Purpose Accurate prediction of lymph node metastasis (LNM) remains challenging in early gastric cancer (EGC), particularly when determining the need for additional surgery after endoscopic resection. We developed and evaluated a deep learning framework using multiple pathology foundation models for LNM prediction and explored histological features associated with model-predicted metastatic risk. Materials and Methods Foundation model ensembles were developed using whole-slide images from surgical EGC cases and evaluated in internal and external surgical cohorts. The model was further tested in an endoscopic submucosal dissection cohort in which all patients subsequently underwent additional gastrectomy with lymph node dissection. Performance was assessed by area under the curve (AUC) and compared with scratch-trained baseline models. At a fixed sensitivity of 1.00, the potential reduction in lymph node dissection among histologically node-negative patients was evaluated. Model-guided histological analysis was performed to interpret features associated with predicted metastatic risk. Results The final ensemble model achieved an AUC of 0.879 (95% CI, 0.799-0.944) in the internal and 0.801 (95% CI, 0.697-0.891) in the external cohort, representing improvements of 0.040 and 0.066, respectively, over the baseline model. In the endoscopic submucosal dissection cohort, the model achieved an AUC of 0.766 (95% CI, 0.635-0.882), representing an improvement of 0.189 over the baseline model. At a fixed sensitivity of 1.00, the model suggested avoiding lymph node dissection in approximately 5% of histologically node-negative patients. Model-guided histological analysis showed associations consistent with established pathological features, with exploratory observations including context-dependent associations between inflammatory infiltration and tumor differentiation, differences in predicted risk between well- and moderately differentiated tumors, and lower predicted risk in poorly cohesive carcinoma. Conclusions A multiple foundation model ensemble showed improved LNM risk prediction in EGC and provided interpretable histological insights supporting clinical understanding and risk stratification.
Purpose Consensus molecular classes of urothelial carcinoma (UC) include basal/squamous (Ba/Sq), luminal papillary (LumP), luminal nonspecified (LumNS), luminal unstable (LumU), stroma-rich, and neuroendocrine (NE)-like. We aimed to determine the consensus molecular classifications of micropapillary (MP), plasmacytoid (PC), and sarcomatoid (SM) subtypes of bladder UC and characterize their spatial transcriptomic profiles. Materials and Methods A tissue microarray of 17 FFPE cores (5 MP, 6 PC, 6 SM) was profiled for expression of 18,676 mRNAs using the Bruker GeoMx Digital Spatial Profiler. 38 areas of interest (AOIs) were selected based on H&E and fluorescence markers. Each AOI was assigned to a consensus molecular class using a published approach, and pathway analysis was performed using Reactome gene sets. Results Most MP and PC demonstrated luminal molecular classes, whereas SM was predominantly Ba/Sq. Adjacent mixed histological components within the same tumor frequently corresponded to distinct consensus molecular classes. Ba/Sq tumors upregulated invasion- and extracellular matrix (ECM) remodeling-associated genes, whereas luminal classes demonstrated distinct differentiation-associated signatures. MP demonstrated a predominantly luminal epithelial transcriptional program characterized by increased expression of luminal/urothelial differentiation markers (KRT20, UPK1A, PSCA, FOXA1), secretory-associated genes (LCN2, MUC1, MUC20, SPINK1), and relative ERBB2 enrichment, with suppression of basal/squamous-associated genes (KRT5, KRT6A, KRT14, TP63). In contrast, SM demonstrated a predominantly mesenchymal/stromal transcriptional program characterized by enrichment of mesenchymal/ECM-associated genes (FN1, VIM, COL6A1, TGFB1), and reduced expression of luminal/epithelial and adhesion-associated genes (TACSTD2, PSCA, NECTIN4, CDH1). PC appeared to demonstrate an intermediate phenotype between MP and SM, with partial preservation of epithelial differentiation and secretory programs, reduced adhesion programs, and moderate stromal-associated features. The therapeutic target NECTIN4 was relatively enriched in MP and PC compared with SM, whereas FGFR3 was relatively downregulated in MP and PC compared with CIUC. Pathway analysis demonstrated relative enrichment of IGF/IGFBP signaling across MP, PC, and SM, with MP enriched for protein secretory programs, PC for RTK/MAPK and WNT/β-catenin signaling, and SM for ECM remodeling. Conclusions Adjacent histological components frequently corresponded to different consensus molecular classes in UC. MP, PC, and SM demonstrated distinct transcriptomic profiles, with PC exhibiting an intermediate phenotype between MP and SM.
