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MP66-05 PRETREATMENT SYSTEMIC INFLAMATORY RESPONSE PARAMETERS DO NOT PREDICT THE OUTCOME IN MEN WITH PROSTATE CANCER UNDERGOING RADICAL PROSTATECTOMY
THE JOURNAL OF UROLOGYâ
Vol. 193, No. 4S, Supplement, Monday, May 18, 2015
therapeutic response. The goal of this study was to evaluate and genetically characterize exosome derived RNA (exoRNA) isolated from blood of metastatic CRPC patients. METHODS: Whole blood samples from 18 consented clinically annotated mCRPC patients and 1 normal control were collected. Exosomes were isolated with ultracentrifugation and exoRNA extracted. Following library prep, paired-end sequencing was performed using Illumina Hi-Seq 2000. A bioinformatics pipeline was used for data prepossessing including alignment, duplicate removal, normalization and variant calling. Visualization and differential analyses were performed with SNP & Variation Suite v8.x. RESULTS: In exoRNA there is evidence of extensive chromosomal rearrangement resulting in a myriad of gene fusions and isoforms. Through preliminary analyses we identified 39 genes commonly expressed in the exosomes of these mCRPC patients. These include PDPK1, USP9X, MAGI2, HMGA2 and PTGFR all of which have been previously expressed in prostate cancer tissue. A diverse variety of lcnRNA, ncRNA and miRNA were also identified in circulating exosomes. Validation of these alterations and additional analyses evaluating translocations and splice variants, PCR validation, and/or direct tumor based nucleic acid assays is ongoing. CONCLUSIONS: These preliminary analyses of circulating exoRNA have identified gene expression signatures of several prostate cancer associated transcripts. Ultimately, the identification of PCa associated transcripts in plasma derived exosomes provides evidence that exosomes and exosomal cargo may serve as biomarker in CRPC patients. Exosomes and exoRNA may provide otherwise unattainable insight into tumor evolution and disease progression. Additional studies evaluating the clinical relevance and prognostic value of exosomal RNA will be critical for biomarker development. Source of Funding: Tulane Cancer Center
MP66-04 CABAZITAXEL INHIBITS THE PROLIFERATION OF HUMAN CASTRATION-REFRACTORY PROSTATE CANCER CELLS IN VITRO AND ENHANCES THE ANTI-TUMOR PROPERTIES OF THE ANGIO-INHIBITORY PIGMENT EPITHELIUM-DERIVED FACTOR IN VIVO WITH A GREATER EFFICACY THAN DOCETAXEL Thomas Nelius*, Courtney Jarvis, Dalia Martinez-Marin, Stephanie Filleur, Lubbock, TX INTRODUCTION AND OBJECTIVES: Docetaxel and cabazitaxel have shown great promise in the treatment of metastatic Castration-Refractory Prostate Cancer (mCPRC) however, comparative studies are missing. Toxicities of these drugs are significant, urging the need to modify taxane-based regimens. Recently, low-dose metronomic (LDM) treatments using conventional chemotherapeutic drugs have shown benefits in CPRC in improving the effect of anti-angiogenic agents. Previously, we have demonstrated that LDM docetaxel in combination with PEDF curbs prostate tumor growth in vivo and that docetaxel/PEDF combination limits significantly the formation of metastases and prolongs survival in vivo. In the present study, we intended to compare the cytotoxic effect of cabazitaxel and docetaxel on CRPC cells in vitro and CL1 tumors in vivo. METHODS: Human PC3, DU145 cell lines were from ATCC. CL1 cells were obtained from LNCaP cells cultured under androgen deprivation. Tumor cell proliferation was assessed by crystal violet staining and cell cycle analyses. In vitro cytotoxicity assays were performed by co-culturing CL1 cells with RAW264.7 macrophages with PEDF and increasing concentrations of taxane drugs. For the in vivo studies, CL1 cells were genetically modified to stably express the DsRed Express protein þ/- PEDF. PEDF anti-tumor effects were assessed on established s.c. xenografts treated with docetaxel (5mg/kg ip every 4 day) as reference, cabazitaxel (5mg/kg ip every 4 days, 1mg/kg for 10 days, 0.5mg/kg q.a.d. and 0.1mg/kg daily) or placebo. RESULTS: Cabazitaxel limits in vitro cell proliferation with a greater efficacy than docetaxel in all CRPC cell lines tested. DU145
e817
presented the largest difference. High doses of taxane blocked tumor cells in mitosis, whereas LDM increased the SubG1 population. This effect was significantly higher only in DU145 cells treated with cabazitaxel. In vivo, 5mg/kg cabazitaxel delayed tumor growth more efficiently than 5mg/kg docetaxel. PEDF/5mg/kg cabazitaxel markedly delayed tumor growth compared to all treatments. Co-cultures demonstrated that tumor cells were engulfed with a higher frequency in combined treatments suggesting that inflammatory cells could be involved. CONCLUSIONS: Cabazitaxel has a greater efficacy compared to docetaxel both in vitro and in vivo. The data also reinforce PEDF as a promising anti-neoplasic agent in combination with LDM taxane chemotherapies. Source of Funding: The present study was performed with the financial support of the NIH/NCI grant R15CA161634.
