Title |
Functional Characterization of Circulating Tumor Cells with a Prostate-Cancer-Specific Microfluidic Device
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Published in |
PLOS ONE, April 2012
|
DOI | 10.1371/journal.pone.0035976 |
Pubmed ID | |
Authors |
Brian J. Kirby, Mona Jodari, Matthew S. Loftus, Gunjan Gakhar, Erica D. Pratt, Chantal Chanel-Vos, Jason P. Gleghorn, Steven M. Santana, He Liu, James P. Smith, Vicente N. Navarro, Scott T. Tagawa, Neil H. Bander, David M. Nanus, Paraskevi Giannakakou |
Abstract |
Cancer metastasis accounts for the majority of cancer-related deaths owing to poor response to anticancer therapies. Molecular understanding of metastasis-associated drug resistance remains elusive due to the scarcity of available tumor tissue. Isolation of circulating tumor cells (CTCs) from the peripheral blood of patients has emerged as a valid alternative source of tumor tissue that can be subjected to molecular characterization. However, issues with low purity and sensitivity have impeded adoption to clinical practice. Here we report a novel method to capture and molecularly characterize CTCs isolated from castrate-resistant prostate cancer patients (CRPC) receiving taxane chemotherapy. We have developed a geometrically enhanced differential immunocapture (GEDI) microfluidic device that combines an anti-prostate specific membrane antigen (PSMA) antibody with a 3D geometry that captures CTCs while minimizing nonspecific leukocyte adhesion. Enumeration of GEDI-captured CTCs (defined as intact, nucleated PSMA+/CD45- cells) revealed a median of 54 cells per ml identified in CRPC patients versus 3 in healthy donors. Direct comparison with the commercially available CellSearch® revealed a 2-400 fold higher sensitivity achieved with the GEDI device. Confocal microscopy of patient-derived GEDI-captured CTCs identified the TMPRSS2:ERG fusion protein, while sequencing identified specific androgen receptor point mutation (T868A) in blood samples spiked with only 50 PC C4-2 cells. On-chip treatment of patient-derived CTCs with docetaxel and paclitaxel allowed monitoring of drug-target engagement by means of microtubule bundling. CTCs isolated from docetaxel-resistant CRPC patients did not show any evidence of drug activity. These measurements constitute the first functional assays of drug-target engagement in living circulating tumor cells and therefore have the potential to enable longitudinal monitoring of target response and inform the development of new anticancer agents. |
X Demographics
Geographical breakdown
Country | Count | As % |
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United States | 1 | 50% |
Unknown | 1 | 50% |
Demographic breakdown
Type | Count | As % |
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Scientists | 1 | 50% |
Members of the public | 1 | 50% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 4 | 2% |
Switzerland | 1 | <1% |
Malaysia | 1 | <1% |
France | 1 | <1% |
Italy | 1 | <1% |
Lithuania | 1 | <1% |
Netherlands | 1 | <1% |
Saudi Arabia | 1 | <1% |
United Kingdom | 1 | <1% |
Other | 2 | <1% |
Unknown | 210 | 94% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 59 | 26% |
Researcher | 33 | 15% |
Student > Master | 24 | 11% |
Student > Bachelor | 24 | 11% |
Other | 14 | 6% |
Other | 37 | 17% |
Unknown | 33 | 15% |
Readers by discipline | Count | As % |
---|---|---|
Engineering | 64 | 29% |
Agricultural and Biological Sciences | 42 | 19% |
Medicine and Dentistry | 31 | 14% |
Biochemistry, Genetics and Molecular Biology | 23 | 10% |
Chemistry | 11 | 5% |
Other | 15 | 7% |
Unknown | 38 | 17% |