PURPOSE:To assess whether pretreatment intratumoral macrophage-associated and T-cell marker-positive areas are associated with treatment outcomes in patients with stage IV solid tumors receiving programmed cell death protein 1 (PD-1)-based immune checkpoint blockade, and whether tissue marker-positive areas correspond to peripheral blood monocyte subsets. MATERIALS AND METHODS:This retrospective, single-center study included 52 patients with metastatic or recurrent stage IV solid tumors and available pretreatment formalin-fixed, paraffin-embedded tumor tissue. Immunohistochemistry was performed for CD3, CD8, CD14, CD16, CD68, CD86, CD163, and CD206. Automated image analysis quantified chromogenic marker-positive area as a percentage of the analyzed tumor region of interest. Marker-positive areas were compared between patients with objective response, defined as complete or partial response, and those with stable or progressive disease. Associations with progression-free survival and overall survival were evaluated using median-based Kaplan-Meier analyses and Cox proportional hazards models. Correlations with peripheral blood monocyte subsets were assessed in patients with available flow-cytometry data. Exploratory sensitivity analyses were performed in the anti-PD-1 monotherapy subgroup and within major tumor types. RESULTS:Higher CD68-, CD86-, CD163-, CD206-, and CD8-positive areas were associated with longer progression-free survival and overall survival, whereas CD14-, CD16-, and CD3-positive areas were not. All six myeloid/macrophage-associated marker-positive areas and CD8-positive area were higher in patients with complete or partial response than in those with stable or progressive disease. Tissue marker-positive areas did not show consistent correlations with peripheral blood monocyte subsets. The direction of the principal associations was maintained in the anti-PD-1 monotherapy sensitivity analysis. CONCLUSIONS:Higher pretreatment CD68-, CD86-, CD163-, CD206-, and CD8-positive areas were associated with favorable outcomes during PD-1-based immune checkpoint blockade in this heterogeneous stage IV cohort. These exploratory findings do not establish a predictive biomarker or patient-selection test. Prospective validation in larger, tumor-specific cohorts is required.
PURPOSE:Tumor stroma, the cellular and non-cellular support system surrounding cancer cells, is a promising source of novel therapeutic targets. Extradomain-B of fibronectin (EDB+FN), a tumor-specific antigen expressed in the extracellular matrix (ECM) of tumor stroma and minimally in healthy tissues, is the target of the first-in-concept antibody drug conjugate (ADC) micvotabart pelidotin (MICVO) which is currently in clinical trials (NCT05720117 and NCT06795412) for the treatment of solid tumors. A specific and reproducible immunohistochemistry (IHC) assay and scoring approach are needed to detect EDB+FN protein expression for indication selection, preclinical research, and potential future evaluation of correlations between target expression and therapeutic efficacy. MATERIALS AND METHODS:Here, an EDB+FN IHC assay was developed to detect EDB+FN protein in human solid tumors. A novel scoring system was developed adapting H-score methodology to evaluate intensity and localization of EDB+FN extracellularly in tumor stroma and cellularly in cancer cells in ten solid tumor types. Moreover, a custom, AI-powered digital pathology algorithm was developed to streamline scoring of EDB+FN protein expression for research and future analysis of spatial distribution. RESULTS:The developed IHC assay reproducibly detected EDB+FN in the tumor stroma across FFPE samples from ten tumor types and minimally in normal tissue. Percent stroma was not predictive of EDB+FN expression within a tumor, suggesting heterogeneous EDB+FN protein expression in tumor stroma. The modified H scoring approach reproducibly quantified EDB+FN expression extracellularly in tumor stroma by pathologist evaluation and a digital algorithm built with open-source tools demonstrated proof-of-concept for digital quantification of EDB+FN protein expression. CONCLUSIONS:This work demonstrates technical characterization of a research-use IHC assay for detection of EDB+FN in FFPE tumor sections, reveals broad expression of EDB+FN across cancer types, and establishes a novel approach for scoring expression of a non-cell associated protein in the tumor ECM, both by pathologists and using digital pathology.
