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Open Access Highly Accessed Technical advance

Isolation and genomic analysis of circulating tumor cells from castration resistant metastatic prostate cancer

Mark Jesus M Magbanua1, Eduardo V Sosa1, Janet H Scott1, Jeff Simko25, Colin Collins3, Dan Pinkel45, Charles J Ryan15 and John W Park156*

Author Affiliations

1 Department of Medicine, Division of Hematology/Oncology, University of California San Francisco (UCSF), San Francisco, CA, USA

2 Department of Pathology and Urology, UCSF, San Francisco, CA, USA

3 Vancouver Prostate Center and Department of Urologic Sciences, University of British Columbia, Vancouver, BC, Canada

4 Department of Laboratory Medicine, UCSF, San Francisco, CA, USA

5 Helen Diller Family Comprehensive Cancer Center, UCSF, San Francisco, CA, USA

6 Division of Hematology/Oncology, University of California San Francisco, 2340 Sutter St. Box 1387, San Francisco, CA 94115, USA

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BMC Cancer 2012, 12:78  doi:10.1186/1471-2407-12-78

Published: 28 February 2012

Abstract

Background

The number of circulating tumor cells (CTCs) in metastatic prostate cancer patients provides prognostic and predictive information. However, it is the molecular characterization of CTCs that offers insight into the biology of these tumor cells in the context of personalized treatment.

Methods

We developed a novel approach to isolate CTCs away from hematopoietic cells with high purity, enabling genomic analysis of these cells. The isolation protocol involves immunomagnetic enrichment followed by fluorescence activated cell sorting (IE/FACS). To evaluate the feasibility of isolation of CTCs by IE/FACS and downstream genomic profiling, we conducted a pilot study in patients with metastatic castration resistant prostate cancer (CRPC). Twenty (20) sequential CRPC patients were assayed using CellSearchâ„¢. Twelve (12) patients positive for CTCs were subjected to immunomagnetic enrichment and fluorescence activated cell sorting (IE/FACS) to isolate CTCs. Genomic DNA of CTCs was subjected to whole genome amplification (WGA) followed by gene copy number analysis via array comparative genomic hybridization (aCGH).

Results

CTCs from nine (9) patients successfully profiled were observed to have multiple copy number aberrations including those previously reported in primary prostate tumors such as gains in 8q and losses in 8p. High-level copy number gains at the androgen receptor (AR) locus were observed in 7 (78%) cases. Comparison of genomic profiles between CTCs and archival primary tumors from the same patients revealed common lineage. However, high-level copy number gains in the AR locus were observed in CTCs, but not in the matched archival primary tumors.

Conclusions

We developed a new approach to isolate prostate CTCs without significant leukocyte admixture, and to subject them to genome-wide copy number analysis. Our assay may be utilized to explore genomic events involved in cancer progression, e.g. development of castration resistance and to monitor therapeutic efficacy of targeted therapies in clinical trials in a relatively non-invasive manner.

Keywords:
Circulating tumor cells; Castration resistant prostate cancer; Copy number analysis; Array comparative genomic hybridization; Androgen receptor