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Single Cell Characterization of Prostate Cancer Circulating Tumor Cells

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Annual rept. 1 Jul 2010-30 Jun 2011

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Our proposal exploits a new technology, MagSweeper, for isolation of Circulating Tumor Cells CTCs from patients with prostate cancer. MagSweeper is an automated immunomagnetic separation technology that gently extracts CTCs from whole blood, allowing the possibility of molecular characterization of the isolated cells. In this grant, we proposed 3 specific aims and made significant progress in all of them. For the first aim we now have optimized a protocol that captures a high EpCAM expressing cell line LNCaP and a low EpCAM cell line PC3 with high purity and efficiency. For the second aim, we applied the optimized protocol on isolating circulating tumor cells from 40 prostate cancer patients and found CTCs in 15 of those patients. To demonstrate purity and specificity of the isolated CTCs, we used RT PCR to demonstrate the presence of Prostate Specific Antigen PSA and Androgen Receptor AR transcripts in a subset of these cells. Finally, we have begun applying nextgen sequencing technologies to develop a whole transcriptome sequencing protocol for pooled and single CTCs. In the second year of funding, we plan to increase our sample size of patients with metastatic disease and attempt to isolate CTCs from patients with localized prostate cancer.

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  • Medicine and Medical Research

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