Development and validation of nanoparticle-mediated microfluidic profiling approach for rare cell analysis

项目概要： 循环肿瘤细胞（CTC）从原发性肿瘤脱落到血管系统中，并且已被证明可以 有助于模型系统中转移性病变的形成。因此，监测这些循环细胞 原则上提供了一种真实的实时监测肿瘤转移潜力的方法。类似于 由于个体肿瘤内细胞亚群的异质性，个体内的CTC也表现出 异质性，包含与转移性疾病的发展具有不同相关性的亚群。 最近的研究表明，CTC的特定亚群具有转移潜力，而其他亚群具有转移潜力。 循环上皮细胞的亚群可能是相对良性的。 类似地，CTC上的表面蛋白水平是异质的和动态的：观察到它们与CTC表面蛋白的水平相关。 作为疾病阶段和对治疗反应的函数的变化。特别是，上皮间充质 在CTC中，EMT似乎是一个动态过程，对应于这两个EMT的标记物 状态不同，并导致CTC的表型异质性。使用微流体装置， 谷（VV）芯片，这是在我们的小组开发的，我们现在有能力分析CTC人口从 血液样品，并基于表面标志物的表达分选这些细胞。这项新技术 使我们能够捕获和研究各种EMT范围内的CTC。 在本提案中，我们的目标是将VV芯片技术全面发展成为CTC的完全集成器件 群体分析、CTC检测和分子分析。这将通过整合 配套技术允许对CTCs进行灵敏的芯片上电化学检测和基因分析。 将研究器械的生产方法，以进行大规模生产。此外，本发明还提供了一种方法， 将开发样品分析和检测的自动化，使该设备能够在一个 临床或研究环境。最后，该设备将通过前列腺和乳腺的临床样本进行验证 癌症患者。 该项目将包括一个由六名研究人员和临床医生组成的多学科团队的合作， 开发、生产和临床试验。作为一个团队，研究人员将致力于开发和验证 这个诊断平台。在该项目完成后，将生产一种临床研究工具， 对患者的CTC群体进行分析，并为此提供关于EMT的分子和遗传信息。 在具有捕获、分析和检测功能的单个自动化设备中填充。

Project Summary: Circulating tumour cells (CTC) are shed into the vasculature from primary tumours, and have been shown to contribute to the formation of metastatic lesions in model systems. Monitoring these circulating cells therefore presents, in principle, a means to monitor a tumour's metastatic potential in real time. Similar to the heterogeneity of cellular subpopulations within an individual tumour, CTCs within an individual also exhibit heterogeneity, containing subpopulations having varying relevance to the development of metastatic disease. Recent studies show that specific subpopulations of CTCs possess metastatic potential, while other subpopulations of circulating epithelial cells may be relatively benign. Similarly, the levels of surface proteins on CTCs are heterogeneous and dynamic: they are observed to change as a function of disease stage and response to therapy. In particular, the epithelial-mesenchymal transition (EMT) appears to be a dynamic process in CTCs, and the markers that correspond to these two states vary and contribute to the phenotypic heterogeneity of CTCs. Using a microfluidic device, the velocity valley (VV) chip, that was developed in our group, we now have the ability to profile a CTC population from blood samples and by sorting these cells based on expression of surface markers. This novel technology has enabled us to capture and study CTCs within various ranges of EMT. In this proposal, our goal is to fully develop the VV chip technology into a fully integrated device for CTC population profiling, CTC detection, and molecular analysis. This will be accomplished through integration of companion technologies allowing for sensitive on-chip electrochemical detection and genetic analysis of CTCs. Manufacturing methods for the device will be investigated for production at high-scale. In addition, automation for sample analysis and detection will be developed enabling the full realization of the device in a clinical or research setting. Finally the device will be validated with clinical samples from prostate and breast cancer patients. This project will include the collaboration of a multidisciplinary team of six researchers and clinicians for device development, manufacturing, and clinical testing. As a team, the researchers will work to develop and validate this diagnostic platform. At the completion of this project a clinical research tool will be produced capable of profiling a patient's CTC population and providing molecular and genetic information on the EMT for that population in a single automated device with capture, profiling, and detection capabilities.