Multiplexed Profiling of Single-Cell Secreted Exosomes

单细胞分泌的外泌体的多重分析

• 批准号: 9360660
• 负责人: Yong Zeng
• 金额: $ 18.94万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 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/9360660
• 关键词: Address; Behavior; Biogenesis; Biological Assay; Biological Markers; Biological Process; Biology; Cancer cell line; Cell Communication; Cell Culture Techniques; Cell physiology; Cell secretion; Cells; Characteristics; Classification; Clinical; Development; Disease Pathway; Drug resistance; Early Diagnosis; Eukaryotic Cell; Exhibits; Goals; Gold; Heterogeneity; Human; Immune response; Lab-On-A-Chips; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of ovary; Mammalian Cell; Masks; Measurement; Measures; Mediating; Membrane; Methods; MicroRNAs; Microfluidics; Mission; Modification; Molecular; Molecular Analysis; Molecular Profiling; Monitor; Nanotechnology; Neoplasm Metastasis; Outcome; Pathologic; Pathway interactions; Phenotype; Play; Population; Precision therapeutics; Property; Proteins; Reproducibility; Research; Resistance development; Role; Signal Pathway; Signal Transduction; System; Systems Analysis; Technology; Time; Translating; Tumor-Derived; United States National Institutes of Health; Vesicle; Work; analog; base; cancer cell; cancer diagnosis; digital; drug development; effusion; exosome; expectation; high throughput analysis; innovation; intercellular communication; liquid biopsy; neoplastic cell; protein profiling; single cell analysis; stem; success; tool; trait; tumor; tumor growth; tumor heterogeneity; tumor microenvironment; tumorigenesis

SUMMARY Most eukaryotic cells secrete numerous membrane-derived vesicles of 30-150 nm in size termed exosomes. As an emerging mechanism for cell-to-cell communication, exosomes have been recently found to play important roles in a wide range of biological processes, including cancer development and metastasis. For instance, increasing evidences support the cancer-derived exosomes can reprogram the behavior of recipient cells to promote tumor growth and metastasis. Despite the significance of exosomes, our understanding of their biogenesis, molecular classification, and biological functions remain very limited. One of the challenges is to analyze exosomes released from single cells. Because cells in a tumor are known to be remarkably heterogeneous, single-cell analysis of exosomes is crucial to understanding their pathological roles in cancer. However, current “gold standard” methods can only perform ensemble measurements of exosomes released from a large cell population because of their poor isolation yield, insufficient analysis sensitivity and low throughput. In this proposal, the PI aims to develop for the first time a high-throughput single cell exosome analysis system (SCEAS) capable of probing the secretion and molecular composition of exosomes at the single cell level. The goal will be achieved via two specific aims: 1) Develop a microfluidic digital barcode system for multiplexed, ultrasensitive exosome profiling; and 2) Establish a Single Cell Exosome Analysis System (SCEAS) for quantitative profiling of exosomes derived from single cancer cells. Success of the work will yield a key tool to enable the studies of heterogeneous exosome release by tumor cells at the single cell level, which would facilitate better understanding of intercellular signaling pathways underlying cancer development, metastasis, and drug resistance.

总结 大多数真核细胞分泌许多大小为30-150 nm的膜衍生囊泡，称为外泌体。 作为一种新兴的细胞间通讯机制，外泌体最近被发现在细胞间通讯中发挥重要作用。 在包括癌症发展和转移在内的广泛的生物过程中起重要作用。为 例如，越来越多的证据支持癌症来源的外泌体可以重新编程受体的行为， 细胞促进肿瘤生长和转移。尽管外泌体的重要性，我们的理解， 它们的生物起源、分子分类和生物学功能仍然非常有限。挑战之一是 来分析单细胞释放的外泌体因为肿瘤中的细胞 外来体的异质性单细胞分析对于理解它们在癌症中的病理作用至关重要。 然而，目前的“金标准”方法只能对释放的外来体进行系综测量。 因为它们的分离产率差、分析灵敏度不足和低的 吞吐量在这项提案中，PI的目标是首次开发一种高通量的单细胞外泌体， 该分析系统（SCEAS）能够探测外泌体的分泌和分子组成， 单细胞水平。该目标将通过两个具体目标实现：1）开发微流体数字条形码 用于多重、超灵敏外泌体分析的系统;和2）建立单细胞外泌体分析 系统（SCEAS）用于定量分析来源于单个癌细胞的外泌体。工作成功 将产生一个关键的工具，使研究异质外泌体释放的肿瘤细胞在单细胞 水平，这将有助于更好地了解潜在的癌症细胞间信号通路 发展、转移和耐药性。