SORTING AND PROTEOMIC CHARACTERIZATION OF CIRCULATING TUMOR CELLS

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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.