PURPOSE:Repeated evidence demonstrates limited reproducibility and accuracy of the visual quantification (VQ) of the tumor cell content (TCC) by clinical pathologists for downstream molecular testing. Artificial intelligence (AI)-based digital quantification (DQ) of TCC represents a promising alternative, yet real-world evidence from routine molecular diagnostic workflows remain limited. In this study, we evaluated the analytical performance and practical aspects of analytical validation process of an AI-based DQ tool in routine molecular diagnostics. MATERIAL AND METHODS:The clinical-grade AIM-TumorCellularity (AIM-TC; PathAI©) workflow was tested in molecular diagnostics for samples analyzed by comprehensive genomic profiling (FoundationOne®CDx (F1CDx), Foundation medicine Inc.). The cohort included 300 non-paired resection, biopsy, and cytology/cell block) specimens from primary and metastatic breast (n = 66), lung (n = 117), colorectal (n = 40), pancreatic (n = 38), and prostate (n = 39) cancers, reflecting real-world diagnostic sample heterogeneity of a tertiary care center. We compared TCC estimates generated by pathologists' VQ, AI-based DQ, and molecular quantification (MQ) by bioinformatic deconvolution. RESULTS:Agreement was lowest between VQ and MQ (Spearman Rs = 0.38) and between VQ and DQ (Rs = 0.44), while DQ showed stronger concordance with MQ (Rs = 0.63). Single-cell validation against expert ground truth demonstrated high performance of DQ in tumor cell detection, with sensitivity of 0.98.5, specificity of 0.99, and accuracy of 0.99, based on 27,958 annotated cells across 60 regions of interest comparable to microscopic high-power fields. Analysis of pre-analytical and analytical factors identified specimen type and cautery/crush artifacts as the main pre-analytical contributors to DQ-VQ discrepancies, while overall variations in specimen cellularity was the dominant analytical factor. CONCLUSION:In summary, this study provides the first comprehensive real-world evaluation of AI-based TCC quantification in routine molecular pathology workflow, highlighting its robustness, accuracy, and the critical role of pre-analytical standardization, as well as pathologists` oversight for successful clinical implementation.
