SORTING AND PROTEOMIC CHARACTERIZATION OF CIRCULATING TUMOR CELLS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 原发肿瘤的细胞以远处器官转移的形式扩散，最终导致了90%的与癌症有关的死亡，仅在美国每年就有50多万人死亡。因此，转移是癌症研究领域需要解决的最重要的生物学问题之一。虽然原发恶性肿瘤和外周转移之间的细胞联系已经以外周血中循环肿瘤细胞(CTCs)的形式建立起来，但在破译这一非常独特的细胞群体的生物学方面仍然是一个重大挑战，并最终提高我们系统水平对“癌症如何扩散和死亡”的理解。鉴于超过85%的主要癌症，包括肺癌，起源于其上皮细胞，我们将关注转移性肺癌患者外周血中存在的上皮性CTCs。我们最近展示了一种新的微流控细胞分选技术，可以有效地分离全血中频率低至109的CTCs，细胞询问速度为每秒1到1000万个细胞。在这项提案中，我们计划为微流控分选设备增加新的功能，并针对两个基础和临床研究应用。具体地说，我们计划研究肺癌不同阶段转移患者分离的CTCs的蛋白质组特征，并优化从从外周血分离的少数CTC中鉴定原发肿瘤表型的分离和分析技术，最终目标是将微流控和蛋白质组学技术引入临床癌症药物。 我们计划使用一种将尖端技术和生物学相结合的方法来解决从癌症患者分离的CTC的分离、纯化和蛋白质组样本制备问题。这个合作项目的具体目标是设计、制造和表征一种微流控芯片，用于从捕获的CTC开始准备用于蛋白质组分析的样本，并根据蛋白质组特征识别CTC的特征表型。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Dissemination of cells from the primary tumor in the form of metastasis in distant organs is ultimately responsible for 90% of cancer-related deaths, representing over 500,000 people dying every year in the United States alone. Consequently, metastases are among the most important biological problems to address in the area of cancer research. Although a cellular link between the primary malignant tumor and the peripheral metastases has been established in the form of circulating tumor cells (CTCs) in peripheral blood, a major challenge remains in deciphering the biology of this very unique cellular population, and ultimately, improving our systems level understanding of "how cancer spreads and kills." Given that more than 85% of major cancers, including lung cancer, originate from their epithelial cells, our focus will be the epithelial CTCs present in peripheral blood of the patients with metastatic lung cancer. We have recently demonstrated a new microfluidic cell sorting technology for efficient isolation of CTCs having frequencies as low as 1 in 109 in whole blood, at a cell interrogation rate of 1 to 10 million cells per second. In this proposal we plan to add new features to the microfluidic sorting device and target two basic and clinical research applications. Specifically, we plan to study the proteomic signature of the separated CTCs in lung cancer patients with metastatic disease at different stages, and optimize the separation and analysis technology for the identification of primary tumor phenotype from few CTCs isolated from peripheral blood, with the ultimate goal of introducing microfluidic and proteomic technologies to clinical cancer medicine. We plan to use an approach that integrates cutting edge technology and biology to address the isolation, purification, and proteomic sample preparation from CTCs isolated from cancer patients. The specific aim for this collaborative project is to design, fabricate and characterize a microfluidic chip for the preparation of samples for proteomic analysis starting from the captured CTCs, and to identify characteristic CTC phenotypes based on proteomic signatures.