[Effect of different schemes of androgen deprivation therapy on bone mass density in prostate cancer patients]. / Efecto de diferentes esquemas de terapia de deprivación androgénica sobre la densidad mineral ósea de pacientes con cáncer de próstata.

OBJECTIVES: To evaluate the aspects of the basal bone health status in prostate cancer patients. Furthermore, to evaluate in a real-world setting the effect of different schemes (intermittent or continuous) of androgen deprivation therapy (ADT) and the effect of denosumab in bone mass density (BMD). METHODS: Observational, retrospective study of a cohort of prostate cancer patients in treatment with luteinizing hormone-releasing hormone (LH-RH) agonists, evaluated in the rheumatology department of a tertiary center. Demographics, FRAX score, LH-RH treatment scheme, osteoporosis treatment, laboratory data and BMD were collected. Mixed effect regression models to analyze the interaction between LH-RH treatment scheme, denosumab and BMD evolution were used. RESULTS: Eighty-three patients (mean age 71±8years) were included. At the basal evaluation, 16% of patients presented densitometric osteoporosis and 27% of patients presented high fracture risk. Eighty percent of patients had inadequate vitaminD levels. VitaminD >30ng/mL was correlated with higher T-scores. There was no association between LH-RH treatment scheme and BMD evolution, however there was a positive association with denosumab. CONCLUSION: A high proportion of patients presented elevated fracture risk or inadequate vitaminD levels, not previously recognized. Bone health assessment and fracture risk evaluation are convenient in these patients. In a real-world setting, the effect of denosumab in BMD is detected, however the effect of intermittent LH-RH schema treatment is less evident.

LC-MS metabolomics of urine reveals distinct profiles for non-muscle-invasive and muscle-invasive bladder cancer.

PURPOSE: Bladder cancer (BC) is among the most frequent malignancies worldwide. Novel non-invasive markers are needed to diagnose and stage BC with more accuracy than invasive procedures like cystoscopy. To date, no study has identified urine metabolites characteristic of all BC stages. To discover novel urine metabolomic profiles to diagnose and stage non-muscle-invasive (NMIBC) and muscle-invasive (MIBC) patients using mass spectrometry-based metabolomics. METHODS: We prospectively recruited 198 BC patients and 98 age- and sex-matched healthy volunteers without evidence of renal or bladder condition confirmed by ultrasound, from whom we collected a first morning urine sample (before surgery in patients). In a discovery stage, an untargeted metabolomic analysis was conducted in urine samples of a selection of 64 BC patients (19 TaG1, 11 TaG3, 20 T1G3, 12 T2G3, 1 T2G2, 1 T3G3) and 20 controls to identify dysregulated metabolites. Next, after exhaustive multivariate analysis, confirmed dysregulated metabolites were validated in an independent cohort of 134 BC patients (19 TaG1, 62 TaG2, 9 TaG3, 15 T1G2, 16 T1G3, 4 T2G2, 9 T2G3) and 78 controls. RESULTS: We validated p-cresol glucuronide as potential diagnostic biomarker for BC patients compared to controls (AUC = 0.79). For NMIBC, p-cresol glucuronide was valuable as staging biomarker (AUC = 0.803). And among NMIBCs, p-coumaric acid may be a potential specific staging biomarker for the TaG1 NMIBC; however, future validation experiments should be conducted once the precise version of the standard is commercially available. Remarkably, for MIBC we validated spermine as potential specific staging biomarker (AUC = 0.882). CONCLUSION: Ours is the first metabolomics study conducted in urine of a thoroughly characterized cohort comprising all stages of NMIBC, MIBC and healthy controls in which we identified non-invasive diagnostic and staging biomarkers. These may improve BC management, thus reducing the use of current harmful diagnostic techniques.

Nuclear-medicine probes: Where we are and where we are going.

Nuclear medicine probes turned into the key for the identification and precise location of sentinel lymph nodes and other occult lesions (i.e., tumors) by using the systemic administration of radiotracers. Intraoperative nuclear probes are key in the surgical management of some malignancies as well as in the determination of positive surgical margins, thus reducing the extent and potential surgery morbidity. Depending on their application, nuclear probes are classified into two main categories, namely, counting and imaging. Although counting probes present a simple design, are handheld (to be moved rapidly), and provide only acoustic signals when detecting radiation, imaging probes, also known as cameras, are more hardware-complex and also able to provide images but at the cost of an increased intervention time as displacing the camera has to be done slowly. This review article begins with an introductory section to highlight the relevance of nuclear-based probes and their components as well as the main differences between ionization- (semiconductor) and scintillation-based probes. Then, the most significant performance parameters of the probe are reviewed (i.e., sensitivity, contrast, count rate capabilities, shielding, energy, and spatial resolution), as well as the different types of probes based on the target radiation nature, namely: gamma (γ), beta (ß) (positron and electron), and Cherenkov. Various available intraoperative nuclear probes are finally compared in terms of performance to discuss the state-of-the-art of nuclear medicine probes. The manuscript concludes by discussing the ideal probe design and the aspects to be considered when selecting nuclear-medicine probes.

