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High-throughput screening of rare metabolically active tumor cells in pleural effusion and peripheral blood of lung cancer patients
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High-throughput screening of rare metabolically active tumor cells in pleural effusion and peripheral blood of lung cancer patients
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High-throughput screening of rare metabolically active tumor cells in pleural effusion and peripheral blood of lung cancer patients
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Journal Article
High-throughput screening of rare metabolically active tumor cells in pleural effusion and peripheral blood of lung cancer patients
2017
Overview
Malignant pleural effusion (MPE), the presence of malignant cells in pleural fluid, is often the first sign of many cancers and occurs in patients with metastatic malignancies. Accurate detection of tumor cells in pleural fluid is crucial because the presence of MPE denotes an advanced stage of disease and directs a switch in clinical managements. Cytology, as a traditional diagnostic tool, has limited sensitivity especially when tumor cells are not abundant, and may be confounded by reactive mesothelial cells in the pleural fluid. We describe a highly sensitive approach for rapid detection of metabolically active tumor cells in MPE via exploiting the altered glucose metabolism of tumor cells relative to benign cells. Metabolically active tumor cells with high glucose uptake, as evaluated by a fluorescent glucose analog (2-NBDG), are identified by high-throughput fluorescence screening within a chip containing 200,000 addressable microwells and collected for malignancy confirmation via single-cell sequencing. We demonstrate the utility of this approach through analyzing MPE from a cohort of lung cancer patients. Most candidate tumor cells identified are confirmed to harbor the same driver oncogenes as their primary lesions. In some patients, emergence of secondary mutations that mediate acquired resistance to ongoing targeted therapies is also detected before resistance is manifested in the clinical imaging. The detection scheme can be extended to analyze peripheral blood samples. Our approach may serve as a valuable complement to cytology in MPE diagnosis, helping identify the driver oncogenes and resistance-leading mutations for targeted therapies.
Publisher
National Academy of Sciences
Subject
4-Chloro-7-nitrobenzofurazan - analogs & derivatives
/ 4-Chloro-7-nitrobenzofurazan - metabolism
/ Antigens, Neoplasm - analysis
/ Biomarkers, Tumor - analysis
/ Carcinoembryonic Antigen - analysis
/ Carcinoembryonic Antigen - blood
/ Cytology
/ Deoxyglucose - analogs & derivatives
/ Genetics
/ High-Throughput Screening Assays - methods
/ Humans
/ Mutation
/ Pleural Effusion - diagnosis
/ Pleural Effusion - metabolism
/ Pleural Effusion, Malignant - blood
/ Pleural Effusion, Malignant - diagnosis
/ Pleural Effusion, Malignant - metabolism
/ Positron-Emission Tomography - methods
/ Tumors
