Fingerprinting Cancer via Multiparameter Extracellular Vesicle Analysis

通过多参数细胞外囊泡分析对癌症进行指纹识别

• 批准号: 10325414
• 负责人: Sean German
• 金额: $ 40万
• 依托单位: ELECTRONIC BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10325414
• 关键词: Antibodies; Area; Atomic Force Microscopy; Back; Basic Cancer Research; Biological; Biological Markers; Biological Sciences; Breast Cancer cell line; Cancer Diagnostics; Cell Line; Cells; Cellular Assay; Charge; Collaborations; Collection; Concentration measurement; Consumption; Development; Devices; Diagnostic; Diffuse; Disease; Drug Carriers; Electron Microscopy; Field Flow Fractionation; Fingerprint; Flow Cytometry; Fluorescence; Future; Goals; Health; Heat-Shock Proteins 70; Heterogeneity; Human; Individual; Label; Lasers; Lipids; Liquid substance; MCF7 cell; MDA MB 231; Malignant Neoplasms; Mammalian Cell; Measurement; Measures; Membrane Proteins; Mental disorders; Methods; MicroRNAs; Microscopy; Molecular; Monitor; Neoplasm Metastasis; Nucleic Acids; Oncogenic; Oncology; Parents; Pathogenesis; Pathway interactions; Pattern; Performance; Persons; Phase; Phenotype; Photobleaching; Physiologic pulse; Physiological; Population; Price; Process; Production; Proteins; Research; Research Personnel; Resolution; Role; Sampling; Scanning; Signal Transduction; Small Business Technology Transfer Research; Sorting - Cell Movement; Streptolysins; Surface; System; Techniques; Technology; Time; Transmission Electron Microscopy; Tumor-Derived; United States National Institutes of Health; Universities; Utah; Vesicle; base; biomarker identification; biophysical properties; cancer cell; cryogenics; density; detection platform; distinguished professor; exosome; extracellular vesicles; fluorescence imaging; fluorophore; imaging detection; instrument; intercellular communication; interest; light scattering; liquid biopsy; millisecond; molecular diagnostics; multiplex detection; nanoparticle; nanopore; nanoscale; nervous system disorder; particle; physical property; prevent; professor; programs; prototype; response; statistics; targeted cancer therapy; therapeutic target; tool; tumor progression; tumorigenesis; vesicular release; wasting; zeta potential

Project Summary Electronic BioSciences (EBS) proposes, in collaboration with Distinguished Professor Henry White at the University of Utah, to develop a nanopore-based extracellular vesicle (EV) characterization system capable of high-resolution, single-vesicle, solution-based characterization, sorting, and isolation. EVs are nanometer-scale, cell-derived vesicles that are produced by nearly all mammalian cells under normal physiological conditions and with increased production rates for various disease states (e.g. cancers, neurological diseases, psychiatric disorders, etc.). EVs can be found in most biological fluids and contain proteins, nucleic acids, and lipid molecules that are distinctly dissimilar to the parent cell’s cytoplasmic contents, indicating that exosome loading is not a diffusive or unregulated process. Furthermore, EVs facilitate intercellular communication by transferring these contents into recipient cells to alter the target cells’ phenotype and function. Thus, in addition to their general physiological roles, EVs have important implications in disease pathogenesis, and significant potential as therapeutic targets and drug carriers. Consequently, this has spurred a great deal of interest in new methods to not only characterize the physical properties of EVs (size, zeta potential, surface markers, nucleic acid cargo, and concentration), but increased interest in developing new techniques that have the ability to efficiently and accurately sort/isolate EVs such that the contents/cargo can be profiled in a subpopulation-specific manner. Advances in the areas of EV characterization and sorting/isolation would greatly assist the efforts to understand the general role of EVs in human health, and also benefit efforts aimed at utilizing EVs in molecular diagnostics, biomarker identification, and targeted therapies for cancer. The EV characterization and sorting system that EBS and Professor Henry White will be developing during this program will specifically enable single-particle assessments of an EV sample’s true distribution of size, zeta potential, density of fluorescently tagged surface markers, and nucleic acid cargo, along with the ability to isolate/collect specific EV subpopulations with tightly defined biophysical parameters, all on a single platform. The feasibility of the proposed system will be demonstrated by building a prototype platform and showing its ability to characterize, sort and isolate EV samples. The resulting system will have an ease-of-use, throughput, and price point similar to flow cytometry, dynamic light scattering (DLS), nanoparticle tracking analysis (NTA), or single-pass resistive-pulse techniques, with a resolution comparable to microscopy techniques, e.g. scanning or tunneling electron microscopy (SEM or TEM, respectively) or atomic force microscopy (AFM), yielding a state-of-the-art instrument which will directly enable and advance the field of EV diagnostics.

项目摘要 电子生物科学(EBS)与著名教授亨利·怀特在 开发基于纳米孔的细胞外小泡(EV)表征系统，该系统能够 高分辨率、单泡、基于溶液的表征、分类和分离。电动汽车是纳米级的， 几乎所有哺乳动物细胞在正常生理条件下产生的细胞衍生的囊泡 随着各种疾病(例如癌症、神经疾病、精神疾病)的生产率增加 障碍等)。EVS存在于大多数生物体液中，含有蛋白质、核酸和脂质分子 与亲本细胞的细胞质内容物明显不同，这表明外切体加载不是 扩散的或不受管制的过程。此外，电动汽车通过传输这些信息来促进细胞间的交流 内容物进入受体细胞，改变靶细胞的表型和功能。因此，除了他们的一般 EVS在疾病的发病机制中具有重要的意义，并具有显著的潜力 治疗靶点和药物载体。因此，这激发了人们对新方法的极大兴趣 不仅表征EVS的物理属性(大小、Zeta电位、表面标记、核酸货物、 和专注力)，但对开发新技术的兴趣越来越大，这些新技术能够高效和 准确地分类/隔离电动汽车，以便可以按特定人群的方式对内容物/货物进行分析。 在EV特征和分类/隔离领域的进展将极大地有助于努力了解 电动汽车在人类健康中的一般作用，也有利于旨在利用电动汽车进行分子诊断的努力， 生物标记物的识别和癌症的靶向治疗。EBS电动汽车表征与分类系统 亨利·怀特教授将在这个项目中专门实现单粒子 评估EV样本的大小、Zeta电位、荧光标记表面密度的真实分布 标记和核酸载体，以及分离/收集特定EV亚群的能力 定义的生物物理参数，所有这些都在一个平台上。拟议系统的可行性将是 通过构建原型平台并展示其表征、分类和隔离电动汽车的能力进行了演示 样本。由此产生的系统将具有类似于流式细胞术的易用性、吞吐量和价格点， 动态光散射(DLS)、纳米颗粒跟踪分析(NTA)或单程电阻脉冲技术， 分辨率可与显微镜技术相媲美，例如扫描或隧道电子显微镜(SEM或 或原子力显微镜(AFM)，产生了一种最先进的仪器，它将直接 启用并推进电动汽车诊断领域。