Despite many treatment strategies available for metastatic renal cell carcinoma (mRCC), predictive biomarkers of response to immunotherapy are still needed. In this context, growing efforts have been devoted to translational research, especially focusing on the immune tumor microenvironment (I-TME). The Meet-URO 18 is a multicentric retrospective study assessing the I-TME of mRCC patients receiving ≥ 2nd line nivolumab, divided into responders and non-responders according to clinical benefit [progression-free survival ≥ 12 and ≤ 3 months]. The primary objective was to identify differential immunohistochemical and molecular patterns between the two groups. We present the transcriptomic analysis performed on primary tumor tissues using a custom NanoString panel of 66 genes from the D’Costa et al. signature, grouped into angiogenesis, T-effector response, tumor invasion, and calcium signaling. Forty-two samples (25 responders and 17 non-responders) underwent transcriptomic analysis using the NanoString 66-gene immune-oncology panel. Hierarchical clustering recapitulated the transcriptional axes described by D’Costa et al. but did not distinguish patients by immunotherapy response. No genes were significantly differentially expressed after multiple testing correction; however, CD34 showed the strongest nominal association with responder tumors and was upregulated in this group, but did not retain statistical significance after adjustment. CD34 expression also correlated with CD8+ T-cell infiltration. Our findings confirm the applicability of the D’Costa molecular signature and identify CD34 as the strongest nominally associated transcript in responder tumors, warranting further orthogonal and prospective validation.

Introduction: Most rectal cancers are microsatellite-stable (MSS) and derive limited benefit from immune checkpoint inhibition. We assessed whether familial colorectal cancer aggregation defines a distinct biological context within MSS rectal cancer.Materials and Methods: We performed a prespecified analysis of two prospective multicentre cohorts conducted between 2018 and 2024. Patients with MSS rectal adenocarcinoma were classified as familial (FH⁺) or sporadic (FH⁻), with a predefined subgroup of patients with an affected first-degree relative (FDR⁺). Known hereditary colorectal cancer syndromes were excluded. Immune profiling, transcriptomic analysis and targeted sequencing were performed on tumour-adjacent histologically normal rectal mucosa and, when appropriate, tumour tissue. Analyses were stratified by neoadjuvant treatment status.Results: Among 374 patients, 92 were FH⁺ and 65 were FDR⁺. In NAT-naïve patients, FDR⁺ cases showed higher epithelial CD80⁺ cell density than FH⁻ cases. After neoadjuvant therapy, FH⁺ patients had increased activated CD8⁺CD28⁺ T cells and reduced epithelial HLA-ABC expression, whereas FDR⁺ cases showed lower CD3⁺ T-cell density. Transcriptomic analysis indicated a quiescent mucosal phenotype in FH⁺ patients, with reduced DNA repair, proliferative, metabolic, angiogenic, and immune pathway activity. Post-NAT FH⁺ and FDR⁺ patients showed higher mutational ratios. In FH⁻, but not FH⁺, higher mutational ratio was associated with improved disease-free survival and immune activation.Discussion: Familial aggregation may define a biologically distinct MSS rectal cancer subgroup, characterised by altered epithelial–immune coordination and dissociation between therapy-induced genomic stress and immune surveillance.​

Objective: Sporadic medullary thyroid carcinoma (sMTC) is predominantly driven by somatic RET or RAS mutations, although the molecular basis of disease heterogeneity remains incompletely understood. Our study aims to characterize molecular heterogeneity in sMTC in order to identify pathways that may improve patient’s stratification and support more personalized clinical management strategies. Methods: Deep targeted next-generation sequencing of 31 neuroendocrine- and cancer-related genes was performed on tumor samples from 94 patients with sMTC to characterize the somatic mutational landscape beyond canonical drivers. Results: RET and RAS mutations were detected in 53% and 29% of cases, respectively, while 18% of tumors lacked both alterations. Variant allele frequency (VAF) analysis demonstrated a significant positive correlation between driver mutation burden and tumor size in both RET- and RAS-mutant tumors, supporting the clonal contribution of these alterations. Additional oncogenic variants were identified in genes involved in DNA damage response and epigenetic regulation, including ATM and KMT2A. Notably, within the RAS-mutant subgroup, the presence of co-occurring oncogenic alterations was associated with more advanced T status (T3-T4, p = 0.0181) at diagnosis and lower biochemical cure rates (p = 0.02) at the follow-up compared with tumors harboring isolated RAS mutations, supporting the clinical relevance of extended genomic profiling in RAS-mutant sMTC. Conclusions: Overall, these findings highlight additional oncogenic alterations potentially involved in tumor progression and suggest that extended targeted profiling may provide clinically relevant information on molecular heterogeneity in sMTC, particularly within RAS-mutant tumors.

