Safwan Escaf

Myopodin Methylation is Associated With Clinical Outcome in Patients With T1G3 Bladder Cancer

PURPOSE Bacillus Calmette-Guerin is standard treatment to decrease tumor recurrence and delay progression of high risk, nonmuscle invasive bladder tumors. However, it is not yet clear which T1G3 cases are more prone to more aggressive clinical behavior or susceptible to respond to bacillus Calmette-Guerin. We evaluated the role of myopodin methylation as a clinical outcome prognosticator and predictive biomarker for the bacillus Calmette-Guerin response in patients with T1G3 bladder tumors. MATERIALS AND METHODS We analyzed the methylation status of myopodin in tumor specimens from 170 patients with T1G3 bladder cancer, including a subset of 108 who underwent bacillus Calmette-Guerin treatment. Myopodin methylation was assessed by methylation specific polymerase chain reactions. Recurrence, progression to muscle invasive tumors and disease specific overall survival were analyzed using competing risks regression analysis. RESULTS Of the 170 cases analyzed 72 recurred (42.4%) and 36 progressed (21.2%). A total of 24 patients (14.1%) died of the disease. Univariate and multivariate survival analysis revealed that myopodin methylation was significantly associated with an increased recurrence rate (p = 0.004), progression (p = 0.002) and shorter disease specific overall survival (p = 0.020). In a subset treated with bacillus Calmette-Guerin myopodin methylation was also related to an increased recurrence rate (p = 0.011), progression (p = 0.030) and shorter disease specific overall survival (p = 0.028). CONCLUSIONS Epigenetic analysis revealed that myopodin methylation was associated with tumor aggressiveness and clinical outcome in patients with T1G3 disease. Myopodin methylation distinguished patients responding to bacillus Calmette-Guerin from those who may require a more aggressive therapeutic approach.

Molecular Classification of Non–Muscle-Invasive Bladder Cancer (pTa Low-Grade, pT1 Low-Grade, and pT1 High-Grade Subgroups) Using Methylation of Tumor-Suppressor Genes

The role of epigenetics in distinguishing pathological and clinical subgroups in bladder cancer is not fully characterized. We evaluated whether methylation of tumor-suppressor genes (TSGs) would classify non-muscle-invasive (NMI) bladder cancer subgroups and predict outcome. A retrospective design included the following paraffin-embedded primary NMI tumor types (n = 251): pTa low grade (LG) (n = 79), pT1LG (n = 81), and pT1 high grade (HG) (n = 91). Methylation of 25 TSGs was measured using methylation-specific, multiplex, ligation-dependent probe amplification. The TSGs most frequently methylated in the overall series were STK11 (96.8%), MGMT2 (64.5%), RARB (63.0%), and GATA5 (63.0%). TSG methylation correlated to clinicopathological variables in each subgroup and in the overall NMI series. Methylation of RARB, CD44, PAX5A, GSTP1, IGSF4 (CADM1), PYCARD, CDH13, TP53, and GATA5 classified pTa versus pT1 tumors whereas RARB, CD44, GSTP1, IGSF4, CHFR, PYCARD, TP53, STK11, and GATA5 distinguished LG versus HG tumors. Multivariate analyses indicated that PAX5A, WT1, and BRCA1 methylation independently predicted recurrence in pTaLG, PAX6, ATM, CHFR, and RB1 in pT1LG disease; PYCARD, in pT1HG disease; and PAX5A and RB1, in the overall series. Methylation of TSGs provided a molecular classification of NMI disease according to clinicopathological factors. Furthermore, TSG methylation predicted recurrence in NMI subgroups.

