Unraveling the Metabolic Alterations Induced by Zika Infection in Prostate Epithelial (PNT1a) and Adenocarcinoma (PC-3) Cell Lines.

The outbreak of Zika virus infection in 2016 led to the identification of its presence in several types of biofluids, including semen. Later discoveries associated Zika infection with sexual transmission and persistent replication in cells of the male reproductive tract. Prostate epithelial and carcinoma cells are favorable to virus replication, with studies pointing to transcriptomics alterations of immune and inflammation genes upon persistence. However, metabolome alterations promoted by the Zika virus in prostate cells are unknown. Given its chronic effects and oncolytic potential, we aim to investigate the metabolic alterations induced by the Zika virus in prostate epithelial (PNT1a) and adenocarcinoma (PC-3) cells using an untargeted metabolomics approach and high-resolution mass spectrometry. PNT1a cells were viable up to 15 days post ZIKV infection, in contrast to its antiproliferative effect in the PC-3 cell lineage. Remarkable alterations in the PNT1a cell metabolism were observed upon infection, especially regarding glycerolipids, fatty acids, and acylcarnitines, which could be related to viral cellular resource exploitation, in addition to the over-time increase in oxidative stress metabolites associated with carcinogenesis. The upregulation of FA20:5 at 5 dpi in PC-3 cells corroborates the antiproliferative effect observed since this metabolite was previously reported to induce PC-3 cell death. Overall, Zika virus promotes extensive lipid alterations on both PNT1a and PC-3 cells, promoting different outcomes based on the cellular metabolic state.

Predictive Value of Neutrophil to Lymphocyte Ratio in the Diagnosis of Significant Prostate Cancer at Initial Biopsy: A Comparison with Free Percent Prostate Specific Antigen, Prostate Specific Antigen Density and Primary Circulating Prostate Cells.

INTRODUCTION: An elevated serum PSA is the only biomarker routinely used in screening for prostate cancer to indicate a prostate biopsy. However, it is not specific for prostate cancer and the neutrophil/lymphocyte ratio has been suggested as an alternative. We present a prospective study of men with an elevated PSA and compare the neutrophil/lymphocyte ratio, free percent PSA, PSA density and the presence of circulating prostate cells to detect clinically significant prostate cancer at first biopsy. PATIENTS AND METHODS: Prospective study of consecutive men with a PSA 4-10 ng/ml referred for initial prostate biopsy, the results were compared with the neutrophil/lymphocyte ratio, free percent PSA and PSA density. Circulating prostate cells (CPCs) were detected using immunocytochemistry. The blood sample was taken immediately before the prostate biopsy. RESULTS: 1,223 men participated, 38% (467) of whom had prostate cancer detected, of these 322 were clinically significant. The area under the curves were for neutrophil/lymphocyte ratio, free percent PSA, PSA density and CPC detection were 0.570, 0.785, 0,620 and 0.844 respectively. Sensitivity/specificity were 0.388/0.685, 0.419/0.897, 0.598/0.624 and 0.966/0.786 respectively. The neutrophil/lymphocyte ratio did not differentiate between benign and malignant disease. CONCLUSIONS: The neutrophil/lymphocyte ratio did not discriminate between benign and malignant prostatic disease in patients with a PSA between 4-10ng/ml.

The presence of secondary circulating prostate tumour cells determines the risk of biochemical relapse for patients with low- and intermediate-risk prostate cancer who are treated only with external radiotherapy.

INTRODUCTION: The classification of patients with prostate cancer is used to determine treatments based on risk factors. The presence of secondary circulating prostate tumour cells (CPCs) detected in peripheral blood after a curative treatment has been associated with a worse prognosis. We present a prospective study of CPC detection post radiotherapy and the oncological results. PATIENTS AND METHODS: All of the patients classified as low and intermediate risk that were treated with radiotherapy were included. Three months after finishing treatment, an 8-ml blood sample was taken to detect CPCs. Mononuclear cells were obtained using gel centrifugation, and CPCs were identified using immunocytochemistry with anti-prostate-specific antigen. Patients were classified as low-risk CPC positive or negative and intermediate-risk CPC positive or negative. The biochemical relapse-free survival analysis was determined based on a follow-up of up to 15 years using the Kaplan-Meier and Cox regression models. Biochemical failure was defined according to the Pheonix II criteria. RESULTS: Of 241 patients, 181 (75.1%) were classified as low risk and 60 (24.9%) as intermediate risk. Biochemical failure was observed in 27.1% (49/181) of the low-risk prostate cancer participants and in 53.3% (32/60) of intermediate-risk participants after 15 years of follow-up. 20.4% (37/181) of the low-risk cancer participants had detectable CPCs in comparison with 43.3% (26/60) of the intermediate-risk cancer participants (p < 0.001 overall risk 2.98, confidence interval (CI) 95% 1.59-5.56; relative risk 2.12, CI 95% 1.41-3.19). Positive CPC patients had a worse prognosis, and a shorter time period until biochemical relapse, regardless of risk group. The biochemical relapse-free survival curves show that intermediate-risk participants who were CPC negative had a higher survival rate and slower disease progression than those participants who were low risk but CPC positive. CONCLUSIONS: CPC detection is a risk factor for biochemical relapse and could be useful in identifying patients that will need additional treatment.