Background Thyroid nodules are rare in children but carry a markedly higher risk of malignancy compared to adults (20%-26% vs. 5%). Hyperfunctioning thyroid nodules are exceptionally uncommon in the pediatric population and are typically benign. We describe a rare case of a hyperfunctioning thyroid nodule in a prepubertal child that was ultimately diagnosed as an angioinvasive encapsulated follicular carcinoma, arising in the context of the WHO entity 'follicular adenoma showing papillary architecture'. In addition, we provide a comprehensive review of published pediatric cases of hyperfunctioning thyroid nodules with malignant histology.Case Presentation A 12-year-old boy presented with a left-sided thyroid nodule detected on ultrasound after cervical swelling. Laboratory evaluation revealed suppressed thyroid-stimulating hormone (TSH), with free thyroxine (fT4) and free triiodothyronine (fT3) within the reference range. Serial ultrasound examinations revealed a progressively enlarging mildly hypoechoic nodule, autonomously functioning on thyroid scintigraphy. Fine-needle aspiration cytology (FNAC) was classified as TIR2. Surgical excision showed a follicular-patterned neoplasm showing papillary features, focal capsular and vascular invasion and no lymph node metastases. Comprehensive molecular analysis identified a pathogenic somatic variant in the GNAS gene, whereas no alterations were detected in TSHR, BRAF, RAS genes, the TERT promoter, or DICER1.Conclusion This case highlights the importance of an integrated evaluation in pediatric thyroid nodules, particularly when hyperfunction coexists with indeterminate or suspicious imaging features. Follicular-patterned thyroid neoplasms showing papillary architecture pose diagnostic challenges in children, and molecular analysis may assist risk stratification and elucidate pathogenetic mechanisms linking autonomous function and malignant transformation.

Background: Anastomotic leaks (ALs) remain a critical complication after rectal cancer surgery. Emerging evidence suggests that local immune dysregulation may play a key role in anastomotic healing. We investigated the immune microenvironment of histologically normal, tumor-adjacent rectal mucosa-a tumor-conditioned field-as a potential substrate for AL predisposition. Methods: IMMUNOREACT 4 is a sub-analysis of the IMMUNOREACT project (clinicaltrials.gov NCT04915326 and NCT04915326), a multicenter translational study evaluating immune features of histologically normal, tumor-adjacent rectal mucosa of patients undergoing colorectal anastomosis. A prospective cohort (n = 121) was analyzed using flow cytometry, in addition to a retrospective cohort (n = 262) using immunohistochemistry. Immune markers of epithelial activation and lymphocyte subsets were compared between patients with and without postoperative ALs. Exploratory predictive models combining immune and clinical variables were developed and evaluated using discrimination, calibration and decision curve analyses. Results: At flow cytometry, the CK+HLAabc+ MFI (AUC 0.66, 95% CI 0.52-0.80), CD8+CD38+ cell rate (AUC 0.65, 95% CI 0.52-0.78) and CD3+CTLA4+ cell rate (AUC 0.65, 95% CI 0.51-0.80) showed moderate predictive potential for ALs. In immunohistochemistry, CD3+ (AUC 0.57, 95% CI 0.54-0.60), CD8+ (AUC 0.57, 95% CI 0.52-0.62), CD8β+ (AUC 0.59, 95% CI 0.53-0.65) and Tbet+ (AUC 0.60, 95% CI 0.56-0.64) showed some predictive ability for ALs. The model including CD8β+, the BMI, neutrophile/lymphocyte ratio and tumor location had an AUC of 0.67 (95% CI 0.62-0.72). Conclusions: Immune activation within histologically normal, tumor-adjacent rectal mucosa-characterized by epithelial HLA upregulation and cytotoxic or Th1 T cell infiltration-is associated with postoperative ALs. Although predictive accuracy is limited, these findings support the concept that a tumor-conditioned immune microenvironment may predispose patients to impaired anastomotic healing. Integration of mucosal immune profiling with clinical variables represents a promising exploratory approach that warrants further prospective validation.