Identification of miR-20a-5p as Robust Normalizer for Urine microRNA Studies in Renal Cell Carcinoma and a Profile of Dysregulated microRNAs.

Renal cell carcinoma (RCC) is the third most frequent urinary malignancy and one of the most lethal. Current diagnostic and follow-up techniques are harmful and unspecific in low-grade tumors. Novel minimally invasive markers such as urine microRNAs (miRNAs) are under study. However, discrepancies arise among studies in part due to lack of consent regarding normalization. We aimed to identify the best miRNA normalizer for RCC studies performed in urine samples together with a miRNA profile with diagnostic value and another for follow-up. We evaluated the performance of 120 candidate miRNAs in the urine of 16 RCC patients and 16 healthy controls by RT-qPCR followed by a stability analysis with RefFinder. In this screening stage, miR-20a-5p arose as the most stably expressed miRNA in RCC and controls, with a good expression level. Its stability was validated in an independent cohort of 51 RCC patients and 32 controls. Using miR-20a-5p as normalizer, we adjusted and validated a diagnostic model for RCC with three miRNAs (miR-200a-3p, miR-34a-5p and miR-365a-3p) (AUC = 0.65; Confidence Interval 95% [0.51, 0.79], p = 0.043). let-7d-5p and miR-205-5p were also upregulated in patients compared to controls. Comparing RCC samples before surgery and fourteen weeks after, we identified let-7d-5p, miR-152-3p, miR-30c-5p, miR-362-3p and miR-30e-3p as potential follow-up profile for RCC. We identified validated targets of most miRNAs in the renal cell carcinoma pathway. This is the first study that identifies a robust normalizer for urine RCC miRNA studies, miR-20a-5p, which may allow the comparison of future studies among laboratories. Once confirmed in a larger independent cohort, the miRNAs profiles identified may improve the non-invasive diagnosis and follow-up of RCC.

Human adipose-derived mesenchymal stem cells accelerate decellularized neobladder regeneration.

Decellularized natural bladder matrices (neobladders) represent an exciting means to regenerate the bladder following bladder cancer-associated cystectomy. In this study, we compare the evolution of decellularized matrices with recellularized matrices by seeding it with human adipose-derived mesenchymal stem cells (ADSC) after implantation following partial cystectomy in rats. We discovered significant anatomical differences since 10 days after neobladder implantation with the ADSC-containing matrices promoting a significant recovery of mature p63- and cytokeratin 7-positive urothelium. We also discovered significantly induced expression of the vimentin mesoderm marker in the submucosal layer in ADSC-seeded matrices. Interestingly, we found a higher expression of smooth muscle actin in transversal and longitudinal smooth muscle layers with ADSC-seeded matrices. Furthermore, ADSC also showed increased vascularization and nerve innervation of the neobladder as determined by the distribution of CD31 and S100ß reactivity, respectively. We believe that ADSC and their paracrine-acting pro-regenerative secretome within decellularized matrices represent an efficient bladder substitution strategy; however, we require a fuller understanding of the mechanisms involved before clinical studies can begin.

[Sentinel lymph node selective biopsy in prostate cancer.] / Ganglio centinela en cáncer de próstata.