Polyamine-modulated Factor-1 Methylation Predicts Bacillus Calmette-Guérin Response in Patients with High-grade Non–muscle-invasive Bladder Carcinoma

BACKGROUND Bacillus Calmette-Guérin (BCG) is a standard treatment to reduce tumor recurrence and delay progression of high-risk non-muscle-invasive (NMI) bladder tumors. However, it is not clear yet which patients are more likely to respond to BCG. OBJECTIVE The aim was to evaluate the role of polyamine-modulated factor-1 (PMF-1) methylation in predicting clinical outcome of T1 high-grade (T1HG) bladder tumors treated with BCG. DESIGN, SETTING, AND PARTICIPANTS In a retrospective design, PMF-1 methylation was analyzed on tumor specimens belonging to 108 patients with T1HG NMI bladder cancer undergoing BCG treatment. Median follow-up was 77 mo (range: 5-235 mo). OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS PMF-1 methylation was assessed by methylation-specific polymerase chain reactions. Recurrence, progression into muscle-invasive tumors, and disease-specific survival rates were analyzed using competing risks regression analysis. RESULTS AND LIMITATIONS Among the 108 patients analyzed, 35 had recurring disease (32.4%), 21 progressed (19.4%), and 16 died of disease (14.8%); 71.3% of tumors had PMF-1 methylation. Univariate analyses using cumulative incidence curves revealed that an unmethylated PMF-1 was significantly associated with increased recurrence (p=0.026), progression (p=0.01), and shorter disease-specific survival (log-rank, p=0.03). Multivariate analyses indicated that among sex, age, focality, tumor size, and concomitant carcinoma in situ, only PMF-1 methylation provided significant hazard ratios (HRs) for recurrence of (HR: 2.032; p=0.042), and progression (HR: 2.910; p=0.020). Limitations of the study include its retrospective design, lymphovascular invasion status not available, short maintenance BCG, and that a second transurethral resection was not performed. CONCLUSIONS Epigenetic analyses revealed that the methylation status of PMF-1 was associated with the clinical outcome of patients with T1HG tumors undergoing BCG treatment. An unmethylated PMF-1 correlated to recurrence and progression in T1HG disease using univariate and multivariate analyses. Thus, assessing the methylation status of PMF-1 may serve to distinguish patients responding to BCG from those who may require more aggressive therapeutic approaches.

Sympathetic skin response in patients with erectile dysfunction

To evaluate the role of the sympathetic skin response (SSR) in men with erectile dysfunction (ED), focusing on detecting SSR in the penis.

Stromal factors involved in human prostate cancer development, progression and castration resistance

PurposeTo detect new predictive markers from the prostate cancer tissue, to study the expression by cultured cancer-associated fibroblasts (CAFs) of stromal factors implicated in prostate carcinogenesis, and to compare their expressions in localized, metastatic, castration-sensitive (CSCP), castration-resistant prostate tumors (CRCP) as well as in fibroblasts from benign prostatic hyperplasia (BPH).Materials and methodsThe genomic expression of 20 stroma-derived factors, including the androgen receptor (AR), growth factors (FGF2, FGF7, FGF10, HGF, TGFβ, PDGFB), protein implicated in invasion (MMP-2, MMP-9 and MMP-11), inflammation (IL-6, IL-17, STAT-3 and NFκB), stroma/epithelium interaction (CDH11, FAP, CXCL12 and CXCL14) and chaperones (HPA1A and HSF1), was evaluated in cultured fibroblasts both from BHP and prostate carcinomas (PCa). After isolation and culture of fibroblasts by biopsy specimens, RNA was isolated and genomic studies performed.ResultsFinally, 5 BPH and 37 PCa specimens were selected: clinically localized (19), metastatic (5), CSCP (7) and CRPC (6). Interleukin-17 receptor (IL-17RB) was highly expressed in CAFs compared with fibroblasts from BPH. However, metalloproteinase-2 and chemokine ligand 14 (CXCL14) were expressed at higher levels by fibroblasts from BPH. The fibroblastic growth factor-7 was highly expressed by CAFs from localized tumors, but metalloproteinase-11 in metastatic tumors. MMP-11, androgen receptor (AR) and heat–shock-70kda-protein-1A (HSPA1A) expressions were significantly higher in CAFs from CRPC.ConclusionsThese results demonstrate a CAFs heterogeneity among prostate carcinomas with regard to some molecular profile expressions that may be relevant in tumor development (IL-17RB), progression (MMP-11) and castration resistance (AR, MMP-11 and HSPA1A).

