Epidemiology, Diagnosis and Management of Penile Cancer: Results from the Spanish National Registry of Penile Cancer.

INTRODUCTION: Penile cancer (PC) is a rare malignancy with an overall incidence in Europe of 1/100,000 males/year. In Europe, few studies report the epidemiology, risk factors, clinical presentation, and treatment of PC. The aim of this study is to present an updated outlook on the aforementioned factors of PC in Spain. MATERIALS AND METHODS: A multicentric, retrospective, observational epidemiological study was designed, and patients with a new diagnosis of PC in 2015 were included. Patients were anonymously identified from the Register of Specialized Care Activity of the Ministry of Health of Spain. All Spanish hospitals recruiting patients in 2015 were invited to participate in the present study. We have followed a descriptive narration of the observed data. Continuous and categorical data were reported by median (p25th-p75th range) and absolute and relative frequencies, respectively. The incidence map shows differences between Spanish regions. RESULTS: The incidence of PC in Spain in 2015 was 2.55/100,000 males per year. A total of 586 patients were identified, and 228 patients from 61 hospitals were included in the analysis. A total of 54/61 (88.5%) centers reported ≤ 5 new cases. The patients accessed the urologist for visually-assessed penile lesions (60.5%), mainly localized in the glans (63.6%). Local hygiene, smoking habits, sexual habits, HPV exposure, and history of penile lesions were reported in 48.2%, 59.6%, 25%, 13.2%, and 69.7%. HPV-positive lesions were 18.1% (28.6% HPV-16). The majority of PC was squamous carcinoma (95.2%). PC was ≥cT2 in 45.2% (103/228) cases. At final pathology, PC was ≥pT2 in 51% of patients and ≥pN1 in 17% of cases. The most common local treatment was partial penectomy (46.9% cases). A total of 47/55 (85.5%) inguinal lymphadenectomies were open. Patients with ≥pN1 disease were treated with chemotherapy in 12/39 (40.8%) of cases. CONCLUSIONS: PC incidence is relatively high in Spain compared to other European countries. The risk factors for PC are usually misreported. The diagnosis and management of PC are suboptimal, encouraging the identification of referral centers for PC management.

[Living-donor renal transplant. State of the art.] / Trasplante renal de donante vivo. Estado actual.

Living donor kidney transplantation is the best therapeutic option in a patient with end-stage renal failure, because it provides excellent functionality and graft survival. Laparoscopic living donor nephrectomyis the gold-standard for obtaining the graft. In exper thands, different minimally invasive surgeries can be offered with the main advantage of improving the a esthetic results. Although there may be controversy regarding laparoscopic devices for vascular ligation during living donor nephrectomy, both endostaplers and locking clips have proven to be safe as long as the proper techniqueis performed. Living donor nephrectomy has minimal morbidity and mortality. Age and glomerular filtration rate of the donor candidate are prognostic factor of long-term renal failure. In relation to the implant surgery,robotic kidney transplantation is now probably at the beginning of its development. Published series still do not allow to clearly establish its role compared to conventional open surgery.

El trasplante renal de donante vivo suponela mejor opción terapéutica en un paciente con insuficiencia renal terminal, por su excelente funcionalidad y supervivencia del injerto. La nefrectomía laparoscópica de donante vivo es la técnica de elección para la obtención del injerto. En manos expertas, distintas variantes mínimamente invasivas pueden ofrecerse con la principal ventaja de mejorar los resultados estéticos del donante. Aunque pueda existir controversia en relación a los dispositivos laparoscópicos para la ligadura vascular durante la nefrectomía de donante vivo, tanto las endograpadoras como los clips con cierre tipo Hem-olokhan demostrado ser seguros siempre que se respete la técnica adecuada en su empleo. La nefrectomía de donante vivo no está exenta de una mínima morbi-mortalidad.La edad y el filtrado glomerular del candidato a donante son orientativas del riesgo de enfermedad renal a largo plazo. En relación a la cirugía del implante, el trasplante renal robótico se encuentra en los albores de su desarrollo. Las series publicadas no permiten aún establecer claramente su papel frente a la cirugía abierta convencional.

