The sentinel node with technetium-99m for prostate cancer. A safe and mature new gold standard?

BACKGROUND: The objective was to carry out a prospective study to compare the current extended pelvic lymph node dissection (ePLND) to the sentinel node (SN) technique with 99mTcnanocolloid. METHODS: We conducted a prospective study between January 2013 and May 2020. In the first 74 patients, 99mTc-nanocolloid was used. Then from June 2017 onwards, in 38 patients we used a combined radiotracer prepared by adding indocyanine green (ICG). A preoperative SPECT/CT was also performed to check on the SNs. We extracted the SNs guided by a laparoscopic gamma-ray detection probe and/or a fluorescence camera. RESULTS: We included 112 patients with a Briganti nomogram-assessed risk of 5% or more. In 4 out of the total, the radiotracer did not migrate. The mean number of extracted nodes was 21.56 (13.46-29.71) and the mean of extracted SNs was 5.17 (1.83-8.51) (P<0.001). The technique that registered the most nodes with high activity was SPECT/CT, with an average of 4.33 nodes (2.42-6.23) (P<0.001). We found SNs outside the template in 78% of the patients. A total of 46% of the complications were related to ePLND. The SN biopsy showed a sensitivity of 100%, specificity of 97.5%, PVV of 92.86%, and NPV of 100%. CONCLUSIONS: Our results prove that ePLND is a technique with significant morbidity; up to 46% of the complications were related to the ePLND. The SN surgery showed great accuracy in detecting metastases due to the SPECT/CT and a lower rate of complications than ePLND.

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.

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.

Urinary microRNAs: Looking for a New Tool in Diagnosis, Prognosis, and Monitoring of Renal Cancer.

PURPOSE OF REVIEW: Renal cell carcinoma (RCC) is the third most common urologic malignancy. First symptoms are usually unspecific and belated causing the stages to be high when diagnosed. As compensation, incidental detection of RCC by abdominal imaging techniques for other medical purposes is a reality that favours a decrease in the stage of new diagnosed tumours. Therefore, identifying novel predictive biomarkers for diagnosis, progression and prognosis of RCC is fundamental. RECENT FINDINGS: To date, several studies have demonstrated that microRNAs (miRNAs) play a role in the particular scenario of urologic tumors, and alterations at miRNA level are involved in the initiation, progression and metastases formation of renal cancer. In the present review, we have summarized the up­to­date preliminary clinical works on the role of urinary miRNA profiling in RCC, including an evaluation of its value as a potential biomarker for diagnosis, prognosis and follow up of RCC patients.

Surgical Treatment of Completely Endophytic Renal Tumor: a Systematic Review.

PURPOSE OF REVIEW: An endophytic renal tumor represents a special surgical challenge in terms of location and safe removal. For this reason we wanted to review the existing literature on this subject. RECENT FINDINGS: In high-activity robotic centers, robot-assisted partial nephrectomy (RAPN) is a safe and efficacious surgical approach for completely endophytic renal tumors. As research innovation, the application of the radio-guided occult lesion localization technique (ROLL) facilitates the location and complete excision of the tumor during surgery. There are few studies that specifically report the experience with completely endophytic renal tumors. The endophytic tumor is usually smaller than exophytic. Frequently it represents a high complexity value in the different Score systems reported in the last decade. This surgery should be performed by experienced urologists regardless of the surgical approach they prefer (open, laparoscopic, or robotic). It is necessary to develop new techniques for intraoperative easy localization and intraoperative evaluation of surgical margins.

Central Body Fat Mass Measured by Bioelectrical Impedanciometry But Not Body Mass Index Is a High-Grade Prostate Cancer Risk Factor.

OBJECTIVE: To evaluate the association between body fat mass distribution measured by bioelectrical impedanciometry (BEI) and high-grade prostate cancer (HGPC). METHODS: We prospectively analyze 323 patients who underwent prostate biopsy. BEI was performed prior to biopsy. Prostate cancer (PC) was stratified according to D'Amico classification. For univariate analysis, Student t test was done. For multivariate analysis, bivariate logistic regression was performed using PSA, body mass index (BMI), percentage central body fat, percentage total body fat, and visceral fat as explicative variables for the diagnosis of HGPC. RESULTS: PC was found in 134 patients. Thirty seven (27.2%) were HGPC. This group had higher age, PSA, and percentage central body fat (p = 0.001, p = 0.001, p = 0.04). BMI showed no association with HRPC. Age, PSA, and percentage central body fat (OR 1,123, 95% CI 1,022-1,233, p = 0.001) were independent risk factors. CONCLUSIONS: Central body fat measured by BEI could explain the association between obesity and HGPC better than BMI suggesting the use of this technique to study body fat distribution.