PURPOSE:This study aimed to provide a detailed clinicopathological, molecular genetics, and immune landscape analysis of a large cohort of primary thoracic NUT carcinomas to identify diagnostic challenges, uncover potential therapeutic targets, and define the spatial heterogeneity of the tumor immune microenvironment (TIME). MATERIALS AND METHODS:Fifty-three thoracic NUT carcinoma cases were analyzed using integrated immunohistochemistry, fluorescence in situ hybridization, targeted RNA sequencing, and multiplex immunofluorescence. RESULTS:Histologically, the majority of tumors exhibited pleomorphic presentations. Notably, we observed diverse, atypical morphological patterns, including squamous like, basaloid, neuroendocrine tumor like, and undifferentiated, pseudoglandular structure, and lymphoma-like presentations. Immunophenotypically, tumor cells consistently expressed keratins, p63 or p40 and NUT (100%), with focal expression of Syn and CgA in a subset. RNA sequencing identified NUTM1 fusion partners in 20 cases: BRD3::NUTM1, BRD4::NUTM1, NSD3::NUTM1, and notably, the first reported thoracic NUT carcinoma case with YAP1::NUTM1 fusion, which also exhibited SMARCA4 deficiency. Specifically, 2 cases exhibited pseudoglandular structure with myxoid stroma, 1 case displayed a predominantly lymphoma-like morphology, and 2 basaloid cases contained NSD3::NUTM1 fusions. Follow-up data were available for limited 33 patients (8 survivors and 25 deaths). Male sex was associated with improved survival in univariate analysis, whereas fusion type was not an independent prognostic factor. Furthermore, we characterized the TIME of thoracic NUT carcinoma (6 cases), demonstrating the presence of cytotoxic T cells, macrophages, and both mature and immature tertiary lymphoid structures. This heterogeneity in the TIME may contribute to the variable response to immunotherapy in NUT carcinoma. CONCLUSIONS:Primary thoracic NUT carcinoma exhibits significant pathological and immune microenvironmental heterogeneity, which was first described, as evidenced by the identification of a novel YAP1::NUTM1 fusion and diverse immune cell infiltrates. This heterogeneity warrants further investigation because it may inform prognostic stratification and the development of immune-targeted therapies.
PURPOSE:Biliary lesions, including ductular reactions and biliary obstruction, are involved in the pathophysiology of human hepatobiliary diseases. However, little is known about the three-dimensional (3D) biliary structures associated with these diseases. We examined pathological changes in 3D-structures in human chronic liver diseases using tissue-clearing methods. MATERIALS AND METHODS:Thirty autopsied liver tissues were obtained from 2 patients with advanced primary biliary cholangitis (PBC), 6 with cirrhosis (viral hepatitis, 3; alcoholic liver disease [ALD], 2; metabolic dysfunction associated steatotic liver disease [MASLD], 1), 4 with mild fibrosis (viral, 1; ALD, 1; MASLD, 2), and 18 with non-specific reactive changes (NSR). Liver tissues were fixed in formalin and cleared using the clear, unobstructed brain/body imaging cocktails methods or rapid and non-destructive tissue-clearing system. 3D-imaging of biliary structures were performed using immunostaining with Alexa 488-labelled anti-cytokeratin (CK)19 and light sheet fluorescence microscopy (LSFM). The number of CK19-positive biliary components per portal tract and the degrees of biliary networks in the portal tract, anastomoses, bridging formation, meandering, and caliber variation in 3D-images were evaluated semi-quantitatively. RESULTS:3D-biliary structures were clearly visualized by tissue-clearing methods and LSFM. The number of biliary components was significantly higher with cirrhosis than with NSR (p<0.05). The degrees of anastomoses and bridging formation were significantly increased in cirrhosis, compared to other groups (p<0.01). Degrees of biliary networks and meandering were significantly higher with cirrhosis than with mild fibrosis and NSR (p<0.01). The number of biliary components and other factors were not elevated in the two cases of advanced PBC examined. The number of biliary components and degrees of 3D-biliary features correlated with the results of various liver function tests (p<0.01). CONCLUSIONS:3D-imaging using the tissue-clearing methods may be a promising strategy to examine pathological changes in human intrahepatic biliary structures in patients with various human hepatobiliary diseases.