Trasplante renal de donante vivo. Estado actual / Immunosuppressive drugs and protocols for transplantation

El trasplante renal de donante vivo suponela mejor opción terapéutica en un paciente con insuficiencia renal terminal, por su excelente funcionalidad ysupervivencia del injerto. La nefrectomía laparoscópica de donante vivo es la técnica de elección para laobtención del injerto. En manos expertas, distintas variantes mínimamente invasivas pueden ofrecerse con laprincipal ventaja de mejorar los resultados estéticos deldonante. Aunque pueda existir controversia en relacióna los dispositivos laparoscópicos para la ligadura vascular durante la nefrectomía de donante vivo, tanto lasendograpadoras como los clips con cierre tipo Hem-olok han demostrado ser seguros siempre que se respetela técnica adecuada en su empleo. La nefrectomía dedonante vivo no está exenta de una mínima morbi-mortalidad. La edad y el filtrado glomerular del candidato a donante son orientativas del riesgo de enfermedad renala largo plazo. En relación a la cirugía del implante, eltrasplante renal robótico se encuentra en los albores desu desarrollo. Las series publicadas no permiten aún establecer claramente su papel frente a la cirugía abiertaconvencional.(AU)

Living donor kidney transplantation is thebest therapeutic option in a patient with end-stage renalfailure, because it provides excellent functionality andgraft survival. Laparoscopic living donor nephrectomyis the gold-standard for obtaining the graft. In experthands, different minimally invasive surgeries can be offered with the main advantage of improving the aesthetic results. Although there may be controversy regardinglaparoscopic devices for vascular ligation during livingdonor nephrectomy, both endostaplers and locking clipshave proven to be safe as long as the proper techniqueis performed. Living donor nephrectomy has minimalmorbidity and mortality. Age and glomerular filtrationrate of the donor candidate are prognostic factor oflong-term renal failure. In relation to the implant surgery,robotic kidney transplantation is now probably at thebeginning of its development. Published series still donot allow to clearly establish its role compared to conventional open surgery.(AU)

Nighttime Kidney Transplant From Donor With Controlled Cardiac Death: Greater Functionality at the Cost of More Complications?

BACKGROUND: Kidney transplantation surgery from controlled cardiac death donor (Maastricht III) is frequently performed at night, without taking into account the accumulated fatigue that the surgical team may experience. The objective of the study is to assess whether surgical complications and the functionality of the graft in the short and long term are affected by the time of day in which kidney transplantation from controlled cardiac death donors is performed. METHODS: A retrospective observational study was carried out. Patient were classified according to the start of surgery, daytime hours (8:00 AM to 7:59 PM), and nighttime hours (8:00 PM to 7:59 AM). Baseline and intraoperative parameters, postoperative complications, and parameters related to graft functionality were analyzed. RESULTS: A total of 77 patients were included: 37 patients had kidney transplantations performed during the daytime (48.05%), and 40 patients had kidney transplantations performed at nighttime (51.95%). No statistically significant differences were found between the baseline characteristics of both groups except for sex (55.0% men in daytime vs 78.4% men in nighttime, P = .03) and time on pretransplant dialysis (33.1 months in daytime vs 13.8 months in nighttime, P = .008). The incidence of surgical complications and the functionality of the graft was similar in both groups; however, the surgical time was shorter in night transplants (163.2 minutes in daytime vs 136.5 minutes at nighttime, P = .0006) CONCLUSION: The performance of kidney transplants at night is not associated, either in the short or long term, with an increase in surgical complications or conditions leading to the deterioration in the functionality of the graft.