Esophageal cancer, particularly squamous cell carcinoma (SCC) and adenocarcinoma (EAC), is a major contributor to cancer-related mortality. The different histopathologic subtypes have different pathological origins, epidemiology and prognosis. TNM staging system allows to stratify the prognosis and determine the most appropriate treatment. Surgery remains the gold standard for treating early-stage esophageal cancer, including various procedures that can adapt to the singular cases helping to reduce morbidity. However, esophagectomy remains burdened by considerable postoperative complications, such as anastomotic leakage, pleural effusion, pneumonia, acute respiratory distress syndrome, stricture formation, chylothorax, delayed gastric emptying, hiatal herniation, and reflux esophagitis. Accurate radiologic evaluation plays a crucial role in detecting, characterizing, and staging esophageal cancer, directly influencing treatment strategies. Radiological imaging is pivotal in determining patient's prognosis, both for the management of post-operative complications and for long-term follow-up. A thorough understanding of the imaging characteristics and underlying pathology is essential for improving diagnostic accuracy and guiding therapeutic decision-making.

HYADD®4 is a hexadecylamide derivative of Hyaluronan (HA) forming hydrogels with excellent lubricating and viscoelastic properties, widely used as viscosupplement in the treatment of knee OA. In this study, the effects of the intra-articular (i.a.) injection of FID-337, a ready-to-use formulation of HYADD®4 + Alendronate (ALN), and FID-338, a combined treatment based on HYADD®4 + Rapamycin (RAP), were evaluated on an OA model obtained by destabilization of the medial meniscus (DMM) performed in ovariectomized (OVX) female rats. Eight-week-old female Sprague Dawley rats were ovariectomized and after four weeks underwent DMM surgery on the right knee. Four weeks after, the animals were randomly divided into 4 groups of 9 rats each according to knee joint i.a. treatment, administered every 2 weeks (a total of 3 injections): FID-337, FID-338, HYADD®4 and saline; sham-operated rats (SHAM) were used as control. Mechanical allodynia was assessed weekly by the Von Frey test. Four weeks after the third i.a. injection rats were sacrificed. Microtomography (micro-CT) analysis was performed to assess bone volume, trabecular thickness and separation, and bone mineral density. Hematoxylin–eosin and safranin O/fast green staining were used to evaluate cartilage degradation, which was quantified through the OARSI score, and proteoglycan (PG) content. Withdrawal thresholds, which were low and unchanged over time in rats treated with saline, increased after the first injection of each treatment, reached levels of the SHAM group by the 10th week in the FID-338 group while continued to increase until the end of the study in the FID-337 group. The bone architecture was altered in NaCl-treated rats, and partially restored only after treatment with FID-337, FID-338 or HYADDÒ4. In the same groups, the synovial membrane was better preserved compared to the saline group. The OARSI (Fig.1) and PG loss scores were significantly lower in HYADD®4-treated joints and even further reduced significantly with FID-337 or FID-338 treatment. I.a. administration of HYADD®4 with ALN or RAP significantly impacts the OA progression, reducing pain and preventing cartilage damage in the OVX+DMM rat model, which describes a specific OA endotype. The local administration could maximize the local benefit while reducing the well known systemic side effects and this may lead to the development of a novel therapy with repurposed drugs combined with the HA-based viscosupplement. For any figures or tables, please contact the authors directly.