OBJECTIVE: To validate the sentinel lymph node selective biopsy (SLNSB) in the staging of Prostate Cancer with Briganti Index > 5 by comparison with extended lymphadenectomy (ePLND) in a prospective longitudinal study. METHODS: SLNSB has been performed in 84 patients, the first 70 by injection of nanocoloids marked with Tc99m and preoperative SPECT-CT, and in the last 14 with mixed radiotracer (99mTc + ICG). After laparoscop ic removal of sentinel nodes all patients underwent an ePLND. RESULTS: SPECT-CT showed radiotracer deposits outside the territory of the ePLND in 76% of patients and laparoscopic gamma probe in 57%. The median number of sentinel nodes removed was 5.2 with a total average number of lymph nodes removed of 22. In all cases with metastatic nodes (28% in the series) there was at least one positive sentinel node but metastatic sentinel nodes outside of the territory of the ePLND were found in 6/24 patients (25%). The sensitivity, specificity, PPV and NPV of 99mTc were 100%, 96.07%, 90.47% and 100%, respectively. In 5 out of 14 patients with mixed radiotracer, lymph node involvement was detected. In all of them there was at least one sentinel node affected with 99mTc, and only 3 showed fluorescence with 100% sensitivity and 100% NPV for 99mTc and 60% sensitivity and 77.77% NPV for ICG. CONCLUSION: The SLNSB with 99mTc has a high sensitivity and a VPN of 100%, increasing the identification of lymphatic metastases outside the territory of the ePLND. Fluorescence can facilitate the visualization of the sentinel nodes when they have been previously located by the SPECT-CT, although the sensitivity and the NPV of the ICG are lower than that of the 99mTc.

OBJETIVO: Validar la biopsia selectiva de ganglio centinela (BSGC) en la estadificación del Cáncer de Próstata con Indice de Briganti > 5 mediante comparación con la linfadenectomía extendida (LFDe) en un estudio prospectivo longitudinal.MÉTODOS: Se ha realizado BSGC a 84 pacientes, los 70 primeros mediante inyección de nanocoloides marcados con Tc99m y SPECT-TC preoperatoria, y en los 14 últimos con radiotrazador mixto (Tc99m + ICG). A todos los pacientes tras la extracción laparoscópica de los ganglios centinelas se les realizó una LFDe. RESULTADOS: La SPECT-TC mostró depósitos del radiotrazador fuera del territorio de la LFDe en el 76% de los pacientes y la gammasonda laparoscópica en el 57%.La media de ganglios centinelas extraídos fue 5,2 con una media total de ganglios linfáticos extraídos de 22. En todos los casos con ganglios metastáticos (28% de la serie) hubo, al menos, un ganglio centinela positivo, encontrando ganglios centinela metastásicos fuera del territorio de la LFDe en 6/24 pacientes (25%). La sensibilidad, especificidad, VPP y VPN del Tc99m fue del 100%, 96,07%, 90,47% y 100%, respectivamente. En 5 de los 14 pacientes con radiotrazador mixto se detectó afectación ganglionar. En todos ellos hubo como mínimo un ganglio centinela afecto con Tc99m y sólo 3 mostraron fluorescencia, con sensibilidad del 100% y VPN del 100% para el Tc99m y sensibilidad del 60% y VPN del 77,77% para el ICG.CONCLUSIÓN: La BSGC con Tc99m tiene una alta sensibilidad y un VPN del 100%, aumentando la identificación de metástasis linfáticas fuera del territorio de la LFDe. La fluorescencia puede facilitar la visualización de los centinelas cuando se tiene una localización previa de los mismos con el SPECT-TAC, aunque la sensibilidad y el VPN del ICG es inferior al del Tc99m.

Protein Corona over Mesoporous Silica Nanoparticles: Influence of the Pore Diameter on Competitive Adsorption and Application to Prostate Cancer Diagnostics.

Diagnostic tests based on proteomics analysis can have significant advantages over more traditional biochemical tests. However, low molecular weight (MW) protein biomarkers are difficult to identify by standard mass spectrometric analysis, as they are usually present at low concentrations and are masked by more abundant resident proteins. We have previously shown that mesoporous silica nanoparticles are able to capture a predominantly low MW protein fraction from the serum, as compared to the protein corona (PC) adsorbed onto dense silica nanoparticles. In this study, we begin by further investigating this effect using liquid chromatography-mass spectrometry (LC-MS)/MS and thermogravimetric analysis (TGA) to compare the MW of the proteins in the coronas of mesoporous silica nanoparticles with the same particle size but different pore diameters. Next, we examine the process by which two proteins, one small and one large, adsorb onto these mesoporous silica nanoparticles to establish a theory of why the corona becomes enriched in low MW proteins. Finally, we use this information to develop a novel system for the diagnosis of prostate cancer. An elastic net statistical model was applied to LC-MS/MS protein coronas from the serum of 22 cancer patients, identifying proteins specific to each patient group. These studies help to explain why low MW proteins predominate in the coronas of mesoporous silica nanoparticles, and they illustrate the ability of this information to supplement more traditional diagnostic tests.