Infección por SARS-CoV-2: implicaciones para la salud sexual y reproductiva. Una declaración de posición de la Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA) / SARS-CoV-2 infection: implications for sexual and reproductive health. A position statement of the Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA)

OBJETIVO: El objetivo de esta revisión es resumir la evidencia disponible sobre los posibles efectos adversos del SARS-CoV-2 en el sistema reproductor masculino y proporcionar una declaración de posición oficial de la Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA). MÉTODOS: Se realizó una búsqueda exhaustiva en las bibliotecas Pubmed, Web of Science, Embase, Medline, Cochrane y MedRxiv. RESULTADOS: No se ha confirmado la orquitis como una posible complicación de la infección por SARS-CoV-2. Un estudio informó que el 19% de los hombres con COVID-19 presentaban molestias escrotales sugestivas de orquitis viral, que no se pudo confirmar. Es posible que el virus no infecte los testículos directamente, si no que desencadene una respuesta autoinmune secundaria y que cause una orquitis autoinmune. COVID-19 se ha asociado con anormalidades en la coagulación por lo que la orquitis podría ser el resultado de una vasculitis segmentaria. Los datos disponibles sobre la presencia del virus en semen son contradictorios. Sólo un estudio informó de la presencia de ARN en el 15,8% de enfermos de COVID-19. La presencia de ácido nucleico o antígeno en el semen no implica la existencia de virus con capacidad de replicación o infección. En hombres con COVID-19 se ha observado un incremento significativo de LH en suero y una drástica disminución de la ratio T/LH y FSH/LH, congruente con un hipogonadismo subclínico. CONCLUSIONES: Los datos disponibles y los hallazgos de los estudios recientes se basan en tamaños de muestra pequeños y proporcionan informaciones contradictorias. Existe la posibilidad teórica de que pueda producirse daño testicular y posterior infertilidad después de la infección por COVID-19, por lo que especialmente para aquellos hombres en edad reproductiva, se debe sugerir consulta y evaluación de la función gonadal y análisis de semen. En cuanto a la posibilidad de transmisión sexual, no hay evidencia suficiente para respaldar la necesidad de que las parejas asintomáticas eviten las relaciones sexuales para protegerse contra la transmisión del virus. Se necesita más investigación para comprender los impactos a largo plazo del SARS-CoV-2 en la función reproductiva masculina, incluidos sus posibles efectos sobre la fertilidad y la función endocrina testicular

OBJECTIVE: The main objective of this revision is to summarize the current existing evidence of the potential adverse effects of SARS-CoV-2 on the male reproductive system and provide the recommendations of the Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA) concerning the implications of COVID-19 infection in the management of male infertilty patients and testicular endocrine dysfunction. METHODS: A comprehensive systematic literature search of the databases of PubMed, Web of Science, Embase, Medline, Cochrane and MedRxiv, was carried out. RESULTS: The presence of orchitis as a potential complication of the infection by SARS-CoV-2 has not yet been confirmed. One study reported that 19% of males with COVID-19 infection had scrotal symptoms suggestive of viral orchitis which could not be confirmed. It is possible that the virus, rather than infecting the testes directly, may induce a secondary autoimmune response leading to autoimmune orchitis. COVID-19 has been associated with coagulation disorders and thus the orchitis could be the result of segmental vasculitis. Existing data concerning the presence of the virus in semen are contradictory. Only one study reported the presence of RNA in 15.8% of patients with COVID-19. However, the presence of nucleic acid or antigen in semen is not synonyms of viral replication capacity and infectivity. It has been reported an increase in serum levels of LH in males with COVID-19 and a significant reduction in the T/LH and FSH/LH ratios, consistent with subclinical hypogonadism. CONCLUSIONS: The findings of recent reports related to the potential effects of COVID-19 infection on the male reproductive system are based on poorly designed, small sample size studies that provide inconclusive, contradictory results. Since there still exists a theoretical possibility of testicular damage and male infertilty as a result of the infection by COVID-19, males of reproductive age should be evaluated for gonadal function and semen analysis. With regard to the sexual transmission of the virus, there is not sufficient evidence to recommend asymptomatic couples to abstein from having sex in order to protect themselves from being infected by the virus. Additional studies are needed to understand the long-term effects of SARS-CoV-2 on male reproductive function, including male fertility potential and endocrine testicular function

Multicenter Prospective Study of Grafting With Collagen Fleece TachoSil in Patients With Peyronie's Disease.

