Carmen Jerónimo

Genitourinary tumors comprise distinct entities with different epidemiological background, molecular alterations, and clinical outcomes, underlining the need for precision diagnostics. Recent advances in molecular technologies have facilitated the increasing use of molecular testing in routine diagnostics. These tests have important roles in diagnosis, risk stratification, and therapeutic tailoring, and are both tissue-based and liquid biopsy-based. Examples include 1) high-end (epi)genomic arrays, such as mRNA-based arrays for prostate cancer; 2) noninvasive circulating biomarkers (DNA mutations or DNA methylation) in bladder cancer, and serum/plasma microRNA (micrcoRNA-371a-3p) testing in testicular germ cell tumors; 3) as well as targeted assays focused on immunohistochemistry and fluorescence in situ hybridization (FISH) for renal and penile cancers. In this review, we cover the application of these tests in routine practice at a molecular pathology lab. These tests are at different stages of development and establishment in routine practice. As molecular testing increases, so does the relevance of pre-analytics, sample selection, and morphological assessment, which are key for achieving reliable results. Integration of molecular tests is set to contribute to precision medicine in genitourinary cancers, contributing to better selection of patients for targeted therapies. Centralization of techniques, human resources, and resources is critical to establish effective molecular testing frameworks for the community.

Unusual trophoblastic tumors of germ cell origin-epithelioid trophoblastic tumor (ETT), unclassified trophoblastic tumor (UTT), cystic trophoblastic tumor (CTT), and monophasic choriocarcinoma (MChorio)-are rare and mostly encountered postchemotherapy. Their relationship to other germ cell tumor types and gestational tumors remains uncertain. We used microRNA‑371~373 levels and DNA methylation profiling to compare nonchoriocarcinoma germ cell-derived trophoblastic tumors to gestational trophoblastic neoplasms and postpubertal-type teratoma. A total of 36 trophoblastic tumors of germ cell origin were included (biphasic choriocarcinoma [n=16], MChorio [n=4], CTT [n=9], UTT [n=6], and ETT [n=1]). MicroRNA‑371~373 levels were assessed by RT‑qPCR on germ cell-derived trophoblastic tumors and compared with nontrophoblastic testicular germ cell tumors and gestational trophoblastic tumors (n=50). DNA methylation profiling was performed using EPIC array on germ cell-derived trophoblastic tumors and compared with postpubertal-type teratomas and gestational trophoblastic tumors (n=28). Nonchoriocarcinoma trophoblastic tumors of germ cell origin showed significantly lower miR‑371a‑3p levels than germ cell-derived biphasic choriocarcinomas ( P =0.0002). Compared with teratoma, differentiating trophoblastic tumors retained higher miR‑371a‑3p levels ( P =0.0129). Analyses performed using the top differentially methylated probes revealed that CTT, UTT, and ETT of germ cell origin clustered with teratoma, while choriocarcinomas of germ cell origin clustered with gestational trophoblastic tumors. DNA methylation patterns showed increasing differentiation from choriocarcinoma toward teratoma-like profiles. Our results suggest that germ-cell-derived ETT, UTT, and CTT represent “differentiated” trophoblastic tumor phenotypes with similarities to teratoma, whereas germ-cell-derived choriocarcinomas resemble gestational trophoblastic tumors.

Chromatin accessibility has been shown to stratify prostate cancer (PCa) into clinically relevant subtypes. Chromatin-modifying enzymes, such as histone methyltransferases, modulate nucleosome compactness, and specifically, G9a was found to be overexpressed in PCa. G9a inhibition with CM‑272 demonstrated potent anti-tumor effects, including in PCa in vitro models, although its transcriptomic consequences remain largely unexplored. Here, we used the BM18 PCa patient‑derived xenograft (PDX) model to validate the therapeutic efficacy of CM‑272 and to understand the underlying molecular mechanisms. In vivo G9a inhibition reprogrammed androgen receptor (AR) chromatin distribution in an exposure-time-dependent manner. Short-term treatment displaced AR from key oncogenic loci, associating with reduced expression. Conversely, prolonged exposure decreased H3K9me2 while enhancing AR binding to the promoter of tumor suppressor genes, thereby associating with increased expression. Integration with a published PCa chromatin dataset revealed increased AR scores, suggesting a potential androgen-responsive state. This reprogramming enabled CM‑272 to synergize with enzalutamide in vitro . Moreover, TRAIL pathway enrichment and cell cycle arrest were observed upon short-term treatment, thus sensitizing BM18 organoids to TRAIL‑mediated apoptosis. Collectively, these findings identify G9a as a therapeutically actionable regulator in high-risk, AR-dependent PCa and revealed context-dependent vulnerabilities that may be exploited to delay progression to castration-resistant disease.

