from clinical samples. In this project, we will further advance the technology for robust multiplexed EV analysis and implement on-chip EV isolation to achieve simple assay procedures and good reproducibility. We will validate the system using well-established preclinical and clinical samples to demonstrate the feasibility and potential of the new technology for clinical applications. Successful completion of the project will produce a highly sensitive sensing platform for multiplexed EV analysis. The development of such a technology could offer additional insight into understanding subtypes, heterogeneity, and production dynamics of EVs during disease development and progression. The gained insights will pave the way for expanding EV studies to various diseases, further broadening the scope of EV applications in clinical settings.

细胞外囊泡（EV）为非侵入性液体提供了分子诊断的新机会 活检。这些细胞衍生的膜结合的囊泡在生物流体中大量存在。电动汽车携带 细胞特异性的嘉戈斯（例如脂质，蛋白质和遗传材料），可以利用以探测 其细胞起源的分子状态。但是，EV分析由于电动汽车而构成独特的技术挑战 纳米尺寸和在广阔的生物学背景下的存在。 EV分析构成独特的技术 由于电动汽车的纳米尺寸和在巨大的生物学背景下的存在引起的挑战。虽然新 已经开发了电动汽车分析的技术，仍然存在基本限制，包括i）低 灵敏度仅限于批量分析； ii）EV溶解的必要性用于检测电动汽车内部的标记； iii）缺乏 对蛋白质和RNA标记的多重分析； iv）在此之前需要一个单独的EV隔离过程 测定。该应用程序的总体目标是克服这些技术挑战并发展新的 可以在单个EV中对EV蛋白和RNA标记的多路复用分析的平台。我们以前 基于传输表面等离子体共振，开发了一个纳米质感应传感平台 周期性的纳米光栅。我们表明纳米质传感器可以快速而灵敏地检测 直接来自临床样本的疾病特异性电动汽车。在这个项目中，我们将进一步推进技术 强大的多路复用电动汽车分析和实施片上电动汽车隔离，以实现简单的测定程序和 良好的可重复性。我们将使用公认的临床前和临床样本验证系统 证明新技术在临床应用中的可行性和潜力。成功完成 该项目将产生一个高度敏感的传感平台，用于多路复用EV分析。发展的发展 这样的技术可以为了解亚型，异质性和生产提供更多的见解 疾病发育和进展过程中电动汽车的动力学。获得的见解将为 将电动汽车研究扩展到各种疾病，进一步扩大了临床环境中电动汽车应用的范围。