Two-Dimensional Multi-Stage Isotachophoretic Technology for Multiplex Analysis of Cancer Exosomes and Proteins Marker Panel

用于癌症外泌体和蛋白质标记物组多重分析的二维多级等速电泳技术

基本信息

批准号：
10322022
负责人：
WEN-JI DONG
金额：
$ 16.88万
依托单位：
WASHINGTON STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10322022
关键词：
3D Print Address Adopted Antibodies Antigens Area Behavior Binding Biological Blood Blood Circulation CD44 gene Cancer Patient Cancer Prognosis Cancerous Cations Cell secretion Cells Cellular Phone Clinical Communication Consumption Death Rate Detection Devices Diagnosis Disease Distal Early Diagnosis Epidermal Growth Factor Receptor Exhibits Hematopoietic Neoplasms Immune Immune Targeting Lead Liquid substance Malignant Neoplasms Membrane Methods MicroRNAs Microfluidics Modeling Neoplasm Metastasis Normal Cell Paper Pathologic Processes Patients Plasma Proteins Play Point of Care Technology Population Primary Health Care Process Prognosis Proteins Quality of life Research Role Sampling Scheme Screening for cancer Sensitivity and Specificity Serum Serum Proteins Signaling Molecule Survival Rate Techniques Technology Testing Time Tissues Translating Translational Research Tumor Markers Tumor-Derived base biomarker panel cancer biomarkers cancer cell cancer diagnosis cell type clinical diagnosis clinical prognosis clinically significant cost cost effective design early screening exosome extracellular extracellular vesicles improved interdisciplinary approach liquid biopsy malignant breast neoplasm miniaturize multiplex assay neoplastic cell noninvasive diagnosis novel novel strategies point of care protein biomarkers rapid technique success survivin tool translational potential tumor tumor growth tumor progression two-dimensional

项目摘要

Abstract Exosomes are small-sized (30–120 nm) extracellular vesicles. They are secreted by most cell types and play important roles in extracellular communication in normal and pathological processes. The exosomes derived from cancers shuttle signaling molecules (e.g. proteins and miRNAs) from parental cancer cells and tissue to distal recipient cells to reprogram the recipient cells and promote tumor growth and metastasis. Therefore, circulating tumor-derived exosomes carrying signature protein markers of the tumor hold great potential as invaluable liquid biopsy tools for the noninvasive diagnosis of early-stage cancers. Despite their potential clinical significance, translating disease-derived exosomes into point-of-care (POC) applications for the early diagnosis of cancers is hampered by a critical technical barrier: lack of a cost-effective POC approach capable of the simultaneous analysis of specific exosomes and their content markers in clinical samples. Although a number of methods for exosome isolation and characterization have been developed, either they are singly functionalized, nonspecific, laborious, or time-consuming, or they lack the robustness to be adopted as a cost-effective POC technique. Therefore, there is an urgent need for an effective, precise, easy to use, low-cost approach to multiplex POC sample analysis to detect trace levels of specific populations of exosomes released by specific cancer cells at an early stage and comprehensively profile the cancer markers carried by the exosomes. This application aims to fill the gap by taking a multidisciplinary approach to developing a novel, disposable two- dimensional paper-based multistage isotachophoresis (ITP) technology platform capable of the simultaneous analysis of specific target exosomes and exosomal proteins in a cost-effective way. Our objective is to develop an integrated paper-based isotachophoretic platform on which: 1) anionic cascade ITP is used to deplete high- abundance plasma proteins and enrich target exosomes before their capture and analysis; 2) a second ITP process simultaneously analyzes multiplex exosomal proteins released by lysing the exosomes captured in the first ITP process; and 3) a miniaturized smartphone-based detection module quantifies the target exosomes and protein markers captured by novel graded-binding test lines. We expect that integrating effective isolation/identification of specific exosomes with multiplex analysis of their contents in a cost-effective modular platform will provide a robust POC approach to advancing basic and clinical translational research on disease- derived exosomes. Exosomes derived from breast cancer will be used as model targets to validate our technology. The success of this project will not only provide a clinically compatible POC tool for tracking specific exosomes and markers for early screening for cancers, but also prompt research on the profile analysis of exosomal markers for the precision diagnosis of other diseases. Therefore, the technique may have broad translational potential in managing patients with malicious diseases and improving their quality of life.

抽象的外泌体是小型（30-120 nm）细胞外蔬菜。它们被大多数细胞类型分泌在正常和病理过程中细胞外交流中的重要作用。外泌体得出从癌症班车信号分子（例如蛋白质和miRNA）从亲本癌细胞和组织到远端受体细胞重新编程受体细胞并促进肿瘤的生长和转移。所以，携带肿瘤标志物的循环肿瘤衍生的外泌体具有很大的潜力无创的液体活检工具，用于早期癌症的无创诊断。尽管有潜在的临床意义，将疾病衍生的外泌体转化为预防点（POC）的早期诊断癌症的癌症受到关键技术障碍的阻碍：缺乏具有成本效益的POC方法在临床样本中对特定外泌体及其含量标记的同时分析。虽然有很多已经开发了用于外泌体隔离和表征的方法，要么单一功能化，要么非特异性，实验室或时间耗时，或者它们缺乏被用作具有成本效益的POC的鲁棒性技术。因此，迫切需要采取有效，精确，易于使用的低成本方法多重POC样品分析以检测特定于特定的外泌体的特定外泌体的痕量水平癌细胞在早期和综合概况上由外泌体携带的癌症标志物。这应用程序旨在通过采用多学科方法来填补空白基于尺寸的纸张多阶段等齿（ITP）技术平台，能够同时使用以具有成本效益的方式分析特定靶外泌体和外泌体蛋白质。我们的目标是发展基于纸张的集成式等法平台：1）使用阴离子级联iTP来复制高级抽象等离子体蛋白在捕获和分析之前富集靶外泌体； 2）第二个ITP 过程简单地分析了通过在捕获的外泌体释放的多重外泌体蛋白第一个ITP过程； 3）基于智能手机的小型检测模块量化了目标外泌体和新型分级结合测试线捕获的蛋白质标记物。我们期望整合有效在具有成本效益的模块化中对其内容分析的特定外泌体的隔离/鉴定平台将提供强大的POC方法来推进有关疾病的基本和临床转化研究 - 派生的外泌体。源自乳腺癌的外泌体将用作验证我们的模型靶标技术。该项目的成功不仅将提供临床上兼容的POC工具，用于跟踪特定用于癌症早期筛查的外泌体和标记其他疾病精确诊断的外泌体标记。因此，该技术可能具有广泛转化潜力在管理恶意疾病并改善其生活质量的患者方面。