How localized is pathologically localized prostate cancer? The use of secondary circulating prostate cells as a marker of minimal residual disease and their association with patient outcome.

OBJECTIVE: To determine the prognostic value of secondary circulating prostate cells (CPCs) in men with pT2 prostate cancer treated with radical prostatectomy. MATERIAL AND METHODS: Prospective observational study was performed in men with pathologically confined prostate cancer who had been treated with radical prostatectomy. CPCs were obtained by differential gel centrifugation from 8 mL venous blood and identified by standard immunocytochemistry using anti-Prostate Specific Antigen (PSA) monoclonal antibody. A positive test was defined as ≥1 PSA staining cell/blood sample. Biochemical failure was defined as a serum PSA >0.2 ng/mL. Age, PSA at diagnosis, pT2a versus pT2b/c, Gleason score and the presence/absence of CPCs were compared with patient outcomes using Kaplan-Meier curves and Cox's hazard model. RESULTS: Hundred and ninety-one men participated in the study, 107 (44.0%) had pT2b/c disease, 25 (13.1%) had a Gleason score ≥7, and 39 (20.4%) were positive for CPCs. Biochemical failure occurred in 39 (20.4%) patients which was associated with a Gleason score ≥ 7 and CPCs (+). Survival rates at 3, 5 and 10 years for men with CPC (-) and CPC (+) were 100%, 100% and 89.6%, and 74.4%, 64.1% and 18.5% respectively (HR: 18.70). The median time to failure was 5.1 years in CPC (+) men versus 8.1 years in CPC (-) patients. CONCLUSION: Secondary CPC is a marker for minimal residual disease and it is associated with a worse prognosis. The lead time to failure over serum PSA is approximately 5 years. However they do not define whether the failure is local or systemic.

The presence of primary circulating prostate cells is associated with upgrading and upstaging in patients eligible for active surveillance.

Active surveillance (AS) is a considered treatment option for men with low or very low-risk prostate cancer. However, on repeat biopsy some 25% were upgraded and recommended for active treatment. We compare the presence or absence of primary circulating prostate cells (CPCs) with the clinical pathological findings after radical prostatectomy in men fulfilling the criteria for active surveillance and the risk of reclassification for active observation (AO). METHODS AND PATIENTS: A single centre observational study was done involving 102 men who fulfilled the Epstein criteria for AS and underwent radical prostatectomy as mono-therapy for prostate cancer. The patients were classified according to the presence or absence of CPCs detected immediately before the prostate biopsy. Mononuclear cells were obtained by differential gel centrifugation of 8 mL of venous blood and CPCs identified using immunocytochemistry with anti-PSA and anti-P504S. A positive CPC test was defined as at least 1 PSA (+), P504S (+) cell detected/blood sample. The surgical specimen was analysed for Gleason score and pathological stage. RESULTS: A total of 25 out of 102 (24.5%) men were upgraded based on the pathological findings of the surgical specimen. Among which 45 (44%) men were positive for CPCs. They were younger, 63.9 versus 68.1 years (p = 0.0148), had a lower frequency of pT2 or lower disease (64.4% versus 91.2% p <0.001), higher median Gleason scores (6 versus 5 p < 0.001) in both the biopsy and surgical specimens, and a higher frequency of upgrading 44% versus 9% (p < 0.001). CONCLUSIONS: In men fulfilling the criteria for AS, the presence of primary CPCs suggests a high risk for disease upgrade and therefore these men may not be ideal for observational therapy. Further studies with a larger population are warranted.