MP66-05 PRETREATMENT SYSTEMIC INFLAMATORY RESPONSE PARAMETERS DO NOT PREDICT THE OUTCOME IN MEN WITH PROSTATE CANCER UNDERGOING RADICAL PROSTATECTOMY dric Poyet*, Jean-Pascal Adank, Etienne Keller, Ashkan Mortezavi, Ce Tenzin Rabgang, Bettina Pfister, Tullio Sulser, Thomas Hermanns, €rich, Switzerland Zu INTRODUCTION AND OBJECTIVES: Recent evidence indicates that inflammation is essential for development and progression of cancer. Several markers of systemic inflammation such as C-reactive protein (CRP), neutrophil-to-lymphocyte ratio (NLR) or platelet-tolymphocyte-ratio (PLR) have been shown to be associated with worse prognosis in different solid tumors. In this study we analyzed whether pretreatment systemic inflammatory response parameters are associated with clinicopathologic features or oncological outcome in men with clinically localized prostate cancer (PCa) undergoing radical prostatectomy. METHODS: Men who underwent radical prostatectomy for localized PCa in a tertiary referral centre between 2008 and 2013 were retrospectively analyzed. Patients were included if pre-operative complete blood count (CBC) was available. NLR and PLR were calculated from the CBC results. Time-dependent ROC curves were used to determine the optimal cutoff point for predicting recurrence-free survival (RFS). Mann-Witney-U-Test was used to test the associations between CRP, NLR and PLR with clinicopathological variables. RFS was calculated using both Kaplan-Meier and uni- and multivariate Cox regression methods. RESULTS: Our cohort consisted of 399 men with a median age of 64 (range 41-78), and median preoperative PSA of 7.1 (0.6e81.4) ng/ mL. Final pathology revealed 287 (71.9%) organ-confined PCa (T2) and 112 (28.1%) non-organ-confined disease (T3-4). Gleason Score (GS) 6 was present in 48 (12%), GS 7 in 266 (66.7%) and GS 8 in 85 (21.3%) patients. Median follow-up was 23 month (0-65). Median NLR for all patients was 2.67 (95% CI 1.30-5.85), Median PLR 153.89 (95% CI 85.71-302.50), and median CRP 1.2 (95% CI 0-11). Higher NLR, PLR and CRP were not associated with worse clinico-pathological parameters. ROC curves didn’t reveal any significant associations between RFS and pretreatment markers of systemic inflammation. NLR (hazard ratio [HR] 1.061; confidence interval [95% CI] 0.844-1.334; p¼0.612) PLR (HR 1.002; 95% CI 0.997-1.007; p¼0.389) and CRP (HR 0.817; 95% CI 0.639-1.044; p¼0.106) were not able to predict RFS in univariate analysis. T3-4 was the strongest clinicopathologic predictor in multivariate analysis for RFS (HR 4.358; 95% CI 2.021-9.399; p < 0.001). CONCLUSIONS: Pretreatment indexes of systemic inflammation do not predict the outcome in men with localized PCa. Localized disease may not upregulate the systemic inflammatory response to a measurable degree. This seems to be different in more advanced and metastatic disease. Source of Funding: none
Vol. 193, No. 4S, Supplement, Monday, May 18, 2015
therapeutic response. The goal of this study was to evaluate and genetically characterize exosome derived RNA (exoRNA) isolated from blood of metastatic CRPC patients. METHODS: Whole blood samples from 18 consented clinically annotated mCRPC patients and 1 normal control were collected. Exosomes were isolated with ultracentrifugation and exoRNA extracted. Following library prep, paired-end sequencing was performed using Illumina Hi-Seq 2000. A bioinformatics pipeline was used for data prepossessing including alignment, duplicate removal, normalization and variant calling. Visualization and differential analyses were performed with SNP & Variation Suite v8.x. RESULTS: In exoRNA there is evidence of extensive chromosomal rearrangement resulting in a myriad of gene fusions and isoforms. Through preliminary analyses we identified 39 genes commonly expressed in the exosomes of these mCRPC patients. These include