BACKGROUND: A xenograft consisting of equine collagen coated with human fibrinogen and thrombin (TachoSil; Baxter, CA) has recently been introduced in grafting procedures for Peyronie's disease (PD). AIM: To describe the results of a multicenter prospective registry on patients with PD undergoing plaque incision or and grafting (PIG) or plaque excision and grafting (PEG) with collagen fleece TachoSil, to evaluate the efficacy and safety of this procedure. METHODS: A prospective non-controlled multicenter study of patients with PD was performed between May 2016 and March 2018. Patients from 10 centers with stable PD for at least 3 months, difficulties in sexual intercourse, normal erectile function with or without pharmacological treatment, curvature >45°, and/or penile shortening and/or complex deformities were included. All patients underwent PIG/PEG with collagen fleece TachoSil. OUTCOMES: The main outcome measure of this study were penile curvature correction (intraoperative), penile shortening (intraoperative), erectile function with the 5-item version of the International Index of Erectile Function (IIEF-5) and the Erection Hardness Score, subjective patient outcomes with non-validated questionnaires, and complications. RESULTS: A total of 52 patients were enrolled in the study. The mean (SD) preoperative penile curvature was 72.8° (17.0). PIG was the preferred technique (80.8%). Intraoperatively, complete curvature correction was achieved in 92.3%, and no significant penile shortening was recorded in 80.8% of subjects. Subjective penile shortening was reported in 83.3% of patients at 6 months. No objective measurement of penile curvature and length was recorded during follow-up. No statistically significant difference from the baseline was found in IIEF-5 and Erection Hardness Score at 3 or at 6 months, although 16.7% of men experienced a worsening of IIEF-5 scores and 14.3% required de novo phosphodiesterase type 5 inhibitor use. 6 months after surgery, 78.5% of men were satisfied with intervention. Swelling and ecchymosis/hematoma were the most common perioperative complications (40.4%). 2 cases (3.8%) of wound infection were recorded. At 6 months, 35.7% of patients reported mild penile hypesthesia. CLINICAL IMPLICATIONS: Our results confirm the high success rate of grafting with TachoSil, and the surgeon perceived low percentage of penile shortening. STRENGTH & LIMITATIONS: This is the first multicentre study on patients with PD undergoing grafting with TachoSil without concomitant placement of penile prosthesis. The main limitations are the short follow-up and the relatively small sample size. CONCLUSION: Grafting with TachoSil after PIG/PEG in patients with PD is an effective and safe procedure. Among the main advantages of this technique, there are ease of use of the graft and reduced operative time. Fernández-Pascual E, Manfredi C, Torremadé J, et al. Multicenter Prospective Study of Grafting With Collagen Fleece TachoSil in Patients With Peyronie's Disease. J Sex Med 2020;17:2279-2286.

[SARS-CoV-2 infection: implications for sexual and reproductive health. A position statement of the Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA)]. / Infección por SARS-CoV-2: implicaciones para la salud sexual y reproductiva. Una declaración de posición de la Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA).