AIMS:Current understanding of the risk of neoplastic progression in patients with Barrett's esophagus with indefinite for dysplasia (BE-IND) and gastric indefinite for dysplasia (G-IND) remains limited. This study aims to identify prognostic histological and clinicopathological factors for IND progression in the upper gastrointestinal tract. METHODS AND RESULTS:Patients with confirmed BE-IND and G-IND, no previous evidence of dysplasia/cancer and follow-up of ≥ 6 months were included. The rate of neoplastic progression was calculated and the multivariate Cox regression model adjusted for demographic and histological features was used to identify risk factors for progression. A total of 719 patients diagnosed with IND (158 BE-IND and 561 G-IND) were identified, 395 of whom were excluded. Progression rates were 4.4 per 100 person-year for BE-IND and 1.6 per 100 person-year for G-IND patients. Progression was observed only in IND of hyperproliferative intestinal metaplasia (HIM) type. Operative link for gastritis assessment (OLGA) stage (III-IV versus 0-II) was the only significant predictor of G-IND progression [hazard ratio (HR) = 47.82, confidence interval (CI) = 5%: 6.24-366.37, P < 0.001]. In BE-IND patients, lack of interval endoscopy significantly increased the risk for BE-IND (HR = 13.45, CI = 95%: 1.24-145.75, P = 0.032). CONCLUSION:IND is a challenging diagnosis for pathologists and implies an increased risk for neoplasia, especially in BE patients, without providing definitive information for patient management. This study highlights that neoplastic progression of IND lesions is primarily associated with HIM type. For BE-IND, interval endoscopy significantly reduces progression risk, while in G-IND, OLGA staging (III-IV) is a strong predictor of progression. These findings emphasise the importance of identifying HIM and using OLGA staging in G-IND for better risk stratification and follow-up strategies.

CONTEXT:The search for somatic mutations in adrenals resected from patients with primary aldosteronism (PA) is performed by Sanger sequencing, often implemented with immunohistochemistry (IHC)-guidance focused on aldosterone-producing (CYP11B2-positive) areas. OBJECTIVE:To investigate the impact of double IHC for CYP11B1 and CYP11B2 on Sanger and next-generation sequencing (NGS). METHODS:We investigated 127 consecutive adrenal aldosterone-producing adenomas from consenting surgically cured PA patients using double IHC for CYP11B1 and CYP11B2, by Sanger sequencing and NGS. RESULTS:Double IHC for CYP11B2 and CYP11B1 revealed 3 distinct patterns: CYP11B2-positive adenoma (pattern 1), mixed CYP11B1/CYP11B2-positive adenoma (pattern 2), and adrenals with multiple small CYP11B2-positive nodules (pattern 3). Sanger sequencing allowed detection of KCNJ5 mutations in 44% of the adrenals; NGS revealed such mutations in 10% of those negative at Sanger and additional mutations in 61% of the cases. Importantly the rate of KCNJ5 mutations differed across patterns: 17.8% in pattern 1, 71.4% in pattern 2, and 10.7% in pattern 3 (χ2 = 22.492, P < .001). CONCLUSION:NGS allowed detection of mutations in many adrenals that tested negative at Sanger sequencing. Moreover, the different distribution of KCNJ5 mutations across IHC patterns indicates that IHC-guided sequencing protocols selecting CYP11B2-positive areas could furnish results that might not be representative of the entire mutational status of the excised adrenal, which is important at a time when KCNJ5 mutations are suggested to drive management of patients with aldosterone-producing adenomas.

