Currently, there are no reliable prognostic factors to determine which upper tract urothelial carcinoma (UTUC) patients will progress after radical nephroureterectomy (RNU). We aim to evaluate whether liquid-biopsy-based biomarkers (circulating tumor cells (CTCs), cell-free DNA (cfDNA), and circulating tumor DNA (ctDNA)) were able to predict clinical outcomes in localized UTUC patients undergoing RNU. Twenty patients were prospectively enrolled between 2021 and 2023. Two blood samples were collected before RNU and three months later. CTCs and cfDNA were isolated and evaluated using the IsoFlux system and Quant-iT PicoGreen dsDNA kit, respectively. Droplet digital PCR was performed to determine ctDNA status. Cox regression analysis was performed on CTCs, cfDNA, and ctDNA at two different follow-up time points to examine their influence on tumor progression and cancer-specific survival (CSS). During a median follow-up of 18 months, seven (35%) patients progressed and three (15%) died. Multivariate analysis demonstrated that cfDNA levels three months after RNU are a significant predictor of tumor progression (HR = 1.085; p = 0.006) and CSS (HR = 1.168; p = 0.029). No associations were found between CTC enumeration and ctDNA status with any of the clinical outcomes evaluated. The evaluation of cfDNA levels in clinical practice could improve the disease management of UTUC patients.

Objective: To evaluate the performance of Bladder Epicheck® (BE; Nucleix Ltd., Rehovot, Israel) in predicting tumour recurrence and bacillus Calmette–Guérin (BCG) failure during the first year after induction treatment. Patients and methods: Prospective study including 65 patients with non‐muscle‐invasive bladder cancer treated with BCG between 2018 and 2021. Urine samples analysed with BE were collected before and after BCG induction. Logistic binary regression was used to assess the association between clinical and pathological variables and BE results with tumour recurrence and BCG failure during the first year after induction treatment. Results: During follow‐up, 16 (24.6%) patients experienced a bladder cancer event, 11 (68.8%) of which were BCG failure (high‐grade recurrence) and five (31.2%) were low‐grade recurrences. The median (range) time to overall recurrence was 7.3 (3.8–17.4) months. A significant association was found between the risk of tumour recurrence/BCG failure and post‐BCG cystoscopy (odds ratio [OR] 10.0; P < 0.001 and OR 13.1; P < 0.001, respectively), post‐BCG BE result (OR 16.9; P < 0.001 and OR 33.1; P < 0.001, respectively) and pre/post‐BCG EpiScore value variation (OR 14.4; P = 0.001 and OR 7.1; P = 0.018, respectively). A nomogram including these three variables outperformed the Club Urológico Español de Tratamiento Oncológico (CUETO) risk tables to predict any bladder cancer event after BCG induction (area under the curve 95.1% vs 67.1%). Result validation in a larger and independent series is needed. Conclusions: The BE post‐BCG status and variations in EpiScore values can help us identify patients at higher risk of any bladder cancer event and BCG failure promptly. These data can have an impact on disease management.

Prostate cancer recurrence following primary treatment poses a significant clinical challenge, particularly when detected through biochemical recurrence at low PSA levels. Conventional imaging modalities often fail to localize the disease at this early stage. PSMA PET has demonstrated superior sensitivity in detecting recurrent lesions, even in patients with low PSA. Concurrently, liquid biopsy, through analysis of cell-free DNA (cfDNA), offers a minimally invasive approach for monitoring disease. There is scarce evidence about the association between liquid biopsy and PSMA PET/CT findings. This study aimed to assess the correlation between liquid biopsy and tumor burden assessed by PSMA PET/CT in early recurring prostate cancer patients. PSMA PET/CT and liquid biopsies of 32 patients in biochemical recurrence were analyzed. 12 patients (37.5%) had no PSMA PET-measurable disease. Four patients (12.5%) presented local recurrence, seven (21.9%) had recurrence in pelvic lymph nodes, one of whom also had local recurrence. Nine patients (28.1%) presented metastatic recurrence, with or without local or nodal recurrence. PSA levels correlated with molecular imaging data (p < 0.05), including whole body PSMA-TV, whole body PSMA-TL, whole body SUVmean and whole body SUVmax. The mean cfDNA fragment size fraction was inversely correlated with tumour burden measured with whole body PSMA-TV, with a Spearman correlation coefficient of -0.451 and a p-value of 0.009. No correlation was found between cfDNA concentration and PET-PSMA data. This prospective study demonstrated a statistically significant negative correlation between cfDNA fragmentation patterns and PSMA PET/CT volumetric parameters in patients with presumed localized prostate cancer with early biochemical recurrence. These findings underscore the potential of liquid biopsy as a biomarker and a complementary tool to PSMA PET/CT to assess disease progression during the follow-up of these patients.