PDPK1, USP9X, MAGI2, HMGA2 and PTGFR all of which have been previously expressed in prostate cancer tissue. A diverse variety of lcnRNA, ncRNA and miRNA were also identified in circulating exosomes. Validation of these alterations and additional analyses evaluating translocations and splice variants, PCR validation, and/or direct tumor based nucleic acid assays is ongoing. CONCLUSIONS: These preliminary analyses of circulating exoRNA have identified gene expression signatures of several prostate cancer associated transcripts. Ultimately, the identification of PCa associated transcripts in plasma derived exosomes provides evidence that exosomes and exosomal cargo may serve as biomarker in CRPC patients. Exosomes and exoRNA may provide otherwise unattainable insight into tumor evolution and disease progression. Additional studies evaluating the clinical relevance and prognostic value of exosomal RNA will be critical for biomarker development. Source of Funding: Tulane Cancer Center
MP66-04 CABAZITAXEL INHIBITS THE PROLIFERATION OF HUMAN CASTRATION-REFRACTORY PROSTATE CANCER CELLS IN VITRO AND ENHANCES THE ANTI-TUMOR PROPERTIES OF THE ANGIO-INHIBITORY PIGMENT EPITHELIUM-DERIVED FACTOR IN VIVO WITH A GREATER EFFICACY THAN DOCETAXEL Thomas Nelius*, Courtney Jarvis, Dalia Martinez-Marin, Stephanie Filleur, Lubbock, TX INTRODUCTION AND OBJECTIVES: Docetaxel and cabazitaxel have shown great promise in the treatment of metastatic Castration-Refractory Prostate Cancer (mCPRC) however, comparative studies are missing. Toxicities of these drugs are significant, urging the need to modify taxane-based regimens. Recently, low-dose metronomic (LDM) treatments using conventional chemotherapeutic drugs have shown benefits in CPRC in improving the effect of anti-angiogenic agents. Previously, we have demonstrated that LDM docetaxel in combination with PEDF curbs prostate tumor growth in vivo and that docetaxel/PEDF combination limits significantly the formation of metastases and prolongs survival in vivo. In the present study, we intended to compare the cytotoxic effect of cabazitaxel and docetaxel on CRPC cells in vitro and CL1 tumors in vivo. METHODS: Human PC3, DU145 cell lines were from ATCC. CL1 cells were obtained from LNCaP cells cultured under androgen deprivation. Tumor cell proliferation was assessed by crystal violet staining and cell cycle analyses. In vitro cytotoxicity assays were performed by co-culturing CL1 cells with RAW264.7 macrophages with PEDF and increasing concentrations of taxane drugs. For the in vivo studies, CL1 cells were genetically modified to stably express the DsRed Express protein þ/- PEDF. PEDF anti-tumor effects were assessed on established s.c. xenografts treated with docetaxel (5mg/kg ip every 4 day) as reference, cabazitaxel (5mg/kg ip every 4 days, 1mg/kg for 10 days, 0.5mg/kg q.a.d. and 0.1mg/kg daily) or placebo. RESULTS: Cabazitaxel limits in vitro cell proliferation with a greater efficacy than docetaxel in all CRPC cell lines tested. DU145
e817
presented the largest difference. High doses of taxane blocked tumor cells in mitosis, whereas LDM increased the SubG1 population. This effect was significantly higher only in DU145 cells treated with cabazitaxel. In vivo, 5mg/kg cabazitaxel delayed tumor growth more efficiently than 5mg/kg docetaxel. PEDF/5mg/kg cabazitaxel markedly delayed tumor growth compared to all treatments. Co-cultures demonstrated that tumor cells were engulfed with a higher frequency in combined treatments suggesting that inflammatory cells could be involved. CONCLUSIONS: Cabazitaxel has a greater efficacy compared to docetaxel both in vitro and in vivo. The data also reinforce PEDF as a promising anti-neoplasic agent in combination with LDM taxane chemotherapies. Source of Funding: The present study was performed with the financial support of the NIH/NCI grant R15CA161634.
