title: Prostate cancer marker panel with single cell sensitivity in urine.
abstract: BACKGROUND
  Over one million men undergo prostate biopsies annually in the United States, a majority of whom due to elevated serum PSA. More than half of the biopsies turn out to be negative for prostate cancer (CaP). The limitations of both the PSA test and the biopsy procedure have led to the development for 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 the 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 a translucent filter membrane, which is placed on Cytoclear slides for direct evaluation by microscopy and immuno-cytochemistry (ICC). Cells captured on the membrane were assayed for PSA and Prostein expression to identify prostate epithelial cells, and for ERG and AMACR to identify prostate tumor cells. Immunostained cells were analyzed for quantitative and qualitative features and correlated with biopsy positive and negative status for malignancy.
  
  
  RESULTS
  The assay was optimized for single cell capture sensitivity and downstream evaluations by spiking a known number of cells from established CaP cell lines, LNCaP and VCaP, into pre-cleared control urine. The cells captured from the post-DRE urine of subjects, obtained prior to biopsy procedure, were co-stained for ERG, AMACR (CaP specific), and Prostein or PSA (prostate epithelium specific) 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 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.
  
  
  CONCLUSIONS
  This pilot study assessing a minimally invasive CaP 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. Prostate 75: 969-975, 2015. © 2015 The Authors. The Prostate, published by Wiley Periodicals, Inc.
authors: Department of Surgery, Center for Prostate Disease Research, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
authors: Kristen P
authors: KP
authors: Nickens
authors: Amina
authors: A
authors: Ali
authors: Tatiana
authors: T
authors: Scoggin
authors: Shyh-Han
authors: SH
authors: Tan
authors: Lakshmi
authors: L
authors: Ravindranath
authors: David G
authors: DG
authors: McLeod
authors: Albert
authors: A
authors: Dobi
authors: David
authors: D
authors: Tacha
authors: Isabell A
authors: IA
authors: Sesterhenn
authors: Shiv
authors: S
authors: Srivastava
authors: Gyorgy
authors: G
authors: Petrovics
mesh_terms: D014408:Biomarkers, Tumor / Q000652:urine*; D045744:Cell Line, Tumor; D015331:Cohort Studies; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007610:Kallikreins / Q000652:urine; D008297:Male; D008565:Membrane Proteins / Q000652:urine; D010865:Pilot Projects; D017430:Prostate-Specific Antigen / Q000652:urine; D011471:Prostatic Neoplasms / Q000652:urine*; D019998:Racemases and Epimerases / Q000652:urine; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D015534:Trans-Activators / Q000652:urine; D000071230:Transcriptional Regulator ERG
chemical_list: D014408:Biomarkers, Tumor; C072312:ERG protein, human; D008565:Membrane Proteins; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG; C422655:prostein; C549870:KLK3 protein, human; D007610:Kallikreins; D017430:Prostate-Specific Antigen; D019998:Racemases and Epimerases; C108352:alpha-methylacyl-CoA racemase
keywords: biomarker panel; cell capture; prostate cancer; urine