Classifying indolent prostate cancer represents a significant clinical challenge. We investigated whether integrating data from different omic platforms could identify a biomarker panel with improved performance compared to individual platforms alone. DNA methylation, transcripts, protein and glycosylation biomarkers were assessed in a single cohort of patients treated by radical prostatectomy. Novel multiblock statistical data integration approaches were used to deal with missing data and modelled via stepwise multinomial logistic regression, or LASSO. After applying leave‐one‐out cross‐validation to each model, the probabilistic predictions of disease type for each individual panel were aggregated to improve prediction accuracy using all available information for a given patient. Through assessment of three performance parameters of area under the curve (AUC) values, calibration and decision curve analysis, the study identified an integrated biomarker panel which predicts disease type with a high level of accuracy, with Multi AUC value of 0.91 (0.89, 0.94) and Ordinal C‐Index (ORC) value of 0.94 (0.91, 0.96), which was significantly improved compared to the values for the clinical panel alone of 0.67 (0.62, 0.72) Multi AUC and 0.72 (0.67, 0.78) ORC. Biomarker integration across different omic platforms significantly improves prediction accuracy. We provide a novel multiplatform approach for the analysis, determination and performance assessment of novel panels which can be applied to other diseases. With further refinement and validation, this panel could form a tool to help inform appropriate treatment strategies impacting on patient outcome in early stage prostate cancer.

Background Between 20% and 35% of prostate cancer (PCa) patients who undergo treatment with curative intent (ie, surgery or radiation therapy) for localized disease will experience biochemical recurrence (BCR). Alterations in the insulin‐like growth factor (IGF) axis and PTEN expression have been implicated in the development and progression of several human tumors including PCa. We examined the expression of the insulin receptor (INSR), IGF‐1 receptor (IGF‐1R), PTEN, and AKT in radical prostatectomy tissue of patients who developed BCR post‐surgery. Methods Tissue microarrays (TMA) of 130 patients post‐radical prostatectomy (65 = BCR, 65 = non‐BCR) were stained by immunohistochemistry for INSR, IGF‐1R, PTEN, and AKT using optimized antibody protocols. INSR, IGF1‐R, PTEN, and AKT expression between benign and cancerous tissue, and different Gleason grades was assessed. Kaplan‐Meier survival curves were used to examine the relationship between proteins expression and BCR. Results INSR ( P < 0.001), IGF‐1R ( P < 0.001), and AKT ( P < 0.05) expression was significantly increased and PTEN ( P < 0.001) was significantly decreased in cancerous versus benign tissue. There was no significant difference in INSR, IGF‐1R, or AKT expression in the cancerous tissue of non‐BCR versus BCR patients ( P = 0.149, P = 0.990, P = 0.399, respectively). There was a significant decrease in PTEN expression in the malignant tissue of BCR versus non‐BCR patients ( P = 0.011). Combinational analysis of the tissue proteins identified a combination of decreased PTEN and increased AKT or increased INSR was associated with worst outcome. We found that in each case, our hypothesized worst group was most likely to experience BCR and this was significant for combinations of PTEN+INSR and PTEN+AKT but not PTEN+IGF‐1R ( P = 0.023, P = 0.028, P = 0.078, respectively). Conclusions Low PTEN is associated with BCR and this association is strongly modified by high INSR and high AKT expression. Measurement of these proteins could help inform appropriate patient selection for postoperative adjuvant therapy and prevent BCR.

