Eur Urol Suppl 2011;10(2):176 scores generated by IHC-MARK correlated excellent to manual assessment. Patients with tumors expressing high levels of MSMB had a signifcantly reduced risk for biochemical recurrence after radical prostatectomy (HR= 0.468; 95% CI 0.394-0.556; p<0.001). Multivariate analysis adjusted for clinicopathological parameters revealed that MSMB expression was an independent predictor of decreased risk of recurrence (HR= 0.710; 95% CI 0.578-0.872; p<0.001). We found no correlation between CRISP3 expression and biochemical recurrence. Conclusions: In the current study, we applied a novel image analysis on a large independent cohort, and successfully verifed that MSMB is a strong independent factor predicting favorable outcome after radical prostatectomy for localized prostate cancer. The image analysis assessment algorithm, IHC-MARK, was sensitive and selective in tumor cells identifcation and scoring, and we believe that automated image analysis is a clinically useful tool for biomarker discovery and evaluation. 527 integrated diagnostiC Methods for deteCtion of MultiPle gene rearrangeMents in Prostate CanCer tissue sPeCiMens Cortez C. 1 , Svensson M. 2 , Nagle R. 1 , Garsha K. 1 , Nagy D. 1 , Riley J. 1 , Sathyanarayana U. 1 , Smith K. 1 , Pozarowski K. 1 , Christopherson K. 1 , Yun S. 1 , Otter M. 1 , Dittamore R. 1 , Palanisamy N. 3 , Demichelis F. 2 , Rubin M. 2 , Grogan T. 1 , Miller P. 1 , Pestano G. 1 1 Ventana Medical Systems, Tucson, United States of America, 2 Weill Cornell Medical College, Pathology and Laboratory Medicine, New York, United States of America, 3 University of Michigan, Michigan Center for Translational Pathology, Ann Arbor, United States of America introduction & objectives: Identifcation of patients who will beneft from therapy is one of the more diffcult questions in prostate cancer disease management. Approximately 60% of men with diagnosed prostate cancer will have an ETS rearranged tumor foci. The prostate cancer gene fusions identifed thus far are characterized by the fusion of 5’ genomic regulatory elements (commonly androgen regulated), such as TMPRSS2, with the ETS family of transcription factors which include ERG, ETV1, ETV4, and ETV5. About 40-50% of prostate cancers have the TMPRSS2/ERG gene fusion which leads to the over expression of oncogenic transcription factors. Materials & Methods: The goal of this study was to develop a novel, integrated diagnostic testing method that accounts for tissue heterogeneity including multiple gene rearrangements in single transformed nuclei. We selected a method that sequentially refexes from an initial H&E to ERG immunohistochemistry (IHC), and fnally to a quantum dot (Life Technologies) based FISH probes for the detection of multiple gene rearrangements in prostate cancer. To this end, probes specifc to the ETS gene rearrangements including, 3’ and 5’ ERG, TMPRSS2, NDRG1, ETV1, and ETV4, were detected with up to four different quantum dot bioconjugates simultaneously in single nuclei. results: We have shown sensitive and specifc detection of gene rearrangements with this testing method in the xenograft models, VCaP, H660, and LNCaP, as well as in samples from prostate needle biopsies and radical prostatectomies. In 6 of 88 cases, the ERG IHC was diagnostic of PIN (prostatic intraepithelial neoplasia) that was missed on initial examination of the H&E stain. Further, the four color FISH assay inclusive of ERG gene rearrangements was confrmatory of ERG IHC reactivity. In addition, we also clearly demonstrated instances of multiple gene rearrangements in the 5’ and 3’ ends on TMPRSS2 and NDRG1 in the same cancer nuclei, but not in benign nuclei from the same tissue. The signal patterns associated with various genomic events assessed were: no rearrangement (normal), translocation through insertion, and translocation through deletion. Conclusions: In summary, we propose that the ERG antibody is likely to be a key component in a diagnostic PIN IHC cocktail, that follows the initial H&E, and may be refexed to a multiplexed quantum dot FISH assay that incorporates the detection of prevalent gene rearrangements. This method may be useful as an aid in detecting clinically relevant diagnostic markers, since multiple gene rearrangements in the same cell may be identifed early in prostate cancer progression. Studies are in progress to evaluate this testing strategy for prognostic value in assessing prostate cancer progression in selected prostate cancer and biopsy cohorts. At time of submission, assay is not approved for use in the US. 528 re-ClassifiCation strategies in Men with low risk Prostate CanCer: trus guided bioPsy versus teMPlate Prostate MaPPing Barzell W. 1 , Cathcart P. 2 , Moore C. 2 , Melamed M. 3 , Ahmed H. 2 , Emberton M. 2 1 FSU College of Medicine, Dept. of Urology, Sarasota, Florida, United States of America, 2 UCLH/UCL Comprehensive Biomedical Research Centre, UCL Division of Surgery and Interventional Science, London, United Kingdom, 3 FSU College of Medicine, Dept. of Urology, Sarosota, Florida, United States of America introduction & objectives: Active surveillance (AS) is increasingly recommended to reduce over-treatment in patients with favourable-risk prostate cancer. Suitability for AS is often assessed by re-applying an additional biopsy strategy. We investigate the utility of repeat TRUS biopsy versus template prostate mapping (TPM) in a group of men being considered for AS. Materials & Methods: Between 2002 and 2009, 129 men with favourable-risk cancer underwent simultaneous repeat TRUS biopsy together with TPM. Gleason grade and burden were compared between the two biopsy strategies to assess eligibility for AS. Cancer signifcance was attributed using 4 commonly used defnitions. results: Only 30% of men identifed as having low-risk prostate cancer prior to study entry had cancer detected on repeat TRUS biopsy. In contrast, 80% had cancer detected on TPM (p<0.001). Irrespective of biopsy strategy, eligibility to active surveillance was highly dependant on the defnition of clinical signifcance used; When TPM was used as the re-classifcation method, 26% to 80% were eligible for AS in contrast to 79% to 95% that were deemed eligible after TRUS biopsy. Overall, TRUS biopsy ‘missed’ between 55% and 75% of clinically signifcant prostate cancers. Agreement between TRUS biopsy and TPM was at best only fair (kappa coeffcient 0.12-0.40). TRUS Biopsy TMP Cancer Detection 30% 80% Eligibility for AS Epstein criteria 79% 26% Conclusions: When evaluating suitability for active surveillance, 3-D template mapping would appear to represent a more accurate test to identify which men have low-risk prostate cancer. 529 a noMograM to PrediCt uPgrading or uPstaging in Patients that fit Conventional aCtive surveillanCe Criteria Sooriakumaran P. 1 , Srivastava A. 1 , Christos P. 2 , Grover S. 1 , Leung R. 1 , Tewari A.K. 1 1 Weill Cornell Medical College, Dept. of Urology, New York, United States of America, 2 Weill Cornell Medical College, Dept. of Statistics, New York, United States of America introduction & objectives: Low risk prostate cancer patients clinically eligible for active surveillance can also be managed surgically. We evaluated the pathologic outcomes for this cohort that was treated by radical prostatectomy by a single surgeon at our institution and devised a nomogram to predict which of this cohort are at risk of upgrading or upstaging that would have then made them ineligible for active surveillance. Materials & Methods: 750 patients treated by radical prostatectomy from Jan 2005-present fulflled conventional active surveillance criteria (PSA<10, Gleason sum 6, ≤cT2a) and formed the study cohort. Preoperative data on PSA, clinical stage, number of biopsy cores taken, number of positive cores, maximum percent cancer in any core, presence of high grade prostatic intraepithelial neoplasia (HGPIN), and prostate volume was available. The radical prostatectomy specimens were graded and staged, and any upgrading (Gleason sum >6) and/or upstaging (≥pT2b) was classed as ‘worsening prognosis’ as these factors would be considered exclusion criteria for active surveillance if known prior to treatment. Univariate analyses using t-tests, Wilcoxon rank-sum tests, and chi-square tests were performed to determine predictors of worsening prognosis from the list of preoperative variables. A multivariable logistic regression model was used to develop the worsening prognosis predictive nomogram. results: 297/750 (39.6%) patients were upgraded to ≥Gleason 7 at fnal pathology; 569/750 (75.9%) were upstaged to ≥pT2b; overall, 597/750 (79.6%) had either upgrading or upstaging and would have been ineligible for active surveillance if known prior to treatment. Univariate analysis found that only HGPIN had little predictive power for upgrading and/or upstaging, and thus all other variables were included in the multivariate model. The nomogram to predict worsening prognosis was reasonably discriminatory between patients who would have been ineligible for active surveillance and those that remained eligible (bootstrap corrected c-index of 0.65). Conclusions: Four out of fve patients deemed eligible for active surveillance based on conventional criteria have worse prognostic factors in terms of histology or tumor bulk when subjected to radical prostatectomy. We suggest the use of a nomogram we have devised to adequately counsel primary prostate cancer patients deemed clinically eligible for active surveillance. 530 radial distanCe of extraProstatiC extension Correlates with bioCheMiCal reCurrenCe after radiCal ProstateCtoMy Van Oort I.M. 1 , Van Veggel B.A.M.H. 1 , Hulsbergen-Van De Kaa C.A. 2 , Kiemeney L.A.L.M. 3 , Witjes J.A. 1 1 Radboud University Nijmegen Medical Centre, Dept. of Urology, Nijmegen, The Netherlands, 2 Radboud University Nijmegen Medical Centre, Dept. Pathology, Nijmegen, The Netherlands, 3 Radboud University Nijmegen Medical Centre, Dept. of Epidemiology, Biostatistics and HTA, Nijmegen, The Netherlands introduction & objectives: No consensus exists on how to substage extraprostatic extension (EPE) in prostate cancer. We compared radial distance to other methods, and correlated them to biochemical recurrence (BCR) after radical prostatectomy.