Bladder cancer patients have increased NETosis and impaired DNaseI-mediated NET degradation that can be therapeutically restored in vitro.

Background: Neutrophils, key players of the immune system, also promote tumor development through the formation of neutrophil extracellular traps (NETs) in a process called NETosis. NETs are extracellular networks of DNA, histones and cytoplasmic and granular proteins (calprotectin, myeloperoxidase, elastase, etc.) released by neutrophils upon activation. NETs regulate tumor growth while promoting angiogenesis and invasiveness, and tumor cells also stimulate NETosis. Although NETosis seems to be increased in cancer patients, an increase of NETs in plasma may also be mediated by an impaired degradation by plasma DNaseI, as evidenced in several immunological disorders like lupus nephritis. However, this has never been evidenced in bladder cancer (BC) patients. Herein, we aimed to evaluate the occurrence of increased NETosis in plasma and tumor tissue of BC patients, to ascertain whether it is mediated by a reduced DNaseI activity and degradation, and to in vitro explore novel therapeutic interventions. Methods: We recruited 71 BC patients from whom we obtained a plasma sample before surgery and a formalin-fixed paraffin embedded tumor tissue sample, and 64 age- and sex-matched healthy controls from whom we obtained a plasma sample. We measured NETs markers (cell-free fDNA, calprotectin, nucleosomes and neutrophil elastase) and the DNaseI activity in plasma with specific assays. We also measured NETs markers in BC tissue by immunofluorescence. Finally, we evaluated the ability of BC and control plasma to degrade in vitro-generated NETs, and evaluated the performance of the approved recombinant human DNaseI (rhDNaseI, Dornase alfa, Pulmozyme®, Roche) to restore the NET-degradation ability of plasma. In vitro experiments were performed in triplicate. Statistical analysis was conducted with Graphpad (v.8.0.1). Results: NETosis occurs in BC tissue, more profusely in the muscle-invasive subtype (P<0.01), that with the worst prognosis. Compared to controls, BC patients had increased NETosis and a reduced DNaseI activity in plasma (P<0.0001), which leads to an impairment to degrade NETs (P<0.0001). Remarkably, this can be therapeutically restored with rhDNaseI to the level of healthy controls. Conclusion: To the best of our knowledge, this is the first report demonstrating that BC patients have an increased NETosis systemically and in the tumor microenvironment, in part caused by an impaired DNaseI-mediated NET degradation. Remarkably, this defect can be therapeutically restored in vitro with the approved Dornase alfa, thus Pulmozyme® could become a potential therapeutic tool to locally reduce BC progression.

Circulating Cell-Free DNA in Liquid Biopsies as Potential Biomarker for Bladder Cancer: A Systematic Review.

Bladder cancer (BC) is among the most frequent cancer types in the world and is the most lethal urological malignancy. Presently, diagnostic and follow-up methods for BC are expensive and invasive. Thus, the identification of novel predictive biomarkers for diagnosis, progression, and prognosis of BC is of paramount importance. To date, several studies have evidenced that cell-free DNA (cfDNA) found in liquid biopsies such as blood and urine may play a role in the particular scenario of urologic tumors, and its analysis may improve BC diagnosis report about cancer progression or even evaluate the effectiveness of a specific treatment or anticipate whether a treatment would be useful for a specific patient depending on the tumor characteristics. In the present review, we have summarized the up-to-date studies evaluating the value of cfDNA as potential diagnostic, prognostic, or monitoring biomarker for BC in several biofluids.

Identification of miR-29c-3p as a Robust Normalizer for Urine microRNA Studies in Bladder Cancer.