You have accessJournal of UrologyProstate Cancer: Markers II1 Apr 2015MP6-20 THE ROLE OF THE INSULIN LIKE GROWTH FACTOR (IGF) AXIS IN PREDICTING BIOCHEMICAL RECURRENCE FOLLOWING RADICAL PROSTATECTOMY Kieran J. Breen, Noel M. Fitzgerald, Yue Fan, Susie Boyce, Amanda O'Neill, John M. Fitzpatrick, and R William Watson Kieran J. BreenKieran J. Breen More articles by this author , Noel M. FitzgeraldNoel M. Fitzgerald More articles by this author , Yue FanYue Fan More articles by this author , Susie BoyceSusie Boyce More articles by this author , Amanda O'NeillAmanda O'Neill More articles by this author , John M. FitzpatrickJohn M. Fitzpatrick More articles by this author , and R William WatsonR William Watson More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.267AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The insulin receptor (INSR) and IGF-1 receptor (IGF-1R) are key components of the Insulin-like Growth Factor (IGF) axis which has been implicated in prostate cancer development and progression. The tumour suppressor gene, PTEN, is a negative regulator of the PI3K pathway. Altered PTEN expression has been linked to lethal prostate cancer and is associated with the activation of PI3K and AKT. We examined the expression of the INSR, IGF-1R, PTEN and AKT in radical prostatectomy tissue of patients who developed biochemical recurrence (BCR) post-surgery. METHODS Tissue microarrays (TMA) of 130 patients post-radical prostatectomy (65 = BCR, 65 = non-BCR) were stained by immunohistochemistry for INSR, IGF-1R, PTEN and AKT using optimized antibody protocols. The TMAs were scored by two independent pathologists. INSR, IGF1-R, PTEN and AKT expression between benign and malignant tissue, and different Gleason grades was assessed using the Student t-test and ANOVA. Kaplan Meier survival curves were used to examine the relationship between proteins expression and BCR with p-values calculated using the log-rank test. RESULTS INSR (p<0.001), IGF-1R (p<0.001) and AKT (p<0.05) expression was significantly increased and PTEN (p<0.001) was significantly decreased in malignant versus benign tissue. There was no significant difference in INSR, IGF-1R or AKT expression in the malignant tissue of non-BCR versus BCR patients (p=0.149, p=0.991, p=0.399 respectively). There was a significant decrease in PTEN expression in the malignant tissue of BCR versus non-BCR patients (p=0.012). Combinational analysis of the tissue proteins, identified a combination of decreased PTEN and increased AKT or increased INSR was associated with worst outcome. We found that in each case our hypothesised worst group was most likely to experience BCR and this was significant for combinations of PTEN + INSR and PTEN + AKT but not PTEN + IGF-1R (p=0.023, p=0.028, p=0.078 respectively). CONCLUSIONS Low PTEN is associated with BCR and this association is strongly modified by high INSR and high AKT expression. Measurement of these proteins could help to inform appropriate patient selection for post-operative adjuvant therapy and prevent BCR. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e62-e63 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Kieran J. Breen More articles by this author Noel M. Fitzgerald More articles by this author Yue Fan More articles by this author Susie Boyce More articles by this author Amanda O'Neill More articles by this author John M. Fitzpatrick More articles by this author R William Watson More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Abstract Despite initial response to androgen ablation, patients with advanced disease relapse to develop castrate-resistant prostate cancer (CRPC) which is difficult to treat. We previously identified Serum Response Factor (SRF) as an important transcription factor in an in vitro model of CRPC. Since CRPC is associated with androgen receptor (AR) hypersensitivity and previous studies linked SRF with AR, we investigated the relationship between SRF and AR. DHT stimulation increased SRF transcriptional activity in both parental LNCaP cells and their castration-resistant Abl subline. To investigate AR involvement in this process, AR expression was down-regulated using siRNA, resulting in the abrogation of SRF induction post-DHT. Moreover, DHT stimulation failed to induce SRF transcriptional activity in the AR-negative PC346 DCC cells. However, when AR was over-expressed, SRF induction post-DHT was restored. To explore whether this was reciprocal, SRF was down-regulated by siRNA and AR transcriptional activity and protein expression were assessed. These experiments showed AR increased activity and expression following SRF knock-down, suggesting a negative feedback loop, which was confirmed by immunohistochemistry (IHC) in CRPC bone metastases (n=42), showing a negative association between SRF and AR positivity. This negative feedback was also supported by a computational model based on our experimental data. We next assessed cell proliferation following SRF inhibition and demonstrated that SRF inhibition by siRNA or CCG-1423 was as effective as AR inhibition in decreasing cell proliferation. Moreover concomitant inhibition of AR and SRF showed a synergistic effect. To confirm SRF relevance in clinical samples, we performed IHC staining for SRF on 94 primary tumours and 21 castration-resistant patients who received a transurethral resection of the prostate. This analysis showed that SRF nuclear staining was associated with castration-resistance with 95% of castration-resistant patients showing positive SRF staining versus only 50% of primary tumours. In addition SRF expression in bone metastases in a cohort of 38 patients who died of CRPC, was an independent predictor of survival from (a) prostate cancer diagnosis and (b) diagnosis with CRPC. Overall our data highlight the importance of SRF in CRPC and support SRF as a promising therapeutic target alone or in combination with current treatments. Citation Format: Maria Prencipe, Gillian O'Hurley, Amanda O'Neill, Dara Lundon, Lan K. Nguyen, Susie Boyce, Colm Morrissey, Helmut Klocker, Elaine W. Kay, William Gallagher, William Watson. Negative feedback loop between serum response factor and androgen receptor in castrate-resistant prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 4599. doi:10.1158/1538-7445.AM2014-4599

BACKGROUND:There are dilemmas associated with the diagnosis and prognosis of prostate cancer which has lead to over diagnosis and over treatment. Prediction tools have been developed to assist the treatment of the disease.METHODS:A retrospective review was performed of the Irish Prostate Cancer Research Consortium database and 603 patients were used in the study. Statistical models based on routinely used clinical variables were built using logistic regression, random forests and k nearest neighbours to predict prostate cancer stage. The predictive ability of the models was examined using discrimination metrics, calibration curves and clinical relevance, explored using decision curve analysis. The N = 603 patients were then applied to the 2007 Partin table to compare the predictions from the current gold standard in staging prediction to the models developed in this study.RESULTS:30% of the study cohort had non organ-confined disease. The model built using logistic regression illustrated the highest discrimination metrics (AUC = 0.622, Sens = 0.647, Spec = 0.601), best calibration and the most clinical relevance based on decision curve analysis. This model also achieved higher discrimination than the 2007 Partin table (ECE AUC = 0.572 & 0.509 for T1c and T2a respectively). However, even the best statistical model does not accurately predict prostate cancer stage.CONCLUSIONS:This study has illustrated the inability of the current clinical variables and the 2007 Partin table to accurately predict prostate cancer stage. New biomarker features are urgently required to address the problem clinician's face in identifying the most appropriate treatment for their patients. This paper also demonstrated a concise methodological approach to evaluate novel features or prediction models.

