Analytic and clinical validation of a prostate cancer-enhanced messenger RNA detection assay in whole blood as a prognostic biomarker for survival Journal Article


Authors: Danila, D. C.; Anand, A.; Schultz, N.; Heller, G.; Wan, M.; Sung, C. C.; Dai, C.; Khanin, R.; Fleisher, M.; Lilja, H.; Scher, H. I.
Article Title: Analytic and clinical validation of a prostate cancer-enhanced messenger RNA detection assay in whole blood as a prognostic biomarker for survival
Abstract: Background Biomarkers based on detecting prostate cancer (PCa)-specific transcripts in blood are associated with inferior outcomes, but their validation in a clinical context is lacking. Objective To determine whether detecting enhanced transcripts for PCa in whole blood using an analytically valid assay has prognostic significance relative to circulating tumor cell (CTC) enumeration. Design, setting, and participants The detection of KLK3, KLK2, HOXB13, GRHL2, and FOXA1 in whole blood by reverse transcription polymerase chain reaction (RT-PCR) was studied in 97 men with metastatic castration-resistant PCa (mCRPC) as a prognostic factor for overall survival. Intervention The 2.5 ml of blood was collected in PAXgene tubes for total RNA extraction and 7.5 ml for CTC enumeration from patients with progressive mCRPC. Outcome measurements and statistical analysis PCa-enriched genes were detected using a sensitive RT-PCR assay in whole blood from patients with mCRPC. Analytical validity of the assay was established in a clinical laboratory environment. The frequency of detecting transcripts was compared to CTC enumeration using CellSearch in an independent data set and survival associations were explored by concordance probability estimate (CPE). Results and limitations Two or more genes were detected by PCR in 53% of patients (51 of 97; 95% confidence interval [CI], 43-63%), and unfavorable CTC counts (five of more cells) were seen in 46% (45 of 97; 95% CI, 36-56%). Importantly, transcripts were detectable in 11 of 52 patients with favorable CTC counts (21%; 95% CI, 8-35%). Transcript detection predicted overall survival in a proportional hazards model. Significantly, the predictive accuracy of RT-PCR detection in combination with CTC enumeration had a CPE of 0.752 (standard error: 0.038), although this was limited by the number of patients evaluated. Conclusions This validated RT-PCR assay detecting prostate-specific RNA in whole blood is prognostic for survival and may assess patient risk in tandem with CellSearch CTC enumeration. Its clinical utility is being prospectively explored. © 2013 European Association of Urology.
Keywords: adult; cancer survival; clinical article; controlled study; aged; overall survival; cancer growth; outcome assessment; reverse transcription polymerase chain reaction; hepatocyte nuclear factor 3alpha; genetic transcription; validation study; tumor marker; prostate cancer; messenger rna; blood sampling; biomarker; circulating tumor cells; clinical laboratory; rna extraction; circulating tumor cell; castration resistant prostate cancer; human; male; priority journal; article; prostate-specific markers; cell counting; gene expression assay
Journal Title: European Urology
Volume: 65
Issue: 6
ISSN: 0302-2838
Publisher: Elsevier Science, Inc.  
Date Published: 2014-06-01
Start Page: 1191
End Page: 1197
Language: English
DOI: 10.1016/j.eururo.2013.07.006
PROVIDER: scopus
PUBMED: 23954088
PMCID: PMC4113474
DOI/URL:
Notes: Eur. Urol. -- Export Date: 2 June 2014 -- CODEN: EUURA -- Source: Scopus
Altmetric Score
MSK Authors
  1. Glenn Heller
    303 Heller
  2. Hans Gosta Lilja
    286 Lilja
  3. Raya Khanin
    38 Khanin
  4. Martin Fleisher
    244 Fleisher
  5. Howard Scher
    837 Scher
  6. Daniel C Danila
    84 Danila
  7. Aseem Anand
    45 Anand
  8. Nikolaus D Schultz
    206 Schultz
  9. Clifford Sung
    7 Sung
  10. Mingliang Wan
    4 Wan
  11. Charles S Dai
    1 Dai