Isolation, Expansion and Characterization of Circulating Tumor Cells using Microfluidic Technologies.

Circulating tumor cells;Microfluidics;Lung cancer;Ex vivo expansion;Biomedical Engineering;Chemical Engineering;Oncology and Hematology;Engineering;Health Sciences;Chemical Engineering

Zhang, ZhuoGulari, Erdogan ;

University of Michigan

关键词: Circulating tumor cells;    Microfluidics;    Lung cancer;    Ex vivo expansion;    Biomedical Engineering;    Chemical Engineering;    Oncology and Hematology;    Engineering;    Health Sciences;    Chemical Engineering;   

Others  :  https://deepblue.lib.umich.edu/bitstream/handle/2027.42/120755/zzhuo_1.pdf?sequence=1&isAllowed=y

瑞士|英语

来源: The Illinois Digital Environment for Access to Learning and Scholarship

【 摘 要 】

Circulating tumor cells (CTCs) are the tumor cells shed from primary tumor, which enter the bloodstream and travel to distant anatomic sites to form metastasis. Detecting CTCs, as a means of ;;liquid biopsy”, allows monitoring cancer progression in real time and predicting therapeutic response. The major challenge that limits the clinical utility of CTCs, especially in early stage cancer patients, is their rarity; specifically, there are only 1-10 CTCs in one milliliter of whole blood. One way to overcome this critical limitation is to ex vivo expand CTCs through culturing. In this thesis, a microfluidic CTC capture device is optimized and tested for the capture and analysis of CTCs. Using a 3D, on chip co-culture model, CTCs were successfully expanded in 70% of the 50 early stage lung and esophageal cancer patients. Cultured CTCs were characterized with immunostaining, RNA profiling, mutation analysis and invasion assays. We found concordant TP53 mutation in CTCs and matched primary tumors. Next-generation sequencing further revealed mutations in additional genes. It was found that, patients whose CTCs exhibited the greatest capacity to expand had earlier recurrence. In one of the patient samples, expanded CTCs implanted in mice resulted in a xenograft tumor, demonstrating the expanded CTCs’ ability to metastasize. Genes related to EMT and tumor microenvironment were enriched in the xenograft. In addition, building upon this co-culture model, CTCs from one of the ALK positive metastatic lung cancer patients were isolated and cultured. The cultured CTCs harbored the concordant EML4-ALK rearrangement as the tumor biopsy specimen and further served as an in vitro model for drug testing. Taken together, this study demonstrated that CTCs from early stage lung cancer are tumorigenic and mirror the phenotypic and genotypic status of primary tumors. Ex vivo culturing of CTCs will make a significant impact in the era of personalized medicine. It will bring about opportunities for individualized drug screening, such as predicting treatment response to targeted therapies and the emergence of acquired drug resistance. Cultured CTCs will also serve as tools for understanding metastatic spread of cancer cells.