Intraoperational Prostate Loge Biopsies (iPROLOGX) After Radical Prostatovesiculectomy

Prostate Cancer Clinical Trial

— iPROLOGX  

Official title:

Intraoperational Prostate Loge Biopsies (iPROLOGX) After Radical Prostatovesiculectomy (RPVE) in Prostate Cancer (PCA) Patients for Molecular Tumor Marker Analysis

Clinical Trial Details — Status: Active, not recruiting

Administrative data

• NCT number: NCT02460861
• Other study ID #: MD-URO-00021
• Status: Active, not recruiting
• Phase: N/A
• First received: May 28, 2015
• Last updated: May 29, 2015
• Start date: November 2011
• Est. completion date: December 2015

Study information

• Verified date: May 2015
• Source: University of Magdeburg
• Contact: n/a
• Is FDA regulated: No
• Health authority: Germany: Ethics Commission
• Study type: Interventional

Clinical Trial Summary

This project is about the detection of occult tumor cells in surgical margins of radical prostatovesiculectomy by analysing the methylation status of Glutathione S-transferase P 1 (GSTP1). After gland excision specimens are obtained from 9 defined areas of the prostatic fossa. The biopsies are divided into two parts. One part used for histopathological analysis and the other part for moleculargenetic analysis. Results will be correlated e.g. with tumor stage, Gleason Score and prostate specific antigen (PSA).

The prostate-cancer-negative control group with bladder cancer.

Description:

This project is about the detection of occult tumor cells in surgical margins of radical prostatovesiculectomy by analysing the methylation status of Glutathione S-transferase P 1 (GSTP1). After gland excision specimens are obtained from 9 defined areas of the prostatic fossa. The biopsies are divided into two parts. One part used for histopathological analysis and the other part for moleculargenetic analysis. Results will be correlated e.g. with tumor stage, Gleason Score and prostate specific antigen (PSA).

The prostate-cancer-negative control group with bladder cancer.

DNA ISOLATION

DNA from biopsies stored by -80°C was isolated by using innuPREP DNA mini Kit (Analytik Jena, Jena, Germany) following protocol 1 of the manufacturer's instructions. DNA was eluted with 50 µl elution buffer. Concentration and purity were analysed by using Nanodrop 2000.

DNA BISULFITE MODIFICATION

DNA was modified by using EpiTect Bisulfite Kit (QIAGEN, Hilden, Germany) according to manufacturer's instructions. Samples were eluted once with 20 µl elution buffer.

QUANTITATIVE METHYLATION SPECIFIC PCR

Methylation status of GSTP1 is analysed by quantitative methylation-specific PCR (Q-MSP) using StepOnePlus Real-Time PCR System and StepOne Software v2.1 from Applied Biosystems (Darmstadt, Germany). Q-MSP was performed in duplicate analysing genes Actin and GSTP1. The primers' and testing probes' sequences used to amplify and detect hypermethylated GSTP1 were: 5'-AgTTgCgCggCgATTTC (forward primer), 5'-gCCCCAATACTAAATCACgACg (reverse primer) and 5'-CggTCgACgTTCggggTgTAgCg (taqman probe), labelled with fluorescence dye FAM. The primers' and testing probes' sequences used to amplify and detect Actin were: 5'-TggTgATggAggAggTTTAgTAAgT (forward primer), 5'-AACCAATAAAACCTACTCCTCCCTTAA (reverse primer),5'-ACCACCACCCAACACACAATAACAAACACA (taqman probe), labelled with fluorescence dye VIC.

The Q-MSP was carried out at 50°C for 2 min., 95°C for 15 min. followed by 50 cycles of 95°C for 1s and 60°C for 1 min. As a positive control bisulfite-converted DNA of DU145 and LNCap were used. Blank reactions with destillated water, which replaced DNA, served as negative control (NTC).

Recruitment information / eligibility

• Status: Active, not recruiting
• Enrollment: 279
• Est. completion date: December 2015
• Est. primary completion date: October 2013
• Accepts healthy volunteers: No
• Gender: Male
• Age group: N/A and older

Eligibility

Criteria for patients with prostate adenocarcinoma:

Inclusion Criteria

• sex: male

• diagnosis: prostate adenocarcinoma

• treatment: radical prostatovesiculectomy

• period of treatment: 11/30/2011 - 10/15/2013

Exclusion Criteria

• sex: female

• diagnosis: no prostate adenocarcinoma

• treatment: no radical prostatovesiculectomy

• period of treament: before 11/30/2011 or after 10/15/2013

Criteria for prostate adenocarcinoma negative control group:

Inclusion Criteria

• sex: male

• diagnosis: urothelial carcinoma

• treatment: cysto-prostatectomy

• period of treament: 12/14/2011 - 02/18/2014

Exclusion Criteria

• sex: female

• diagnosis: incidental prostate adenocarcinoma

• treatment: no cysto-prostatectomy

• period of treatment: before 12/14/2011 or after 02/18/2014

Study Design

Allocation: Non-Randomized, Intervention Model: Parallel Assignment, Masking: Open Label, Primary Purpose: Basic Science

Related Conditions & MeSH terms

• Prostate Cancer
• Prostatic Neoplasms

Intervention

Procedure:
• biopsies of the prostatic fossa in Magdeburg
intraoperative Open surgical biopsies of the prostatic fossa after prostatevesiculectomy in Magdeburg
• RPVE
Open Radical prostatovesiculectomy in Magdeburg
• ETRARP
Robotassisted Radical prostatovesiculectomy in Gronau
• CE
Open cystectomy in Magdeburg/Gronau
• biopsies of the prostatic fossa in gronau
intraoperative endoscopic robotassisted biopsies of the prostatic fossa after prostatevesiculectomy in Gronau

Locations

Country	Name	City	State
Germany	St. Antonius Hospital	Gronau	Nordrhein-Westfalen
Germany	Department of Urology, University Clinic Otto von Guericke University Magdeburg	Magdeburg	Sachsen-Anhalt

Sponsors (2)

Lead Sponsor	Collaborator
University of Magdeburg	St. Antonius Hospital Gronau

Country where clinical trial is conducted

Germany, 

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Methylation status of GSTP1	Methylation status of GSTP1 is analysed by quantitative methylation-specific PCR (Q-MSP)	2 years	No
Primary	Histopathology of prostate fossa biopsies	Histopathology of prostate fossa biopsies (Prostata cancer positive or negative)	2 years	No