Etiología neurógena en pacientes con disfunción eréctil.

OBJETIVOS La disfuncion erectil (DE) es una alteracion cuya prevalencia es elevada y aumenta con la edad. Se estima que en Espana afecta al 18,9% de los varones de 25 a 70 anos. En la mayor parte de los casos es de origen multifactorial y en su patogenia se admite la influencia de enfermedades sistemicas, farmacos de diferentes tipos, factores psicogenos, patologias cardiovasculares, endocrinopatias y alteraciones neurologicas. La disfuncion erectil de causa neurologica puede tener su origen a nivel del Sistema Nervioso Central o Periferico. Entre las posibles causas de disfuncion erectil neurogena de origen central estarian tumores, accidentes cerebrovasculares, encefalitis, Enfermedad de Parkinson, Esclerosis Multiple y otras enfermedades desmielinizantes, demencias, degeneracion olivopontocerebelosa y epilepsia. Las mielopatias de cualquier etiologia, pueden ser dependiendo de su localizacion o extension, causas de disfuncion erectil.A nivel periferico pueden ser causa de DE las alteraciones de las vias sensitivas que constituyen el brazo aferente del reflejo espinal de la ereccion y las de las vias eferentes vegetativas o somaticas que median en la vasodilatacion arterial, la relajacion del musculo liso cavernoso o la contraccion de la musculatura estriada del suelo de la pelvis. La finalidad de este trabajo es revisar detalladamente las causas mas relevantes de DE de origen neurogeno, sus mecanismos etiopatogenicos y los abordajes terapeuticos que en la actualidad se consideran mas adecuados para cada caso particular. CONCLUSIONES La correcta aproximacion diagnostica al paciente con DE pasa por identificar, en la medida de lo posible, los factores etiopatogenicos implicados su origen. En este sentido, la deteccion e identificacion, de la posible presencia del factor de riesgo neurogeno, contibuira a un mejor entendimiento de sus mecanismos fisiopatologicos y con ello a una aproximacion diagnostica, pronostica y terapeutica mas adecuada especialmete en aquellos pacientes refractarios a la terapia de primera linea.

Analysis of the expression of interleukins, interferon β, and nuclear factor-κ B in prostate cancer and their relationship with biochemical recurrence.

There are accumulating epidemiological, experimental, and genetic data supporting that prostate inflammation may contribute to prostate carcinogenesis, and several inflammatory-related molecules have been linked to tumorigenesis and prognosis in several tumors. The aim of this study was to evaluate tumor expression of inflammatory-related factors in prostate carcinomas and their possible relationship with biochemical recurrence (elevation of prostate-specific antigen serum levels). An immunohistochemical study was conducted using tissue microarrays and specific antibodies against interleukin-1&bgr; (IL-1&bgr;), IL-6, IL-10, IL-17, interferon &bgr; (IFN&bgr;), and nuclear factor-&kgr; B (NF-&kgr;B). Determinations in cancer specimens from 118 patients with primary prostate cancer (78 without and 40 with recurrence during the follow-up period) were performed. Immunostaining for all the studied proteins was localized both in tumor cells and in stromal cells in the majority of tumors. High-score values for IL-1&bgr; or low-score values for IFN&bgr; were significantly associated with biochemical recurrence. The analysis defined a score value of 160 for IL-1&bgr; and of 170 for IFN&bgr; as the optimal cutoff points that identified 32.7% and 73.2% of patients, respectively, having high probability of biochemical recurrence. Multivariate analysis according to a Cox model indicated that the cutoff point 170 for IFN&bgr; (P=0.035) was an independent factor associated with biochemical recurrence in patients with prostate cancer. Both IL-1&bgr; and IFN&bgr; may be new biomarkers to distinguish high-risk/low-risk patients with prostate cancer, and to select appropriate therapeutic approaches.