OBJECTIVE: The main objective of this revision is to summarize the current existing evidence of the potential adverse effects of SARS-CoV-2 on the male reproductive system and provide the recommendations of the Asociación Española de Andrología, Medicina Sexual y Reproductiva (ASESA) concerning the implications of COVID-19 infection in the management of male infertilty patients and testicular endocrine dysfunction. METHODS: A comprehensive systematic literature search of the databases of PubMed, Web of Science, Embase, Medline, Cochrane and MedRxiv, was carried out. RESULTS: The presence of orchitis as a potential complication of the infection by SARS-CoV-2 has not yet been confirmed. One study reported that 19% of males with COVID-19 infection had scrotal symptoms suggestive of viral orchitis which could not be confirmed. It is possible that the virus, rather than infecting the testes directly, may induce a secondary autoimmune response leading to autoimmune orchitis. COVID-19 has been associated with coagulation disorders and thus the orchitis could be the result of segmental vasculitis. Existing data concerning the presence of the virus in semen are contradictory. Only one study reported the presence of RNA in 15.8% of patients with COVID-19. However, the presence of nucleic acid or antigen in semen is not synonyms of viral replication capacity and infectivity. It has been reported an increase in serum levels of LH in males with COVID-19 and a significant reduction in the T/LH and FSH/LH ratios, consistent with subclinical hypogonadism. CONCLUSIONS: The findings of recent reports related to the potential effects of COVID-19 infection on the male reproductive system are based on poorly designed, small sample size studies that provide inconclusive, contradictory results. Since there still exists a theoretical possibility of testicular damage and male infertilty as a result of the infection by COVID-19, males of reproductive age should be evaluated for gonadal function and semen analysis. With regard to the sexual transmission of the virus, there is not sufficient evidence to recommend asymptomatic couples to abstein from having sex in order to protect themselves from being infected by the virus. Additional studies are needed to understand the long-term effects of SARS-CoV-2 on male reproductive function, including male fertility potential and endocrine testicular function.

[Andrology and penile cancer. Recommendations during COVID-19 pandemia.] / Andrología y cáncer de pene. Recomendaciones durante la pandemia COVID-19.

PURPOSE: The COVID-19 pandemic which has affected Spain since the beginning of 2020 compels us to determine recomendations for the practice of Andrology in present times. MATERIALS AND METHODS: A web search is carried out in English and Spanish and a joint proposal is defined by experts in Andrology from different regions of Spain. RESULTS: Most diagnostic and therapeutic procedures in Andrology can be safey postponed during the COVID-19 pandemic. Online consultations and outpatient surgeries must be encouraged. Andrologic emergencies and penile cancer management should be considered high priority, and should be diagnosed and treated promptly even in the most severe phases of the pandemic.

INTRODUCCIÓN: La pandemia COVID-19 que ha afectado a España desde comienzos de 2020 obliga a definir unas recomendaciones para la práctica de la Andrología en la actualidad.MATERIAL Y MÉTODOS: Se realiza una búsqueda web en inglés y español y se define una propuesta conjunta por parte de expertos en Andrología de distintas regiones de España.RESULTADOS: La mayor parte de los procedimientos diagnósticos y terapéuticos en Andrología pueden ser demorados con seguridad durante la pandemia COVID-19. Se debe fomentar la consulta telemática y la cirugía ambulatoria. Las urgencias andrológicas y el manejo del cáncer de pene deben considerarse una prioridad alta, diagnosticándose y tratándose con brevedadi ncluso en las fases más severas de la pandemia.

Andrología y cáncer de pene. Recomendaciones durante la pandemia COVID-19 / Andrology and penile cancer. Recommendations during COVID-19 pandemia

INTRODUCCIÓN: La pandemia COVID-19 que ha afectado a España desde comienzos de 2020 obliga a definir unas recomendaciones para la práctica de la Andrología en la actualidad. MATERIAL Y MÉTODOS: Se realiza una búsqueda web en inglés y español y se define una propuesta conjunta por parte de expertos en Andrología de distintas regiones de España. RESULTADOS: La mayor parte de los procedimientos diagnósticos y terapéuticos en Andrología pueden ser demorados con seguridad durante la pandemia COVID-19. Se debe fomentar la consulta telemática y la cirugía ambulatoria. Las urgencias andrológicas y el manejo del cáncer de pene deben considerarse una prioridad alta, diagnosticándose y tratándose con brevedadi ncluso en las fases más severas de la pandemia

PURPOSE: The COVID-19 pandemic which has affected Spain since the beginning of 2020 compels us to determine recomendations for the practice of Andrology in present times. MATERIALS AND METHODS: A web search is carried out in English and Spanish and a joint proposal is defined by experts in Andrology from different regions of Spain. RESULTS: Most diagnostic and therapeutic procedures in Andrology can be safey postponed during the COVID-19 pandemic. Online consultations and outpatient surgeries must be encouraged. Andrologic emergencies and penile cancer management should be considered high priority, and should be diagnosed and treated promptly even in the most severe phases of the pandemic

