Clinical Research Papers:
Cell type specific gene expression analysis of prostate needle biopsies resolves tumor tissue heterogeneity
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Malte Krönig1, Max Walter1, Vanessa Drendel2, Martin Werner2, Cordula A. Jilg1, Andreas S. Richter3,4, Rolf Backofen3, David McGarry1, Marie Follo5, Wolfgang Schultze-Seemann1, Roland Schüle1
1Medical Center, University of Freiburg, Department of Urology, 79106 Freiburg, Germany
2Medical Center, University of Freiburg, Department of Pathology, 79106 Freiburg, Germany
3University of Freiburg, Department of Computer Science, 79110 Freiburg, Germany
4Max Planck Institute of Immunbiology and Epigenetics, 79108 Freiburg, Germany
5Medical Center, University of Freiburg, Department of Medicine I, 79106 Freiburg, Germany
Roland Schüle, e-mail: firstname.lastname@example.org
Keywords: prostate cancer, RNA detection, living cells, needle biopsy, gene expression, tumor heterogeneity
Received: August 06, 2014 Accepted: November 11, 2014 Published: December 01, 2014
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.
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