Relationship between metalloprotease expression in tumour and stromal cells and aggressive behaviour in prostate carcinoma: Simultaneous high-throughput study of multiple metalloproteases and their inhibitors using tissue array analysis of radical prostatectomy samples

Abstract Objective. The aim of this study was to detect a potential association between clinicopathological factors of prostate cancer aggressiveness and the expression of matrix metalloproteases and their inhibitors in tumour and stromal cells. Material and methods. A tissue array technique and immunochemistry with specific antibodies against matrix metalloproteinases (MMPs)-1, 2, 7, 9, 11, 13, 14, and their tissue inhibitors (TIMPs)-1, 2 and 3 were used to analyse the surgical specimens of 133 patients treated by radical prostatectomy. For each antibody preparation, the cellular location of immunoreactivity was determined. Results. The expression of MMP-2 was negatively associated with high tumour grade. With regard to stromal fibroblasts, TIMP-3 expression was positively associated with histological grade. MMP-7 expression was negatively associated with pretreatment serum levels of PSA, whereas MMP-13 was positively associated with higher levels of the antigen. TIMP-2 expression by mononuclear inflammatory cells correlated significantly and negatively with tumour grade. Conclusions. The expression of TIMP-3 by fibroblasts was associated with a higher Gleason score. An increased expression of MMP-13 by fibroblasts was associated with a greater preoperative level of PSA. In contrast, MMP-2 expression by tumour as well as TIMP-2 expression by peritumoral inflammatory cells was associated with less aggressive prostate carcinoma characteristics.

Determination of a New Cut-Off Value for the Urinary BTAtrak to Optimize the Detection of Recurrences in the Follow-Up of Patients with Superficial Bladder Carcinoma

The present study aimed to prospectively reassess the cut-off value of the urinary BTAtrak assay in order to optimize the detection of high and middle risk recurrent tumors during the follow-up of superficial bladder cancer. Patients and Methods: We prospectively investigated sensitivity and specificity values of BTAtrak (cut-off: 8 U/ml), cytology, hemoglobin dipstick and their combinations in 956 patients monitored for bladder cancer recurrence. All of the patients (n = 2) with upper urinary tract tumours (UUTT) showed a positive BTAtrak and were not included in the study. We found 215 bladder neoplasms, 149 were papillary and the remaining 66 were solid tumors. There were 115 (53.5%) single tumors and 100 (46.5%) multiple tumors. The mean size of the tumors was 1.78 ± 1.8 cm with 80 (37.2%) minor than 0.5 cm, 103 (47.9%) from 0.5 to 3 cm and 32 (14.9%) tumors greater than 3 cm. Results: BTAtrak (cut-off: 8 U/ml) plus cytology demonstrated the highest overall sensitivity and detected all high-risk tumors (Ta-1G3/Tis and T2-4). Specificity increased considerably when radiotherapy, urinary tract infection (UTI), urethral catherization as well as intravesical instillations within 3 months before BTAtrak determination were not included. The combinations of cytology and BTAtrak resulted negative in 42.6% of cases with a BTAtrak cut-off of 8 U/ml. Conclusion: The combination of cytology and BTAtrak using a lower cut-off level (8 U/ml) demonstrated to be a very sensitive marker to detect most of dangerous tumors in the follow-up of superficial bladder cancer.

P28.4 Role of penile sympathetic skin response in the neurophysiological evaluation of erectile dysfunction

neurocontrol mechanism, but need to be reinforced, perhaps in correlation with the underline pathophysiology of symptom. In 5 pts (4.5%) implanted for dysfunctional voiding in whom SNM was switched off with a persistent clinical efficacy no difference in PSEPs at T1 and T2 was seen: if a physiological restoration is achieved with SNM, the effect on neurocontrol mechanism persists in a normal fashion. Conclusion: NPE shed light to the mechanism by which central nervous system modify its organization under SNM.