MP66-05 PRETREATMENT SYSTEMIC INFLAMATORY RESPONSE PARAMETERS DO NOT PREDICT THE OUTCOME IN MEN WITH PROSTATE CANCER UNDERGOING RADICAL PROSTATECTOMY dric Poyet*, Jean-Pascal Adank, Etienne Keller, Ashkan Mortezavi, Ce Tenzin Rabgang, Bettina Pfister, Tullio Sulser, Thomas Hermanns, €rich, Switzerland Zu INTRODUCTION AND OBJECTIVES: Recent evidence indicates that inflammation is essential for development and progression of cancer. Several markers of systemic inflammation such as C-reactive protein (CRP), neutrophil-to-lymphocyte ratio (NLR) or platelet-tolymphocyte-ratio (PLR) have been shown to be associated with worse prognosis in different solid tumors. In this study we analyzed whether pretreatment systemic inflammatory response parameters are associated with clinicopathologic features or oncological outcome in men with clinically localized prostate cancer (PCa) undergoing radical prostatectomy. METHODS: Men who underwent radical prostatectomy for localized PCa in a tertiary referral centre between 2008 and 2013 were retrospectively analyzed. Patients were included if pre-operative complete blood count (CBC) was available. NLR and PLR were calculated from the CBC results. Time-dependent ROC curves were used to determine the optimal cutoff point for predicting recurrence-free survival (RFS). Mann-Witney-U-Test was used to test the associations between CRP, NLR and PLR with clinicopathological variables. RFS was calculated using both Kaplan-Meier and uni- and multivariate Cox regression methods. RESULTS: Our cohort consisted of 399 men with a median age of 64 (range 41-78), and median preoperative PSA of 7.1 (0.6e81.4) ng/ mL. Final pathology revealed 287 (71.9%) organ-confined PCa (T2) and 112 (28.1%) non-organ-confined disease (T3-4). Gleason Score (GS) 6 was present in 48 (12%), GS 7 in 266 (66.7%) and GS 8 in 85 (21.3%) patients. Median follow-up was 23 month (0-65). Median NLR for all patients was 2.67 (95% CI 1.30-5.85), Median PLR 153.89 (95% CI 85.71-302.50), and median CRP 1.2 (95% CI 0-11). Higher NLR, PLR and CRP were not associated with worse clinico-pathological parameters. ROC curves didn’t reveal any significant associations between RFS and pretreatment markers of systemic inflammation. NLR (hazard ratio [HR] 1.061; confidence interval [95% CI] 0.844-1.334; p¼0.612) PLR (HR 1.002; 95% CI 0.997-1.007; p¼0.389) and CRP (HR 0.817; 95% CI 0.639-1.044; p¼0.106) were not able to predict RFS in univariate analysis. T3-4 was the strongest clinicopathologic predictor in multivariate analysis for RFS (HR 4.358; 95% CI 2.021-9.399; p < 0.001). CONCLUSIONS: Pretreatment indexes of systemic inflammation do not predict the outcome in men with localized PCa. Localized disease may not upregulate the systemic inflammatory response to a measurable degree. This seems to be different in more advanced and metastatic disease. Source of Funding: none