Cystoscopy remains the gold standard for initial diagnostic assessment and follow-up in bladder cancer (BC) patients. Novel diagnostic tools and biomarkers are needed to minimize reliance on cystoscopies whenever possible. Bladder EpiCheck^® (BE) represents one such approach with encouraging results. However, its diagnostic performance at primary BC diagnosis and its effectiveness across distinct non-muscle-invasive BC (NMIBC) risk groups during follow-up have not yet been described. Prospective case-control study between July 2018 and December 2024 including two cohorts: cohort 1 comprised individuals during primary diagnosis of BC (n = 254); cohort 2 included BC patients in follow-up for disease recurrence (n = 138). Overall, 415 voided urine samples were collected (n = 254 for cohort 1; n = 161 for cohort 2) and analyzed using BE test (Nucleix Ltd). Urine cytology was conducted whenever sufficient residual urine was available. Diagnostic performance of both techniques in detecting BC was compared. Overall, BE sensitivity (SN) for detecting primary BC and recurrent NMIBC was 73% and 55%, respectively. When limited to high-grade (HG) tumours, BE SN increased to 92% and 87%, respectively. BE showed comparable specificity (SP) to cytology in primary (99% vs. 100%) and in recurrent tumours (91% vs. 100%). Diagnostic performance of BE was significantly better than that of cytology in primary (p = 0.013) and recurrent patients (p = 0.023). BE and cytology SN increased progressively across higher NMIBC risk groups. However, BE SN ranged from 47% in low-risk to 93% in very-high risk NMIBC patients while cytology SN ranged from 0% to 79%. Strikingly, EpiScore values were higher in those BC patients in follow-up who progressed into a muscle-invasive disease (p < 0.001), and patients with an EpiScore ≥ 90 were more likely to present high/very-high risk NMIBC or MIBC (OR = 1.93; 95% CI: 1.01-3.67; p = 0.046), ≥T1 stage (OR = 2.00; 95% CI: 1.09-3.71; p = 0.026) and HG tumours (OR = 2.59; 95% CI: 1.27-5.29; p = 0.008). During initial diagnosis of BC, BE showed higher SN with similar SP compared with urine cytology. Diagnostic performance of BE increased gradually across EAU prognostic NMIBC factor risk classification, outperforming cytology in all risk groups. Moreover, EpiScore values are able to identify high risk BC patients, ≥T1 tumours and HG tumours.

Background/Objectives: Currently, the prediction of disease recurrence after radical prostatectomy (RP) in localized prostate cancer (PCa) relies on clinicopathological parameters, which lack accuracy in predicting clinical outcomes. This study focused on evaluating the utility of cfDNA levels and fragmentation patterns as prognostic biomarkers in progressive prostate-specific antigen (PSA) patients, including those with persistent PSA and biochemical recurrence (BR), after primary treatment in localized PCa patients. Methods: Twenty-nine high-risk localized PCa patients were enrolled in the study between February 2022 and May 2023. Blood samples were obtained before robotic RP. cfDNA concentration and fragment size were quantified using the Quant-it PicoGreen dsDNA Assay kit and Agilent 2200 TapeStation System, respectively. Results: The mean PSA value at diagnosis was 9.4 ng/mL. Seven patients (24.1%) had stage pT2 and 22 (75.9%) pT3. Nine patients (31%) had detectable PSA at the first PSA control six weeks after surgery, and four patients (20%) had BR during a mean follow-up of 18.4 months. No associations were found between cfDNA levels or fragmentation patterns and clinicopathological data. Although not statistically significant, patients with detectable PSA levels post-surgery exhibited higher cfDNA levels and shorter fragments compared with those with undetectable PSA. Conclusions: Our study indicated a tendency toward more fragmented cfDNA levels in PCa patients with persistent PSA. Strikingly, biochemical recurrent PCa patients exhibited similar cfDNA levels and fragmentation patterns compared to non-recurrent patients. Further studies exploring liquid biopsy-derived biomarkers in localized PCa patients are needed to elucidate their clinical utility in predicting PSA persistence.

Background: This study aimed to evaluate Bladder EpiCheck (BE) performance in detecting urothelial carcinoma (UC) and HG (high‐grade) events in patients with indeterminate cytology and negative cystoscopy. Methods: This prospective study included 63 patients who presented with indeterminate urinary cytology and negative cystoscopy between October 2022 and April 2024. A repeated urine sample was obtained and analyzed via both urinary cytology and BE test. Patients were followed for 1 year. The diagnostic performance of each test was assessed. Logistic regression models were developed to estimate the risk of UC and HG events on the basis of repeated cytology and BE results. Results: Thirty‐two patients (51%) experienced a UC event, of which 25 (78%) were HG events. Sensitivity (SN) for detecting UC events was significantly higher (80% vs. 17%; p < .001) for BE compared to repeated urinary cytology, whereas repeated cytology demonstrated higher specificity (SP) (59% vs. 96%; p = .02). Similar SN differences were observed for HG disease (80% vs. 25%; p < .001). However, when suspicious for high‐grade urothelial carcinoma was considered a positive cytology result, no statistically significant differences were observed between BE and repeated cytology in either SN or SP. A prognostic model combining repeated cytology and BE outperformed models using baseline cytology combined with either test alone in predicting UC and HG events. Conclusions: In patients with indeterminate urinary cytology and negative cystoscopy, BE provided complementary diagnostic information for the detection of UC events and HG disease. The combination of BE and repeated urinary cytology provided the most accurate risk assessment.