Bioengineering Approaches for Bladder Regeneration.

Current clinical strategies for bladder reconstruction or substitution are associated to serious problems. Therefore, new alternative approaches are becoming more and more necessary. The purpose of this work is to review the state of the art of the current bioengineering advances and obstacles reported in bladder regeneration. Tissue bladder engineering requires an ideal engineered bladder scaffold composed of a biocompatible material suitable to sustain the mechanical forces necessary for bladder filling and emptying. In addition, an engineered bladder needs to reconstruct a compliant muscular wall and a highly specialized urothelium, well-orchestrated under control of autonomic and sensory innervations. Bioreactors play a very important role allowing cell growth and specialization into a tissue-engineered vascular construct within a physiological environment. Bioprinting technology is rapidly progressing, achieving the generation of custom-made structural supports using an increasing number of different polymers as ink with a high capacity of reproducibility. Although many promising results have been achieved, few of them have been tested with clinical success. This lack of satisfactory applications is a good reason to discourage researchers in this field and explains, somehow, the limited high-impact scientific production in this area during the last decade, emphasizing that still much more progress is required before bioengineered bladders become a commonplace in the clinical setting.

Therapeutic potential of human adipose-derived stem cells (ADSCs) from cancer patients: a pilot study.

Mesenchymal stem cells from adipose tissue (ADSCs) are an important source of cells for regenerative medicine. The therapeutic effect of culture-expanded adipose derived stem cells has been shown; however, optimal xeno-free culture conditions remain to be determined. Cancer patients, specifically those undergoing invasive surgery, constitute a subgroup of patients who could benefit from autologous stem cell transplantation. Although regenerative potential of their ADSCs could be affected by the disease and/or treatment, we are not aware of any study that has evaluated the therapeutic potential of ADSCs isolated from cancer patients in reference to that of ADSCs derived from healthy subjects. Here we report that ADSCs isolated from subabdominal adipose tissue of patients with urological neoplasms yielded similar growth kinetics, presented equivalent mesenchymal surface markers and showed similar differentiation potential into distinct mesodermal cell lineages: adipocytes, chondroblasts and osteoblasts than ADSCs isolated from adipose tissue of age-matched non-oncogenic participants, all under xeno-free growth culture conditions. Molecular karyotyping of patient expanded ADSCs genomes showed no disease-related alterations indicating their safety. In addition, vesicles <100 nm identified as exosomes (EXOs) which may be at least partly responsible for the attributed therapeutic paracrine effects of the ADSCs were effectively isolated from ADSCs and showed equivalent miRNA content regardless they were derived from cancer patients or non-oncogenic participants indicating that the repair capabilities of xeno-free expanded ADSCs are not compromised by patient condition and therefore their xeno-free culture expanded ADSCs should be suitable for autologous stem cell transplantation in a clinical setting.

Prognostic value of p53, Ki-67, microstaging and microvessel density in pT1G3 bladder tumors: creation of risk groups for progression.

OBJECTIVE: New predictive factors for bladder tumor progression have been analyzed in many publications, often with contradictory results. Very few papers have referred specifically to T1G3 tumors. Our objective was to find new, clinically useful markers which either alone or in association with classical prognostic factors would allow the early selection of the correct therapeutic approach. MATERIAL AND METHODS: This was a retrospective study of 83 patients with T1G3 bladder tumors who were initially treated with transurethral resection + bacillus Calmette-Guérin therapy, with a minimum follow-up period of 3 years. We analyzed eight variables. New factors considered were: the level of submucosal invasion; microvessel density; and immunostaining for Ki-67 and p53. Independent prognostic variables for progression were established using logistic regression analysis, and risk groups were created from mathematical models. RESULTS: Five variables were determined as unfavorable: tumor multiplicity; tumor size >3 cm; carcinoma in situ; T1b substage; and p53 positivity. The first three factors predicted progression in only 32% of cases, while the addition of the new prognostic factors (T1b substage and p53 positivity) increased this rate to 65%. We established four risk groups, with rates of progression of 67% and 100% in the high-risk and very high-risk groups, respectively. For inclusion in these groups, both new predictive factors had to be unfavorable; if either one were absent then the three classical factors had to be present. CONCLUSIONS: Microstaging and p53 positivity have a prognostic value for predicting progression in T1G3 tumors, providing 33% more information than that obtained with classical prognostic factors alone. The application of mathematical models identifies risk groups and allows the use of an early and more aggressive treatment.