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Functional characterization of circulating tumor cells with a prostate-cancer-specific microfluidic device.
- Source :
-
PloS one [PLoS One] 2012; Vol. 7 (4), pp. e35976. Date of Electronic Publication: 2012 Apr 27. - Publication Year :
- 2012
-
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.
- Subjects :
- Base Sequence
Biological Assay
Cell Line, Tumor
Computer Simulation
Equipment Design
Humans
Male
Molecular Imaging
Molecular Sequence Data
Neoplasm Metastasis
Neoplastic Cells, Circulating drug effects
Neoplastic Cells, Circulating metabolism
Oncogene Proteins, Fusion metabolism
Organ Specificity drug effects
Point Mutation genetics
Prostatic Neoplasms metabolism
Protein Binding drug effects
Receptors, Androgen genetics
Taxoids pharmacology
Tubulin metabolism
Microfluidic Analytical Techniques methods
Neoplastic Cells, Circulating pathology
Prostatic Neoplasms pathology
Subjects
Details
- Language :
- English
- ISSN :
- 1932-6203
- Volume :
- 7
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- PloS one
- Publication Type :
- Academic Journal
- Accession number :
- 22558290
- Full Text :
- https://doi.org/10.1371/journal.pone.0035976