title: High-throughput screening of rare metabolically active tumor cells in pleural effusion and peripheral blood of lung cancer patients.
abstract: 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.
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
authors.forename: Zhuo
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Shanghai Lung Cancer Center, Shanghai Chest Hospital, Shanghai, 200030, China.
authors.forename: Ziming
authors.initials: Z
authors.lastname: Li
authors.affiliation: NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, CA 91125.|Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125.
authors.forename: Jungwoo
authors.initials: J
authors.lastname: Kim
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China.
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Shanghai Municipal Hospital of Traditional Chinese Medicine, Shanghai, 200071, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, CA 91125.|Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125.|Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: James R
authors.initials: JR
authors.lastname: Heath
authors.affiliation: NanoSystems Biology Cancer Center, California Institute of Technology, Pasadena, CA 91125; weiwei@mednet.ucla.edu lushun@csco.org.cn qihuishi@sjtu.edu.cn.|Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095.
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.affiliation: Shanghai Lung Cancer Center, Shanghai Chest Hospital, Shanghai, 200030, China; weiwei@mednet.ucla.edu lushun@csco.org.cn qihuishi@sjtu.edu.cn.
authors.forename: Shun
authors.initials: S
authors.lastname: Lu
authors.affiliation: Key Laboratory of Systems Biomedicine (Ministry of Education), School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200240, China; weiwei@mednet.ucla.edu lushun@csco.org.cn qihuishi@sjtu.edu.cn.
authors.forename: Qihui
authors.initials: Q
authors.lastname: Shi
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000378:metabolism; D000072283:A549 Cells; D000951:Antigens, Neoplasm / Q000032:analysis / Q000097:blood; D014408:Biomarkers, Tumor / Q000032:analysis / Q000097:blood; D002272:Carcinoembryonic Antigen / Q000032:analysis / Q000097:blood; D045744:Cell Line, Tumor; D003847:Deoxyglucose / Q000031:analogs & derivatives / Q000378:metabolism; D003937:Diagnosis, Differential; D005947:Glucose / Q000378:metabolism; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007962:Leukocytes; D008175:Lung Neoplasms / Q000097:blood / Q000175:diagnosis* / Q000378:metabolism*; D010996:Pleural Effusion / Q000097:blood / Q000175:diagnosis* / Q000378:metabolism*; D016066:Pleural Effusion, Malignant / Q000097:blood / Q000175:diagnosis* / Q000378:metabolism*; D049268:Positron-Emission Tomography / Q000379:methods
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D002272:Carcinoembryonic Antigen; D003847:Deoxyglucose; D009327:4-Chloro-7-nitrobenzofurazan; D005947:Glucose; C098340:2-(N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino)-2-deoxyglucose
keywords: CTC; diagnosis; glucose uptake; lung cancer; pleural effusion