Abhishek Tripathi

Demographic and clinical characteristics of the study cohort (N = 30).

Immunohistochemical expression for 21 cell surface markers across testicular germ cell tumor subtypes and histologic components.

Immunohistochemical staining of selected cell surface markers across testicular GCT subtypes. A, CLDN6 (membranous staining) in seminoma, (B) CLDN6 (membranous staining) in yolk sac tumor, (C) EGFR (membranous staining) in seminoma, (D) EGFR (membranous staining) in teratoma, (E) TAG72 (membranous and cytoplasmic staining) in seminoma, (F) TAG72 (membranous and cytoplasmic staining) in choriocarcinoma, (G) TROP2 (membranous staining) in seminoma, (H) TROP2 (membranous staining) in teratoma, (I) Nectin-4 (membranous and cytoplasmic staining) in seminoma, (J) Nectin-4 (membranous and cytoplasmic staining) in teratoma, (K) GPNMB (membranous and cytoplasmic staining) in seminoma, (L) GPNMB (membranous and cytoplasmic staining) in embryonal carcinoma, (M) c-MET (nuclear staining) in seminoma, (N) c-MET (nuclear staining) in embryonal carcinoma, (O) HER2 (membranous staining) in teratoma, and (P) c-MET (membranous and cytoplasmic staining) in choriocarcinoma.

Antibodies used for immunohistochemistry, including clone, catalog number, and vendor.

To characterize clinically actionable cell surface proteins in testicular germ cell tumors (GCT), we retrospectively analyzed archival formalin-fixed, paraffin-embedded orchiectomy specimens from 30 patients, including 11 with pure seminoma and 19 with mixed GCTs. Immunohistochemistry was performed for 21 surface antigens using an automated platform. Staining intensity (0 to 3+) and the percentage of positive tumor cells were used to derive H-scores (0–300). Expression was summarized by patient classification and histologic component; high expression was defined as H-score ≥100. Claudin-6 (CLDN6) was diffusely and strongly expressed, with H-score ≥100 in 100% of pure seminomas and 89.5% of mixed GCTs. At the component level, 100% of seminoma and 92.9% of embryonal carcinoma samples reached this threshold. Epidermal growth factor receptor (EGFR) showed high expression in 68.4% of mixed GCTs and 45.5% of pure seminomas, with most enrichment in teratoma components (73.3%, ≥100; mean H-score, 212). TROP2 was markedly upregulated in mixed GCTs (78.9%, ≥100; mean, 194.7), particularly in teratoma components (86.7%, ≥100). Nectin-4 and glycoprotein nonmetastatic melanoma B (GPNMB) were expressed in smaller but notable subsets, including teratoma and seminoma, whereas most remaining antigens, including CD19, DLL3, FOLR1, and BCMA, showed minimal or no expression. These findings demonstrate a distinct surface antigen landscape in testicular GCTs, with CLDN6, EGFR, and TROP2 emerging as leading candidates for further translational evaluation. The enrichment of these targets highlights antigenic heterogeneity within GCTs and supports further validation in appropriate disease settings, including metastatic, posttreatment, and relapsed or refractory disease.Significance:Relapsed or refractory testicular GCT remains a clinical challenge with limited treatment options. We evaluated cell surface antigens of primary GCTs and identified CLDN6, EGFR, and TROP2 as candidate targets for antigen-directed approaches. These findings provide a rationale for future validation in metastatic and posttreatment disease settings.

