Single Cell Characterization of Prostate Cancer-Circulating Tumor Cells
Abstract
Circulating tumor cells (CTC) mediate metastatic spread of many solid tumors. Our proposal exploited a new technology, MagSweeper, for isolation of CTCs from patients with prostate cancer. MagSweeper is an automated immunomagnetic separation technology that gently extracts CTCs from whole blood, allowing the possibility of characterization of the cells molecularly. We have now successfully demonstrated that the MagSweeper isolates completely purified CTCs while preserving the integrity and viability of these fragile cells and eliminating contamination from nonspecific adsorption or entrapment of blood cells. We have optimized the isolation of spiked LNCaP cells from the blood of healthy donors and then from the isolated cells performed whole transcriptome mRNA-Seq. MagSweeper can effectively isolate these cells with a high capture efficiency and purity. In addition, the MagSweeper isolation process does not significantly perturb the transcriptional signature of isolated single cells. We also have been able to isolate CTCs from patients with metastatic prostate cancer, and the numbers of cells isolated were comparable to CellSearch, a commercial FDA approved CTC counting technology. Although the quality of RNA from the CTCs has showed signs of degradation, we have been able to generate whole transcriptome profiles from single cells from patients using mRNA-Seq. Expressed transcripts showed the cells were of prostatic origin and reflected known aspects of prostate cancer biology. These results demonstrate that the MagSweeper provides access to intact CTCs and that it is possible to extract biologically and clinically relevant information from molecular analysis of single CTCs.
Document Details
- Document Type
- Technical Report
- Publication Date
- Sep 01, 2013
- Accession Number
- ADA596639
Entities
People
- James D Brooks
Organizations
- Stanford University