PURPOSE:Gastrointestinal stromal tumors (GIST) are molecularly heterogeneous neoplasms defined by mutually exclusive driver alterations (KIT, PDGFRA, SDH, BRAF, RAS and NF1). However, driver mutations alone do not fully explain their biological and clinical variability. Chromosomal imbalances and loss of heterozygosity (LOH) may represent an additional layer of tumor characterization. We developed a single-nucleotide polymorphism (SNP)-based next-generation sequencing (NGS) panel enabling genome-wide LOH assessment from formalin-fixed paraffin-embedded tissue. MATERIALS AND METHODS:Forty-nine GIST cases molecularly classified by targeted NGS (KIT n=19, PDGFRA n=9, SDH-deficient n=8, NF1 n=7, quadruple wild type [qWT] n=6) were analyzed. LOH was inferred from variant allele frequency patterns across 1,826 genome-wide SNPs. RESULTS:Chromosome 14 was the most commonly affected (63%), followed by chromosomes 22 (45%), 15 (41%), 21 (27%), and 13 (20%). Loss of chromosome arm 1p occurred in 43% of tumors. Distinct subgroup-specific patterns emerged: KIT-mutant GIST exhibited the highest degree of genomic instability, whereas both SDH-deficient tumors and PDGFRA-mutant GIST displayed minimal chromosomal instability. NF1-mutant tumors showed recurrent single-arm chromosome 17 LOH. qWT GISTs were heterogeneous, including one case with extensive chromosomal instability. CONCLUSIONS:Genome-wide SNP-based LOH profiling reveals distinct, subgroup-specific patterns of chromosomal imbalance in GIST and may serve as a feasible complementary approach to driver mutation analysis for refined molecular characterization and potential future clinical utility.
PURPOSE:With trastuzumab deruxtecan demonstrating clinical benefit in human epidermal growth factor receptor 2 (HER2)-low and -ultralow breast cancer, precise discrimination at the lowest end of the HER2 immunohistochemistry (IHC) spectrum has become essential. In this 1-year retrospective study, we evaluated whether a commercially available, in vitro diagnostic-approved artificial intelligence (AI) system could improve the consistency of HER2 IHC scoring tested on all breast cancers from a single center. MATERIALS AND METHODS:In total, 853 HER2 IHC whole-slide images from 581 patients were analyzed and independently scored according to the American Society of Clinical Oncology/College of American Pathologists guidelines by 3 expert pathologists and 1 nonexpert reader, both without and with AI assistance. The AI provided categorical HER2 scores (0-3+) and quantitative cell-level staining metrics. Interobserver agreement was assessed using overall percent agreement (OPA), Conger's kappa (κ), and pairwise Cohen's κ. Logistic regression models were used to investigate factors associated with discrepancies between pathologists and the AI system. RESULTS:Before AI assistance, overall multirater agreement among human readers was substantial (OPA, 79.8%; 95% CI, 77.9-81.7; Conger's κ = 0.72, 95% CI, 0.69-0.74) and increased to near-perfect levels following AI assistance (OPA, 88.6%; 95% CI, 86.9-90.1; κ = 0.84; 95% CI, 0.82-0.86). Most discordances clustered around the 10% American Society of Clinical Oncology/College of American Pathologists cutoff and were driven mainly by undercalling of 2+ cases. Discordance was also associated with lower tumor cell counts, biopsy samples, and heterogeneous staining patterns. Among AI-classified HER2-0 tumors, true HER2-null cases without any detectable positive staining were rare, supporting a biological continuum of HER2 expression. CONCLUSIONS:AI-assisted HER2 IHC scoring significantly improves interobserver consistency and provides quantitative support that helps address key limitations of conventional categorical HER2 assessment.