Background: Sarcomatoid carcinoma of the upper urinary tract (SCUT) is a rare and aggressive malignancy. Due to its rarity, the molecular landscape and the prevalence of potentially targetable alterations are poorly characterized. We aimed to compare the clinical, pathological, and genomic profiles of SCUT and upper tract urothelial carcinoma (UTUC). Methods: We leveraged the Tempus Lens clinically annotated genomic dataset to extract clinicopathologic and somatic genomic alteration data from patients with UTUC and SCUT. Patients with any-stage disease who underwent either blood- or tissue-based next-generation sequencing were included. Baseline clinical and demographic characteristics were summarized using descriptive statistics. Comparisons between groups were performed using Wilcoxon rank-sum test for continuous variables and the Chi-square test/Fisher’s exact test for categorical variables. Mutational frequencies and pairwise comparisons were performed to assess significant differences between groups. Results: In total, 1721 patients were included, of which 1600 (93%) had UTUC and 121 (7%) had SCUT. Patients with SCUT were younger at diagnosis, 61 years (interquartile range (IQR) 54, 70), compared to UTUC, 71 years (IQR 64, 77) (p < 0.001), and were more likely to have node-positive disease at presentation (all p < 0.001). SCUT patients were more likely to show visceral metastases to the lung (44% vs. 21%), bone (31% vs. 17%), and brain (7% vs. 1%), compared to UTUC (all p < 0.05). Among SCUT patients, the most common genomic alterations were TERT (30%), TP53 (29%), NF2 (19%), PTEN (13%), SETD2 (12%), PBRM1 (12%), and BAP1 (8%). Among UTUC patients, the most common were TERT (52%), TP53 (52%), KMT2D (30%), FGFR3 (25%), ARID1A (20%), and KDM6A (18%). Compared to UTUC, SCUT was significantly enriched with NF2, SETD2, PBRM1, PTEN, and BAP1 (all p < 0.05). SCUT was depleted in FGFR3 (0% vs. 25%) and FGF4 (0% vs. 8%) mutations compared to UTUC (both p < 0.05). Targetable alterations were observed in SCUT, including NF2, SETD2, and PTEN. Conclusion: Compared with UTUC, SCUT exhibits a more aggressive clinical and genomic phenotype, characterized by enrichment in NF2, SETD2, PBRM1, and PTEN. These findings underscore the divergent molecular landscape of SCUT and highlight potentially targetable genomic alterations.

To characterize clinically actionable cell surface proteins in testicular germ cell tumors (GCTs), we retrospectively analyzed archival formalin-fixed paraffin-embedded orchiectomy specimens from 30 patients, including 11 with pure seminoma and 19 with mixed GCTs. Immunohistochemistry was performed for 21 surface antigens using an automated platform. Staining intensity (0-3+) and percentage of positive tumor cells were used to derive H-scores (0-300). Expression was summarized by patient classification and histologic component; high expression was defined as H-score ≥100. CLDN6 was diffusely and strongly expressed, with H-score ≥100 in 100% of pure seminomas and 89.5% of mixed GCTs. At the component level, 100% of seminoma and 92.9% of embryonal carcinoma samples reached this threshold. EGFR showed high expression in 68.4% of mixed GCTs and 45.5% of pure seminomas, with most enrichment in teratoma components (73.3% ≥100; mean H-score 212.0). TROP2 was markedly upregulated in mixed GCTs (78.9% ≥100; mean 194.7), particularly in teratoma components (86.7% ≥100). Nectin-4 and GPNMB were expressed in smaller but notable subsets, including teratoma and seminoma, whereas most remaining antigens, including CD19, DLL3, FOLR1, and BCMA, showed minimal or no expression. These findings demonstrate a distinct surface antigen landscape in testicular GCTs, with CLDN6, EGFR, and TROP2 emerging as leading candidates for further translational evaluation. Enrichment of these targets highlights antigenic heterogeneity within GCTs and supports further validation in appropriate disease settings including metastatic, post-treatment, and relapsed or refractory disease.

Supplementary File 1 from Randomized Phase II Study Evaluating the Addition of Pembrolizumab to Radium-223 in Metastatic Castration-resistant Prostate Cancer

Clinical characteristics at time of randomization of PSA responders vs. non-responders

Immune cell functional states differ in clinical responders compared to non-responders. (A) Heat maps summarizing log2 fold changes resulting from statistical scaffold analysis of functional markers 4-1BB, CD44, CTLA-4, GITR, HLA-DR, ICOS, Ki-67, OX40, PD-1, PD-L1, PD-L2, TIGIT, TIM3 and VISTA are shown for Week 1, Week 5, and Week 9, when comparing responders to non-responders as defined by rPFS.