Células prostáticas circulantes para la detección de cáncer / Circulating prostate cells for cancer detection

Introducción: El antígeno prostático específico (APE) es el marcador en suero más utilizado para la patología prostática, y el único empleado para la detección del cáncer de próstata. Si bien el APE es sumamente específico para tejido prostático, un nivel elevado no lo es para cáncer de próstata, ya que se puede ver elevado también en la enfermedad prostática benigna. Existe la necesidad de un examen simple para definir la necesidad de una biopsia prostática en hombres con un APE alterado. El examen de detección de células prostáticas malignas en sangre (CPMs) puede ser un candidato para la detección precoz del cáncer de próstata (CP). Métodos y pacientes: Analizamos de manera prospectiva un grupo de pacientes que fueron sometidos a una biopsia inicial por sospecha de cáncer de próstata y también a una segunda biopsia en el seguimiento posterior. Los resultados de la biopsia fueron comparados con las células prostáticas malignas circulantes (CPMC), las cuales fueron identificadas en muestras de sangre tomadas antes de la realización de la biopsia utilizando inmunocitoquímica estándar. El objetivo fue determinar la capacidad diagnóstica de las CPMC antes de la primera, la segunda y la tercera biopsia prostática. Resultados: En total, 423 pacientes consecutivos participaron en el estudio, con una edad promedio de 65.3 ± 8.9 años y un APE (mediana) de 5.28 ng/ml (RIQ 4.36-7.94 ng/ml). De ellos, 138 (32.6%) tuvieron un cáncer detectado en la biopsia inicial. La prueba de la detección de CPCs tuvo una sensibilidad de 0.89 (IC 95%: 0.82 a 0.94) y una especificidad de 0.89 (IC 95%: 0.84 a 0.92). De los 423 pacientes, a 125 se les reallizó una segunda biopsia, y a 57, una tercera, las CPCs lograron una sensibilidad y especificidad de 0.89/0.87 y 0.88/0.96 en la segunda y tercera biopsia, respectivamente, con un valor predictivo negativo y positivo de 0,65 / 0,97 y 0,88 / 0,95 en la segunda y tercera biopsia, en ese orden. Conclusiones: Las CPCs utilizadas en conjunto con las pruebas de tamizaje actuales, APE y tacto rectal, pueden mejorar la efectividad del tamizaje, reduciendo la frecuencia de biopsias negativas, así como su número total y sus complicaciones. Además, el costo-beneficio para el sistema de salud público en términos de utilización de recursos es positivo

Introduction: The prostate-specific antigen (PSA) is the most frequently used serum marker for the detection of prostatic disease, and the only marker used for the detection of prostate cancer. While PSA is highly specific for prostatic tissue, a high PSA serum level is not specific for prostate cancer, since it can be high even in benign prostatic disease. There is a need for a simple exam to define the opportunity of a prostate biopsy in men with an altered PSA levels. Detection of malignant prostatic cells (mCPC) in blood may be a candidate for early detection of prostate cancer (PC). Patients and methods: a group of patients, who underwent an initial prostate biopsy -and also a second one during a follow up - due to suspicion of prostate cancer, were assessed prospectively. The results of the biopsy were compared with malignant circulating prostate cells (mCPC) levels, identified in blood samples drawn prior to the biopsy, using standard immunocytochemistry methods. The objective was to determine the diagnostic ability of mCPC before the first, the second and the third prostate biopsies. Results: In total, 423 consecutive patients participated in the study, with an average age of 65.3 ± 8.9 years and a PSA (median) of 5.28 ng/ml (interquartile range [IQR] 4.36-7.94 ng/ml). Of them, 138 (32.6%) had a prostate cancer detected in the initial biopsy. Tests for the detection of circulating prostatic cells in blood (CPCs) had a sensitivity of 0.89 (95% confidence interval [95% CI: 0.82-0.94) and a specificity of 0.89 (95% CI: 0.84 to 0.92). Of the 423 patients, 125 had a second biopsy, and 57 had a third biopsy. CPCs attained a sensitivity and specificity of 0.89/0.87 and 0.88/0.96 for the second and third biopsy, respectively, with a positive and negative predictive value of 0.65 / 0.97 and 0.88 / 0.95 in the second and third biopsy, in that order. Conclusions: CPCs used in combination with current screening tests -PSA and digital rectal exam- can improve screening effectiveness by reducing the frequency of negative biopsies, as well as the total number of biopsies and their complications. In addition, profitability for the public health system in terms of resource utilization, is positive

Prediction model for early biochemical recurrence after radical prostatectomy based on the Cancer of the Prostate Risk Assessment score and the presence of secondary circulating prostate cells.

