Distinct genetic alterations and luminal molecular subtype in nested variant of urothelial carcinoma (NVUC)

AIMS: Nested variant of urothelial carcinoma (NVUC) is rare and only few small series exist. Molecular characteristics and the classifying marker profile as well as therapeutic targets of this specific variant are mostly unknown. Aim of this study was to characterize NVUC on the molecular level in one of the largest cohorts to date. In addition, we applied an immunohistochemical marker panel in order to define the molecular subtype. METHODS: 60 NVUC cases were collected from different departments. TERT mutation analysis was carried out in all samples using SNaPshot analysis. Target sequencing of 48 cancer related genes by Next Generation Sequencing (NGS) analysis was performed in a subset of 26 cases. Immunohistochemical markers CD44, CK5, CK14, EGFR, p63, FOXA1, GATA3, CD24 und CK20 were used to elucidate the molecular subtype. RESULTS: A total of 62.5% of NVUC cases harbored a mutation of the TERT promoter. Additionally, TP53, JAK3, and CTNNB1 were among the most frequently mutated genes identified by NGS analysis. Subtyping revealed that all NVUC express luminal markers such as CD24, FOXA1, GATA3 and CK20. CONCLUSIONS: Summarized, NVUC belong to the luminal molecular subtype. Moreover, a subset of NVUC seems to be characterized by mutations of the Wnt- and inflammatory pathway, including JAK3 mutations, indicating a different biological background compared to conventional urothelial bladder cancer.