BACKGROUNDCastration-resistant prostate cancer (CRPC) represents a challenge to treat with no effective treatment options available. We recently identified serum response factor (SRF) as a key transcription factor in an in vitro model of castration resistance where we showed that SRF inhibition resulted in reduced cellular proliferation. We also demonstrated an association between SRF protein expression and CRPC in a cohort of castrate-resistant transurethral resections of the prostate (TURPS). The mechanisms regulating the growth of CRPC bone and visceral metastases have not been explored in depth due to the paucity of patient-related material available for analysis. In this study, we aim to evaluate SRF protein expression in prostate cancer (PCa) metastases, which has not previously been reported.METHODS AND RESULTSWe evaluated the nuclear tissue expression profile of SRF by immunohistochemistry in 151 metastatic sites from 42 patients who died of advanced PCa. No relationship between SRF nuclear expression and the site of metastasis was observed (P=0.824). However, a negative association between SRF nuclear expression in bone metastases and survival from (a) diagnosis with PCa (P=0.005) and (b) diagnosis with CRPC (P=0.029) was seen. These results demonstrate that SRF nuclear expression in bone metastases is associated with survival, with patients with the shortest survival showing high SRF nuclear expression and patients with the longest survival having low SRF nuclear expression.CONCLUSIONOur study indicates that SRF is a key factor determining patients' survival in metastatic CRPC and therefore may represent a promising target for future therapies. Prostate 74:306-313, 2014. (c) 2013 Wiley Periodicals, Inc.

BACKGROUND Accurate preoperative staging of prostate cancer (PCa) is important but current diagnostic methods cannot accurately determine extracapsular extension (ECE), resulting in the possible triage of patients towards a less appropriate arm of therapy. This has consequences to patient care and better methods of preoperatively determining ECE are required. METHODS We followed a biomarker development pathway and compared the preoperative serum expressions of VEGF-D, PEDF, IGF-I, IGFBP3, and CD14 in patients from the Irish Prostate Cancer Research Consortium (PCRC) with radical prostatectomy determined ECE against patients with nonECE. RESULTS The expression measurements of five proteins were fitted into a logistic regression model and backwards variable elimination methods were applied which resulted in a model with IGFBP3 and CD14 as the best combination biomarker panel. This panel was tested in an independent cohort of patients using an optimized multiplex electrochemiluminescence assay. Receiver operating characteristic curves were generated and the areas under the curve (AUC) were calculated as an estimation of prediction accuracy. The biomarker panel was validated with an AUC of 76.6%, and a sensitivity and specificity of 80% and 75% was obtained. CONCLUSIONS This is the first internally validated, preoperative serum biomarker panel that identifies ECE in patients with Gleason score 7 PCa with AUC 76.6%. The panel surpasses the routinely used diagnostic standards in accuracy and may help to improve preoperative cancer staging, better inform treatment options, and improve the referral patterns of patients with urgently treatable cancers towards more appropriate arms of therapy. Prostate 72:15231531, 2012. (c) 2012 Wiley Periodicals, Inc.

Eur J Clin Invest 2012AbstractBackground This study tested the hypothesis that surgical stress and the host response to this trauma trigger an inflammatory cascade in which the neutrophil plays a central role. We hypothesised that pre‐operative neutrophil migratory responses will correlate with post‐operative clinical outcome in our shock model of open‐heart surgery patients. We also tested the hypothesis that surface expression of adhesion molecules involved in the migratory process – CD11b, CD47 and CD99 – could be used to predict outcome. We believe that combining neutrophil migratory response, CD11b, CD47 and CD99 with the logistic European System for Cardiac Operative Risk Evaluation (EuroSCORE) will strengthen the power of the EuroSCORE not only in predicting post‐operative mortality but also other clinical endpoints.Materials and methods Neutrophils were isolated pre‐operatively from n = 31 patients undergoing open‐heart surgery and allowed to migrate across endothelial monolayers in response to N‐formyl‐methionine‐leucine‐phenylalanine (fMLP). Isolated neutrophils were also assessed for surface expression of CD11b, CD47 and CD99 in response to fMLP by flow cytometry. Post‐operative clinical parameters collected included days 1–5 white cell count and creatinine levels as well as intensive care unit (ICU) and post‐operative hospital stay.Results Pre‐operative surface expression of CD99 and CD47 correlates with post‐operative creatinine levels (P < 0·05), a measurement of renal injury. We also show that while the logistic EuroSCORE alone can be used as a predictor of ICU stay, when combined with pre‐operative CD99 surface expression, it improves its AUC value (0·794).Conclusion Immunological markers, specifically the ability of the neutrophil to migrate, combined with the logistic EuroSCORE lead to improved sensitivity and specificity to predict patient outcome.