OBJECTIVE: To establish a prediction model for early biochemical recurrence based on the Cancer of the Prostate Risk Assessment (CAPRA) score and the presence of secondary circulating prostate cells (CPCs). PATIENTS AND METHODS: We conducted a prospective single-centre study of men who underwent radical prostatectomy as monotherapy for prostate cancer. Clinicopathological findings were used to calculate the CAPRA score. At 90 days after surgery, blood was taken for CPC detection, mononuclear cells were obtained using differential gel centrifugation, and CPCs were identified using immunocytochemistry. A CPC was defined as a cell expressing prostate-specific antigen (PSA) but not CD45. The CPC test results were defined as positive or negative. Patients were followed up for up to 5 years and biochemical recurrence was defined as a PSA level >0.2 ng/mL. The validity of the CAPRA score was calibrated using partial validation, and Cox proportional hazard regression to build three models: a CAPRA score model, a CPC model and a CAPRA/CPC combined model. RESULTS: A total of 321 men, with a mean age of 65.5 years, participated in the study. After 5 years of follow-up the biochemical recurrence-free survival rate was 98.55%. For the model that included CAPRA score there was a hazard ratio (HR) of 7.66, for the CPC model there was an HR of 34.52 and for the combined model there were HRs of 2.60 for CAPRA score and 22.5 for CPC. Using the combined model, 23% of men changed from the low-risk to the high-risk category, or vice versa. CONCLUSION: The incorporation of CPC detection significantly improved the model's discriminative ability in establishing the probability of biochemical recurrence; patients in the high-risk group according to CAPRA score who are negative for CPCs have a much better prognosis. The addition of CPC detection gives clinically significant information to aid the decision on who may be eligible for adjuvant therapy.

El uso de la detección de células prostáticas en la circulación sanguínea podría disminuir el número de biopsias prostáticas necesarias para detectar cáncer prostático / Detection of circulating prostate cells could avoid prostate biopsies

Introducción: El uso del antígeno prostático específico como pesquisa para cáncer prostático ha significado según algunos estudios una disminución en la mortalidad y un cambio hacia las etapas más precoces. Pero, implica también que aproximadamente70 por ciento de los hombres con un APE elevado tendrán una biopsia negativa para cáncer, asumiéndose los riesgos de hemorragia e infección del procedimiento. Presentamos un estudio sobre la detección de células prostáticas en la circulación sanguínea como examen complementario y los resultados de la biopsia prostática. Método y pacientes: A hombres que cumplían con los criterios para someterse a una biopsia prostática, se les tomó una muestra de sangre. Las células mononucleares fueron separadas usando centrifugación diferencial y las células prostáticas detectadas usando anticuerpos monoclonales contra el APE y identificadas con inmunocitoquímica. Los resultados de la presencia o ausencia de las CPCs fueron comparados con los resultados de la biopsia. La biopsia fue dirigida por ecografía y tomada siguiendo la norma estándar en sextante. Resultados: Participaron 358 hombres, de éstos, 91 pacientes cumplieron con los criterios para una biopsia, de los cuales 86 se les tomó una biopsia. La ausencia de CPCs fue asociada con una biopsia negativa en 94,7 por ciento (54/57) y hubo CPCs detectadas en 24/27 (89 por ciento) de los casos con una biopsia positiva para cáncer. En 3 casos biopsia positiva CPC negativa el cáncer fue de bajo grado y localizado. Hubo una sensibilidad de 91,5 por ciento y una especificad de 89,0 por ciento. Conclusiones: Hombres negativo para CPCs tienen una alta posibilidad de una biopsia prostática negativa (94,7 por ciento), en estos hombres es posible postergar la biopsia con un monitoreo cuidadoso del APE sérico. Evitando biopsias no necesarias disminuirán los riesgos asociados al paciente sin aumentar los riesgos de no detectar un cáncer agresivo.

Introduction: The widespread use of PSA screening for prostate cancer has decreased mortality and increased early stage detection. However, approximately 70 percent of biopsies will be negative in men with an increased PSA, incurring in the associated risks of haemorrhage and infection. We report the use of circulating prostate cells (CPCs) as a complementary test and compare the results with the associated prostate biopsy. Patients and Methods: Men fulfilling biopsy criteria had a blood sample taken, the mononuclear cells were separated using differential centrifugation and detected using monoclonal antibodies against PSA and identified using monoclonal antibodies against PSA and identified using immunocytochemistry. Standard ultrasound guided sextant biopsy was used. The presence or absence of CPCs was compared with prostate biopsy results. Results: Of 358 men participating in the study 91 fulfilled biopsy criteria of which 86underwent biopsy. The absence of CPCs was seen in 94.7 percent of cases (54/57) with a negative biopsy. In the 3 remaining cases the CPC negative patient had a low grade local cancer. Overall there was a sensitivity of 91.5 percent and specificity of 89.0 percent. Conclusions: Men with negative CPC have a high probability of a negative biopsy, in these patients the biopsy could be deferred with close monitoring of PSA levels, thus avoiding biopsy complications. This would avoid unnecessary risks without jeopardizing early stage cancer detection.