Pain in endometriosis involves not only nociceptive but also neuropathic and neurogenic components, reflecting its complex nature. Histamine, a biogenic amine, has emerged as a critical mediator connecting inflammation and nerve sensitization. This study aimed to characterize histamine receptor (HRH1–HRH4) expression, localization, and related inflammatory mediators in peritoneal, deep infiltrating, and ovarian endometriosis. Gene expression datasets were analyzed, and immunofluorescence staining of endometriotic lesions was performed using immune and neuronal markers. Histamine and its metabolite methylhistamine were quantified in serum, peritoneal fluid, and urine samples. HDC expression was significantly elevated in all endometriotic lesions compared with controls (all p < 0.01), paralleling increased IL-6, COX-2, NGF, and NGFR levels (p < 0.0001). In contrast, HRH1–HRH4 transcript levels showed no significant differences between groups. Immunofluorescence demonstrated robust HRH1–HRH4 protein expression in epithelial, immune, and nerve fibers, with subtype-specific colocalization patterns. Serum histamine concentrations were significantly higher in endometriosis patients than controls (0.484 vs. 0.153 ng/mg protein; p = 0.0014), whereas peritoneal histamine and urinary methylhistamine showed no group differences. Overall, these findings highlight histamine signaling as a potentially important component of endometriosis pathophysiology and point toward new directions for mechanistic studies and therapeutic exploration.

Objective: The late-stage diagnosis and the aggressiveness of high-grade serous tubo-ovarian carcinoma (HGSC) often results in poor survival outcomes, yet some patients exhibit an exceptionally long survival rate.This study aimed to identify molecular profiles associated with long-term/short-term survival in HGSC, with the goal of better understanding protective factors and developing new treatments. Experimental Design: To discover molecular drivers causing aggressiveness of HGSC, tumor samples from 12 long-term HGSC survivors (> 7 years overall survival) and 12 short-term survivors (< 1 year overall survival) were analyzed using targeted RNA sequencing followed by computational analysis. We investigated differentially expressed genes and their functional relevance, inferred differences in cell-type composition and signaling pathways as well as mutation status. To validate our findings, we simulated our study design by using HGSC TCGA dataset samples. We evaluated differential patterns of gene expression between these two groups and developed molecular profiles of HGSC that correlate with survival phenotypes. Results: Besides known molecular cancer drivers and indicators of poor prognosis, we identified specific transcriptional changes between short-term and long-term survivors of HGSC which indicate that immune processes play a fundamental role in long-term survivors. Our computational analysis reveals an important role of the ensemble of IFN-γ signaling and the RFX transcription factors as well as the immune cell composition of the tumor microenvironment. Conclusion: Specific immunologic requirements involving IFN-γ signaling and affected pathways seem to be relevant for long term survival in the generally considered non-immunogenic HGSC, requesting further research to improve diagnostic strategies and targeted therapies.

Head and neck squamous cell carcinoma (HNSCC) with discordant diagnostic patterns of HPV⁺/p16⁻ or HPV⁻/p16⁺ correlate with worse prognosis. This study aims to identify truly HPV-driven HNSCCs using a QuantiGene-Molecular-Profiling-Histology (QG-MPH) assay for identifying transcriptionally active HPV. Of 97 FFPE samples analyzed, 68 were valid for HPV DNA detection by PCR and quantification of HPV E7 and p16 ^INK4a mRNA by QG-MPH. p16 ^INK4a mRNA expression was compared with p16 protein expression via immunohistochemistry (p16 IHC). Among the 68 cases, 26 (38.2%) showed increased high-risk HPV E7 mRNA expression (hrHPV E7 mRNA⁺), while 37 (54.4%) were hrHPV DNA⁺. Concordance between HPV DNA and mRNA status was 70.1%. Notably, 79.2% of E7 mRNA⁺ cases were p16 IHC⁺, compared to 55.9% of DNA⁺ cases, demonstrating better concordance between HPV E7 mRNA⁺ status and p16 mRNA expression plus p16 IHC positivity. All patients (19/19) in the HPV E7 mRNA⁺/p16 IHC⁺ group survived the 5-year follow-up, compared to 59.5% (22/37) in the HPV E7 mRNA⁻/p16 IHC⁻ group (p = 0.001). Specifically, the OS rate was 57.1% (8/14) in the group with discordant HPV DNA and p16 IHC results, compared to 40% (3/5) in the group with discordant HPV E7 mRNA and p16 IHC results. These findings highlight the better outcomes for the transcriptionally active HPV cases and indicate the prognostic disadvantage for patients with discordant patterns and the advantages for incorporating the molecular mRNA profiling by QG-MPH to p16 IHC. In conclusion, QG-MPH quantification of E7 and p16 ^INK4a mRNA more precisely identifies truly HPV-driven from non-HPV-driven HNSCC, compared to HPV DNA testing alone or with p16 IHC, which reduces misclassification and provides valuable implications for improved prognosis prediction and therapeutic decision-making.

Endometriosis (EM) is a chronic inflammatory disease characterized by the growth of endometrial-like tissue outside the uterus, yet its molecular mechanisms remain poorly understood. This study investigated the expression of mucins (MUC1, MUC2, MUC5AC, MUC6, MUC16) and their O-glycans in endometriotic lesions, given their roles in epithelial protection, adhesion, and immune modulation. Using immunohistochemistry, Western blotting, lectin profiling, histochemical staining, and transcriptomic analysis, we compared mucin levels and glycosylation patterns in eutopic and ectopic tissues from women with and without endometriosis and measured mucin-derived tumor markers in serum (CA 125/MUC16 and CA 15-3/MUC1) and peritoneal fluid (CA 125/MUC16). The results showed significant upregulation of all mucins in EM biopsies, with increased MUC1 transcript levels, while MUC6 and MUC16 protein levels did not always align with transcripts. Yet, tumor markers CA 125 and CA 15-3 showed no significant differences between groups. Looking at mucin distribution in biopsies of peritoneal (pEM), deep infiltrating and ovarian EM, MUC1 was significantly overexpressed in lesions of all EM forms, while MUC5AC was significantly elevated in pEM. Lectin analysis revealed specific glycan changes, including elevated core-1 O-glycans and α(1-2)-linked fucosylation, while sialylation remained unchanged. These findings demonstrate consistent mucin dysregulation and glycan alterations, implicating their roles in epithelial adhesion, immune evasion, and lesion persistence. Mucin biology thus emerges as a promising target for diagnostic and therapeutic strategies in endometriosis.

Figure S9 shows CIBERSORT estimated celltypes per sample in the Charité cohort and in the TCGA cohort.

Figure S7 shows the Mutation Landscape of TCGA LTS and STS Patients.