Abstract
A lack of cell surface markers for the specific identification, isolation and subsequent analysis of living prostate tumor cells hampers progress in the field. Specific characterization of tumor cells and their microenvironment in a multi-parameter molecular assay could significantly improve prognostic accuracy for the heterogeneous prostate tumor tissue. Novel functionalized gold-nano particles allow fluorescence-based detection of absolute mRNA expression levels in living cells by fluorescent activated flow cytometry (FACS). We use of this technique to separate prostate tumor and benign cells in human prostate needle biopsies based on the expression levels of the tumor marker alpha-methylacyl-CoA racemase (AMACR). We combined RNA and protein detection of living cells by FACS to gate for epithelial cell adhesion molecule (EPCAM) positive tumor and benign cells, EPCAM/CD45 double negative mesenchymal cells and CD45 positive infiltrating lymphocytes. EPCAM positive epithelial cells were further sub-gated into AMACR high and low expressing cells. Two hundred cells from each population and several biopsies from the same patient were analyzed using a multiplexed gene expression profile to generate a cell type resolved profile of the specimen. This technique provides the basis for the clinical evaluation of cell type resolved gene expression profiles as pre-therapeutic prognostic markers for prostate cancer.
Highlights
In men, prostate cancer is the most commonly diagnosed cancer and is the second leading cause of cancer related deaths [1]
We can conclude that alpha-methylacyl-CoA racemase (AMACR) mRNA expression level-based separation of epithelial cells generates cell populations that closely resemble tumor and benign prostate cells, with respect to their gene expression profile
The majority of the above data is acquired from bulk tissue analysis and may not represent the expression levels found in a cell type resolved analysis
Summary
Prostate cancer is the most commonly diagnosed cancer and is the second leading cause of cancer related deaths [1]. For 2012, the incidence was estimated to be 238,000 and the mortality to be 29,000 in the USA. Even though 5-year survival rates exceed 98%, appropriate risk adapted therapies remain a challenge in identifying and separating patients with low and highrisk cancer [1]. Data from one of the largest PSA screening trials (European Randomized Study of Screening for Prostate Cancer: ERSPC) [2] of more than 180,000 men estimated that rates for over-treatment are up to 50% in low risk groups. High mortality rates in the high-risk group clearly demonstrate a need for improved therapeutic strategies
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