Although hypercoagulability is commonly associated with malignancies, whether coagulation factors directly affect tumor cell proliferation remains unclear. Herein, by performing single-cell RNA sequencing (scRNA-seq) of the prostate tumor microenvironment (TME) of mouse models of castration-resistant prostate cancer (CRPC), we report that immunosuppressive neutrophils (PMN-MDSCs) are a key extra-hepatic source of coagulation factor X (FX). FX activation within the TME enhances androgen-independent tumor growth by activating the protease-activated receptor 2 (PAR2) and the phosphorylation of ERK1/2 in tumor cells. Genetic and pharmacological inhibition of factor Xa (FXa) antagonizes the oncogenic activity of PMN-MDSCs, reduces tumor progression, and synergizes with enzalutamide therapy. Intriguingly, F10high PMN-MDSCs express the surface marker CD84 and CD84 ligation enhances F10 expression. Elevated levels of FX, CD84, and PAR2 in prostate tumors associate with worse survival in CRPC patients. This study provides evidence that FXa directly promotes cancer and highlights additional targets for PMN-MDSCs for cancer therapies.

Introduction: The Meet-URO 18 study is a multicentric study of patients with metastatic renal cell carcinoma receiving nivolumab in the second-line and beyond, categorized as responders (progression-free survival ≥ 12 months) and non-responders (progression-free survival < 3 months). Areas covered: The current study includes extensive immunohistochemical analysis of T-lineage markers (CD3, CD4, CD8, CD8/CD4 ratio), macrophages (CD68), ph-mTOR, CD15 and CD56 expression on tumor cells, and PD-L1 expression, on an increased sample size including 161 tumor samples (113 patients) compared with preliminary presented data. Responders' tumor tissue (n = 90; 55.9%) was associated with lower CD4 expression (p = 0.014), higher CD56 expression (p = 0.046) and higher CD8/CD4 ratio (p = 0.030). Expert opinion/commentary: The present work suggests the regulatory role of a subpopulation of T cells on antitumor response and identifies CD56 as a putative biomarker of immunotherapy efficacy.

BACKGROUND:Congenital adrenal hyperplasia (CAH) encompassed a bunch of autosomal recessive disorders characterized by impaired cortisol levels due to an enzymatic deficiency in steroid synthesis. In adult male patients with CAH, a frequent complication related to poor disease control is the development of ectopic adrenocortical tissue in the testes, named testicular adrenal rest tumors (TART). Conversely, ovarian adrenal rest tumors (OART) in females are extremely rare and adrenal rests in sites other than gonads are so uncommon to have been described only few times in literature. CASE PRESENTATION:We report a case of a male patient with untreated CAH and oncologic history of pleomorphic sarcoma who presented with massive bilateral adrenal enlargement and adrenal rest tumors in peri-lumbar and peri-cecal sites, which mimicked metastasis from sarcoma. CONCLUSIONS:The development of massive adrenal enlargement and ectopic adrenal rest tumors in sites other than gonads, even if very uncommon, should be suspected in patients with CAH and prolonged periods of undertreatment.

Aldosterone (Aldo) exerts its action through binding with the mineralocorticoid receptor (MR). Clinically, a link between primary aldosteronism (PA) and thyroid diseases has been hypothesised. However, the presence and activity of MR on the thyroid have not yet been demonstrated. We investigated the gene/protein expression and activation of MR in primary thyroid cell cultures (normal rat thyroid [FRTL-5] and human papillary thyroid cancer [PTC] cell lines, BCPAP and K1) through qRT-PCR analysis, immunofluorescence, and confocal microscopy. We also studied the effects of Aldo on thyroid-specific and inflammation genes in vitro. Paired human normal and neoplastic thyroid tissues were also studied. We demonstrated both gene and protein expression and activation of MR in normal rat thyroid and human PTC lines. Incubation with Aldo induced an acute increase in IL-6 expression in both the FRTL-5 and BCPAP lines, which was antagonised by spironolactone, and an acute and late upregulation of thyroid-specific genes in FRTL-5. MR was also expressed at both gene and protein levels in normal human thyroid tissues and in PTC, with a progressive decline during neoplastic tumourigenesis, particularly in more aggressive histotypes. We present the first evidence of MR gene and protein expression in both normal and pathological thyroid cells and tissues. We have shown that MR is present and functionally activated in thyroid tissue. Binding of Aldo to MR induces the expression of inflammatory and thyroid-specific genes, and the thyroid may thus be considered a novel mineralocorticoid target tissue.