Bladder cancer (BC) is among the most frequent malignancies worldwide, being the most expensive cancer to treat and monitor and the most lethal urological cancer. Urine microRNAs (miRNAs) have been proposed as novel non-invasive biomarkers to early diagnose and monitor BC patients in order to avoid the performance of current aggressive diagnostic techniques. However, huge discrepancies arise among studies mainly due to the lack of standardization in the normalization, a crucial step in all miRNA studies. Our aim was to identify the best miRNA normalizer for miRNA studies in urine of BC patients. We evaluated the performance of 110 candidate miRNAs in urine of 35 BC patients and 15 healthy controls by Real Time quantitative Polymerase Chain Reaction (RT-qPCR) followed by a stability analysis with RefFinder. In this screening stage, miR-29c-3p arose as the most stably expressed miRNA in BC and controls, with a good expression level. Stability of miR-29c-3p expression was validated in an independent cohort of 153 BC patients and 57 controls. Finally, we evaluated the robustness of miR-29c-3p as normalizer in the expression study of miR-200c-3p, a potential diagnostic marker for BC. We propose miR-29c-3p as a normalizer for miRNA studies in BC urine. This is the first study that characterizes a reliable normalizer that may allow the comparison of future urine miRNA studies as non-invasive biomarkers for BC diagnosis and monitoring.

Urine metabolomic analysis in clear cell and papillary renal cell carcinoma: A pilot study.

Renal cell carcinoma (RCC) is one of the most lethal type of tumors and is twice more frequent in men than in women. Initial symptoms are unspecific and belated thus increasing mortality. Moreover, current diagnostic and monitoring tools are harmful for the patient and unspecific in low-grade tumors. Therefore, novel minimally-invasive markers are needed to diagnose and monitor RCC patients. Urine represents the ideal sample source of non-invasive biomarkers for RCC. In our study we aimed to identify a urine metabolomic profile characteristic of RCC patients with diagnostic purposes and also to identify a profile with prognostic value. By an UPLC-Q-ToF MS untargeted metabolomic analysis, we compared the metabolomic profile of 23 RCC patients (14 clear cell RCC and 9 papillary RCC) before surgery and that of 23 healthy controls. Additionally, for the first time, we compared the metabolomic profile of these RCC patients pre-nephrectomy and 3 months and one year post-nephrectomy. We identified the dysregulated metabolomic variables by querying their exact mass against those presented in the Metlin and Human Metabolome Database. Next, we experimentally confirmed their identity. Both RCC subtypes showed similar metabolomic patterns at all stages. 51 metabolomic variables were significantly increased in RCC compared to controls and, among them, 4 were selected as potential discriminant metabolites between groups. We could experimentally confirm the identity of p-cresol glucuronide thus describing for the first time an increase in this metabolite in urine of RCC patients (fold change = 2.922, P = .012). Additionally, we confirmed that no metabolomic differences occur 3 months post-nephrectomy in RRC, while 188 variables were significantly increased one year post-nephrectomy. Of the 15 most discriminant metabolomic variables, we could experimentally confirm the identity of isobutyryl-l-carnitine (fold change = 2.098, P = .004) and l-proline betaine (fold change = 3.328, P = .004), for the first time. In summary, we have identified urine p-cresol glucuronide as potential diagnostic marker for RCC and isobutyryl-l-carnitine and l-proline betaine as potential prognostic markers. When confirmed in an independent cohort of RCC patients, these markers may improve the diagnosis and monitoring of RCC patients thus reducing current harmful diagnostic procedures. SIGNIFICANCE: The high-radiation dose of current imaging techniques available to diagnose and monitor renal cell carcinoma (RCC) are harmful for the patient and unspecific in low-grade tumors. Our untargeted metabolomic analysis carried out in urine samples from RCC patients and healthy individual reveals p-cresol glucuronide as potential diagnostic marker for RCC. Additionally, the analysis of RCC urine samples one year post-nephrectomy reveals isobutyryl-l-carnitine and l-proline betaine as potential prognostic markers. These novel non-invasive urine biomarkers may improve RCC management thus reducing the use of current harmful diagnostic techniques.

[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.

Application of the Radio-Guided Occult Lesion Localization Technique for Renal Lumpectomy: From the Laboratory to the Patient.

Incidence of small renal masses is increasing, and preservation of healthy kidney tissue along surgical removal is a challenge, especially if the tumor is small or nonexophytic. To facilitate the location and complete excision of the tumor during surgery, we have implemented the application of the radio-guided occult lesion localization technique from initial idea to clinical application.