Functional results in the treatment of localized prostate cancer. An updated literature review / Resultados funcionales del tratamiento del cáncer de próstata localizado. Revisión actualizada de la bibliografía

Prostate cancer is the second most frequently diagnosed cancer in men. The initial diagnosis is made in increasingly younger patients, so it seems to be essential to guarantee optimal functional results. We carried out a systematic search to define the functional results of each of the therapeutic options for localized prostate cancer. Radical prostatectomy generates a greater negative effect on urinary continence and erectile function compared to active surveillance and radiotherapy. Robotic surgery seems to offer better functional results, especially at the level of erectile function. Urinary and bowel symptoms are more pronounced after radiotherapy compared to other options. Patients must be warned of the possible functional results prior to choice of treatment

El cáncer de próstata es el segundo cáncer que se diagnostica con mayor frecuencia en varones. El diagnóstico inicial se establece en pacientes cada vez más jóvenes, por lo que parece que es fundamental para garantizar resultados funcionales óptimos. Se realizó una búsqueda sistemática para definir los resultados funcionales de cada una de las opciones terapéuticas para el cáncer de próstata localizado. La prostatectomía radical genera mayor efecto negativo sobre la continencia urinaria y la función eréctil en comparación con la vigilancia activa y la radioterapia. Parece que la cirugía robótica ofrece mejores resultados funcionales, sobre todo respecto a la función eréctil. Los síntomas urinarios e intestinales son más pronunciados después de la radioterapia en comparación con otras opciones. Se debe advertir a los pacientes de los posibles resultados funcionales antes de elegir el tratamiento

Functional results in the treatment of localized prostate cancer. An updated literature review.

Prostate cancer is the second most frequently diagnosed cancer in men. The initial diagnosis is made in increasingly younger patients, so it seems to be essential to guarantee optimal functional results. We carried out a systematic search to define the functional results of each of the therapeutic options for localized prostate cancer. Radical prostatectomy generates a greater negative effect on urinary continence and erectile function compared to active surveillance and radiotherapy. Robotic surgery seems to offer better functional results, especially at the level of erectile function. Urinary and bowel symptoms are more pronounced after radiotherapy compared to other options. Patients must be warned of the possible functional results prior to choice of treatment.

Methylation of tumor suppressor genes in a novel panel predicts clinical outcome in paraffin-embedded bladder tumors.

DNA methylation of tumor suppressor genes (TSGs) represents a frequent and early epigenetic event with potential applications for cancer detection and disease evolution. Our aim was to examine the stratification and prognostic biomarker role of the methylation of a novel panel of TSGs in bladder cancer. The methylation status of 18 TSGs was evaluated in bladder cancer cells (n=14) and paraffin-embedded primary bladder tumors (n=61), using a methylation-specific multiplex ligation-dependent probe amplification assay (MS-MLPA). Recurrence, progression, and disease-specific survival were analyzed using univariate and multivariate Cox models. PRDM2, HLTF, ID4, DLC1, BNIP3, H2AFX, CACNA1G, TGIF, and CACNA1A were discovered methylated in bladder cancer. The methylation of RUNX3 (p=0.026), TWIST1 (p=0.009), SFRP4 (p=0.002), and CCND2 (p=0.027) correlated to tumor stage. Univariate analyses indicated prognostic associations for recurrence (DLC1, SFRP5, H2AFX, CACNA1G), progression (DLC1, SFRP5, CACNA1G), disease-specific (PRDM2, DLC1, SFRP5, CACNA1G, and TIMP3), and overall survival (SFRP5 and TIMP3). In multivariate analyses, several TSGs remained as independent prognosticators for recurrence (SFRP5, H2AFX), progression (CACNA1G), and disease-specific survival (SFRP5). Thus, a novel set of TSGs was identified, frequently methylated in bladder cancer cells and tumors. TSG methylation allowed histopathologic and outcome stratification using paraffin-embedded tumors. This is clinically relevant by offering a strategy for the management of patients affected with uroepithelial neoplasias in pathology routine laboratories.

