Characterization of Biological Nanoparticle Subsets

生物纳米颗粒子集的表征

• 批准号: 10014708
• 负责人: Jennifer Jones
• 金额: $ 152.27万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10014708
• 关键词: B-Lymphocytes; Biological; Biological Markers; Biology; Blood; Calibration; Cells; Clinical; Data; Detection; Dimensions; Epitopes; FOLH1 gene; Flow Cytometry; Fluorescence; Goals; Hypoxia; Immune; Immune response; Immunobiology; Immunotherapy; Individual; Label; Laboratories; Leukocytes; Malignant Neoplasms; Methods; MicroRNAs; Molecular; Monitor; Monoclonal Antibodies; Plasma; Population; Proteins; Protocols documentation; RNA; Radiation; Reporting; Research; Resolution; Robotics; Sorting - Cell Movement; Source; Surveys; T-Lymphocyte; TACSTD1 gene; TGFB1 gene; Technology; Tumor Biology; Tumor stage; Vaccines; Vesicle; analysis pipeline; base; burden of illness; cancer therapy; exosome; extracellular vesicles; high dimensionality; improved; instrumentation; nanoparticle; neoplastic cell; novel marker; particle; programs; receptor; response; single molecule; stressor; therapeutic target; treatment response; tumor; tumor progression

During the past year, we have developed a comprehensive pipeline for EV analysis, which we refer to as the Multiplex-to-Single-EV-Analysis (Mt-SEA) pipeline. This Mt-SEA pipeline includes multiplex high dimensional EV surveys as well as high resolution single EV studies, including nanoFACS protocols for fluorescence detection and calibration, for counting, and for high-throughput sorting, to sort EV subsets from plasma and other biofluids by nanoFACS. Using multiple distinct vesicle populations, we find that nanoFACS sorting is uniquely able to produce high fidelity EV subsets (95-97% pure), with functional biological cargo (RNA and protein). Our current focus is exclusively on application of this method for clinically important studies, for the study of cancer disease burden, metastatic potential, and responses to treatment. Furthermore, we have devised a new class of labels (Molecular NanoTags) that can be used with nanoFACS, not only to improve the detection of EV epitopes, but also to enable single molecule counting by flow cytometry. The specialized instrumentation, expertise, and biospecimen pipeline that we have assembled and are continuing to improve for this nanoFACS program is a unique and distinctive asset for NCI.

在过去的一年里，我们已经开发了一个全面的EV分析管道，我们称之为多路到单EV分析（Mt-SEA）管道。这个Mt-SEA管道包括多重高维EV调查以及高分辨率单个EV研究，包括用于荧光检测和校准、计数和高通量分选的纳米facs协议，通过纳米facs从血浆和其他生物流体中分选EV亚群。使用多个不同的囊泡群体，我们发现nanoFACS分选能够独特地产生高保真的EV亚群（纯度为95-97%），并具有功能性生物货物（RNA和蛋白质）。我们目前的重点是将这种方法应用于临床重要研究，研究癌症疾病负担、转移潜力和对治疗的反应。此外，我们设计了一类新的标签（分子纳米标签），可以与纳米facs一起使用，不仅可以提高EV表位的检测，而且可以通过流式细胞术进行单分子计数。专门的仪器、专业知识和生物标本管道，我们已经组装并正在继续改进这个纳米facs计划，是NCI独特而独特的资产。