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）为非侵入性液体分子诊断提供了新的机会 活检。这些细胞源性膜结合囊泡大量存在于生物体液中。电动汽车携带 细胞特异性货物（例如脂质、蛋白质和遗传物质），可用于探测 它们的细胞起源的分子状态。然而，由于电动汽车的 纳米尺寸和存在于广阔的生物背景中。然而，EV 分析提出了独特的技术 由于电动汽车的纳米尺寸和存在于广阔的生物背景中，电动汽车面临着挑战。虽然是新的 EV 分析技术已经开发出来，但基本的局限性仍然存在，包括 i) 低 灵敏度仅限于批量分析； ii) EV 裂解检测 EV 内部标记的必要性； iii) 缺乏 蛋白质和 RNA 标记物的多重分析； iv) 在执行之前需要一个单独的 EV 隔离过程 化验。该应用程序的总体目标是克服这些技术挑战并开发一种新的 该平台能够对单个 EV 中的 EV 蛋白和 RNA 标记物进行多重分析。我们之前 开发了基于传输表面等离子体共振的纳米等离子体EV传感平台 周期性纳米孔光栅。我们证明纳米等离子体传感器可以快速、灵敏地检测 直接来自临床样本的疾病特异性 EV。在这个项目中，我们将进一步推进技术 强大的多重 EV 分析并实现片上 EV 隔离，以实现简单的检测程序和 良好的重现性。我们将使用成熟的临床前和临床样本来验证该系统 展示新技术临床应用的可行性和潜力。顺利完成 该项目将生产一个用于多重电动汽车分析的高灵敏度传感平台。的发展 这种技术可以为理解亚型、异质性和生产提供更多见解 EV 在疾病发生和进展过程中的动态。所获得的见解将为 将EV研究扩展到各种疾病，进一步扩大EV在临床中的应用范围。