Abstract
Abstract Background and Objectives: Over one million men undergo prostate biopsies annually in the US, a majority of them due to elevated serum PSA. More than half of the biopsies turn out to be negative for prostate cancer (CaP). The limitations of PSA test as well as the biopsy procedure have prompted to develop more precise CaP detection assays in urine- (e.g. PCA3, TMPRSS2-ERG) or blood (e.g. PHI, 4K). Here we describe the development and evaluation of a Urine CaP Marker Panel (UCMP) assay for sensitive and reproducible detection of CaP cells in post-digital rectal examination (post-DRE) urine. Methods: The cellular content of the post-DRE urine, was captured on translucent membrane for direct evaluation by microscopy and immunocytochemistry. The membranes placed on Cytoclear microscope slides were used for characterization of prostate epithelial cell by PSA and Prostein or tumor cells by ERG and AMACR protein markers. Immunostained cells were analyzed for quantitative and qualitative features and correlated with biopsy positive and negative status for malignancy. Results: Assay was optimized for single cell capture sensitivity and downstream evaluations by spiking known number of cells from established prostate cancer cell lines, LNCaP and VCaP, into cleared control urine. The cells captured from the post-DRE urine of subjects, obtained prior to biopsy procedure, were co-stained for ERG, AMACR, Prostein and PSA, rendering a whole cell based analysis and characterization. A feasibility cohort of 63 post-DRE urine specimens was assessed. Comparison of the UCMP results with the blinded biopsy results showed an assay sensitivity of 64% (16 of 25) and a specificity of 68.8% (22 of 32) for CaP detection by biopsy. Conclusion: This pilot study assessing a minimally invasive prostate cancer detection assay with single cell sensitivity cell-capture and characterization from the post-DRE urine holds promise for further development of this novel assay platform. Citation Format: Kristen Nickens, Amina Ali, Tatiana Scoggin, Shyh-han Tan, Lakshmi Ravindranath, David McLeod, David Tacha, Isabell Sesterhenn, Albert Dobi, Shiv Srivastava, Gyorgy Petrovics. Evaluation of a multiplex biomarker assay for the detection of prostate cancer cells in urine. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 623. doi:10.1158/1538-7445.AM2015-623
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