High‐risk non‐muscle invasive bladder cancer (NMIBC) is a common urological malignancy with a high propensity for recurrence (30–40% of patients) despite treatment with Bacillus Calmette–Guérin (BCG) instillations. Identifying patients who are likely to respond or not to BCG therapy before treatment initiation may significantly improve patient outcomes and minimise unnecessary treatments. This systematic review critically assessed the existing literature on the potential of non‐invasive biomarkers to predict BCG failure in NMIBC patients. A comprehensive bibliographic search on PubMed, Scopus and Medline databases was conducted to identify relevant studies published up to July 7, 2025. After a thorough screening and selection process, 72 studies were included in the final analysis, 58 of which focused on prognostic markers, 6 on predictive markers, and 8 examined both. Moreover, 55 studies focused on tissue, while 28 studied liquid biopsy‐derived biomarkers, showcasing their ability to reflect the dynamic characteristics of the tumour and its microenvironment. The most frequently studied biomarkers comprised p53, Ki‐67 and pRB. Furthermore, several immunological markers were also explored, highlighting the current challenge of BCG therapy prediction biomarkers. Despite its valuable insights, this systematic review faced limitations within the included studies, such as issues related to reproducibility, feasibility, and small sample sizes. Nonetheless, the findings indicate that a wide array of molecules show potential as prognostic and/or predictive markers for BCG failure, which may improve the clinical management of this complex disease.

Testicular cancer is the most common cancer in young adult men, and its incidence is increasing globally, with testicular germ cell tumors (TGCT) being the most frequent subtype. These tumors show remarkable sensitivity to cisplatin, but there is a small subset of patients that develop resistance to the therapy, experiencing low quality of life and having no treatment options. Nowadays, there are a number of models that are useful for studying TGCT tumorigenesis and identifying novel targeted therapies for these patients. Thus, in this review, we aim to provide a comprehensive overview of the in vitro and in vivo TGCT models that are currently available, and also to summarize the main findings that have been made in the last years regarding preclinical and clinical studies of novel targeted therapies in TGCT patients.

Breast cancer is classified into distinct molecular subtypes, including Luminal A, Luminal B, HER2-enriched, Basal-like, and Claudin-low. While traditional studies mostly use 2D cell cultures, 3D models better mimic in vivo tumor conditions. In this study, we generated and characterized 3D spheroids from breast cancer cell lines representing different molecular subtypes. Morphologically, spheroids were either compact (MCF-7/AZ, T47D, BT474, MDA-IBC-3, BT-20, SUM149PT) or loosely adhered (MDA-MB-468, SK-BR-3, MDA-MB-231), while retaining key parental subtype biomarkers. Cell viability decreased with increasing spheroid size, but apoptotic cCasp3 staining was restricted to Basal-like spheroids. Compact spheroids expressed E- and/or P-cadherin, indicating epithelial or epithelial–mesenchymal transition (EMT) hybrid traits, while loose spheroids showed vimentin expression linked to a mesenchymal phenotype. In conclusion, EMT-associated features, rather than intrinsic molecular subtype, may contribute to 3D spheroid architecture of breast cancer cell lines.

There is an urgent need for the development of innovative diagnostic and therapeutic strategies in bladder cancer (BlCa) management, due to its significant

Breast cancer is classified into distinct molecular subtypes, including Luminal A, Lu-minal B, HER2-enriched, Basal-like, and Claudin-low. While traditional studies mostly use 2D cell cultures, 3D models better mimic in vivo tumour conditions. In this study, we generated and characterized 3D multicellular tumour spheroids (MCTS) from breast cancer cell lines representing different molecular subtypes. Morphologically, spheroids were either compact (MCF-7/AZ, T47D, BT474, MDA-IBC-3, BT-20, SUM149PT) or loosely adhered (MDA-MB-468, SK-BR-3, MDA-MB-231), while retain-ing key parental subtype biomarkers. Cell viability decreased with increasing spheroid size, but apoptotic cCasp3 staining was restricted to basal-like spheroids. Compact spheroids expressed E- and/or P-cadherin, indicating epithelial or epithelial–mesenchymal transition (EMT) hybrid traits, while loose spheroids showed vimentin expression linked to a mesenchymal phenotype. Overall, EMT status, rather than mo-lecular subtype, primarily determined spheroid morphology.