Methylation of a novel panel of tumor suppressor genes in urine moves forward noninvasive diagnosis and prognosis of bladder cancer: a 2-center prospective study.

PURPOSE: Changes in DNA methylation of tumor suppressor genes early in carcinogenesis represent potential indicators of cancer detection and disease evolution. We examined the diagnostic, stratification and prognostic biomarker roles in urine of the methylation of a novel panel of tumor suppressor genes in bladder cancer. MATERIAL AND METHODS: We evaluated the methylation of 18 tumor suppressor genes in 2 prospective, independent sets of urine samples (training set of 120 preparations and validation set of 128) from patients with bladder cancer (170) and controls (78) using methylation specific multiplex ligation-dependent probe amplification. Diagnostic performance was evaluated with ROC curves. Recurrence, progression and disease specific survival were analyzed using univariate and multivariate Cox models. RESULTS: PRDM2, HLTF, ID4, DLC1, BNIP3, H2AFX, CACNA1G, TGIF and CACNA1A were methylated in bladder cancer. CCND2, SCGB3A1, BNIP3, ID4 and RUNX3 were the most frequently methylated tumor suppressor genes in each urine set. Methylation of several tumor suppressor genes correlated with clinicopathological variables, such as stage, tumor grade, focality or age. ROC analysis revealed significant diagnostic accuracy for RUNX3 and CACNA1A in the training set, and for RUNX3 and ID4 in the validation set. On univariate and multivariate analysis CACNA1A methylation correlated with recurrence in the training set, while in the validation set PRDM2 and BNIP3 were significantly associated with recurrence and disease specific survival, respectively. CONCLUSIONS: Tumor suppressor gene methylation allowed for histopathological and clinical stratification. Urine methylation has noninvasive usefulness not only for diagnostic assessment but also as independent bladder cancer prognosticators.

miR-143, miR-222, and miR-452 are useful as tumor stratification and noninvasive diagnostic biomarkers for bladder cancer.

Altered microRNA (miRNA) expression may occur early in bladder cancer and may play a role in carcinogenesis and tumor behavior. We evaluated whether alterations in miRNA expression could improve disease stratification and outcome prognosis in bladder tumors and noninvasive diagnosis in urinary samples. miR-143, miR-222, and miR-452 expression levels were analyzed by quantitative RT-PCR (RT-qPCR) in paired urinary and matching tumors and in two independent prospective series of tumors and urinary specimens. Differential expression of miR-143, miR-222, and miR-452 in urine were verified by in situ hybridization in matching tumors. Tumor miRNA expression by RT-qPCR correlated with tumor grade, size, and presence of carcinoma in situ for miR-222, recurrence (miR-222 and miR-143), progression (miR-222 and miR-143), disease-specific survival (miR-222), and overall survival (miR-222). Protein expression patterns of potential miRNA targets, including vascular endothelial growth factor, BCL2, v-erb-b2 erythroblastic leukemia viral oncogene (ERBB) homolog 3, and ERBB4, were evaluated by IHC in tissue arrays containing tumors for which miRNAs were assessed by RT-qPCR. Target expression correlated with expression of their predicted regulatory miRNAs, recurrence (ERBB3), progression (ERBB4), disease-specific survival (ERBB3 and ERBB4), and overall survival (ERBB3 and ERBB4). Furthermore, RT-qPCR of miR-452 (area under the curve, 0.848) and miR-222 (area under the curve, 0.718) in urine provided high accuracies for bladder cancer diagnosis. Thus, bladder tumors were characterized by changes in miRNA expression that could aid in tumor stratification and clinical outcome prognosis, and miRNAs were detected in urinary specimens for noninvasive diagnosis.