PURPOSE:The transcription factor Ets-1 is widely implicated in hepatic carcinogenesis, yet its precise role in balancing fibrogenesis and regeneration remains poorly defined. This study elucidated the clinical relevance, upstream regulatory mechanisms, and dual cytoprotective/antifibrotic functions of Ets-1 during chronic liver injury and repair. MATERIALS AND METHODS:Ets-1 networks were evaluated using human cirrhotic tissues adjacent to hepatocellular carcinoma, a rat carbon tetrachloride fibrosis/recovery model, and Ets-1 knockout mice. Upstream regulation and epigenetic gating were characterized in vitro across human hepatocellular carcinoma lines via Western blotting, real-time reverse-transcription PCR, methylation-specific PCR, chromatin immunoprecipitation, and enzyme-linked immunosorbent assay. RESULTS:Hepatic Ets-1 expression was profoundly depleted in human and rodent fibrosis, inversely correlating with disease severity, but rebounded robustly during regeneration, colocalizing with hepatocellular heme oxygenase-1 (HO-1). Ets-1 knockout mice exhibited exacerbated fibrosis, blunted HO-1 induction, and heightened injury markers under toxic stress. Mechanistically, in vitro analyses identified Ets-1 as a downstream effector of the proregenerative epidermal growth factor/extracellular signal-regulated kinase axis. Crucially, Ets-1 responsiveness to mitogens was restricted by promoter DNA hypermethylation, acting as an epigenetic gatekeeper in specific cell lines. Functionally, Ets-1 overexpression suppressed profibrogenic transforming growth factor beta 1 secretion and mitigated oxidative cytotoxicity via redox-sensitive recruitment to the HO-1 promoter. This significantly reduced macromolecular damage, as evidenced by attenuated 4-hydroxynonenal and 8-hydroxy-2'-deoxyguanosine adduct formation. CONCLUSIONS:Ets-1 acts as a pivotal molecular switch that integrates growth factor signaling with antioxidant defenses to facilitate hepatic recovery. Restoring Ets-1-mediated cytoprotection offers a potential therapeutic strategy for chronic liver disease, provided that tumorigenic risks are carefully managed via targeted hepatocyte-specific delivery systems.
Lymph node (LN) metastasis is a critical indicator of poor prognosis in breast cancer (BC). BC-derived exosomes influence intercellular communication within the tumor microenvironment, driving metastatic progression. However, the precise mechanisms by which exosomes facilitate LN metastasis in BC remain unclear. We performed small RNA sequencing on serum exosomes to identify Piwi-interacting RNAs (piRNAs) associated with BC LN metastasis. Functional investigations of exosomal piR-hsa-28212 included in vitro assays for migration and tube formation of human lymphatic endothelial cells (HLECs), alongside an in vivo footpad-popliteal LN metastasis model. Specific interactions between piR-hsa-28212 and TBX1 (T-box transcription factor 1), as well as TBX1 and VEGF receptor 3 (VEGFR3), were validated through luciferase reporter assays. The stability of TBX1 mRNA was analyzed by actinomycin D assay. RNA pulldown, RNA immunoprecipitation (RIP), and RNA fluorescence in situ hybridization (FISH) assays were conducted to explore the interaction between piR-hsa-28212 and METTL3. PiR-hsa-28212 was significantly upregulated in serum exosomes of BC patients with LN metastasis. Exosomal piR-hsa-28212 enhanced HLECs migration and tube formation in vitro and promoted lymphangiogenesis and LN metastasis in vivo. Mechanistically, exosomal piR-hsa-28212 transferred from BC cells to HLECs stabilized TBX1 mRNA, thereby upregulating VEGFR3 expression. In BC cells, piR-hsa-28212 directly bound to and stabilized METTL3 protein, modulating N6-methyladenosine (m6A) methylation of vascular endothelial growth factor C (VEGFC) mRNA and consequently increasing VEGFC expression and secretion. Collectively, these findings reveal an exosome-mediated piRNA regulatory mechanism that synergistically amplifies VEGFC/VEGFR3 signaling to drive LN metastasis in BC, highlighting piR-hsa-28212 as a potential therapeutic target. Trial Registration: KYLL-2022-338.
This study aimed to recapitulate adipose-tumor interactions within the colorectal cancer (CRC) tumor microenvironment (TME) and to elucidate the role of adipose-derived stem cells (ADSCs) in regulating CRC progression through cytokine-mediated signaling. Human ADSCs were isolated from adipose tissue and directly cocultured with CRC cells using an oxygen-permeable, PDMS-based 3D coculture chip, followed by cytokine profiling of conditioned media, immunofluorescence analysis of spheroids, FACS-based cell separation, and molecular analyzes including western blotting, qPCR, and immunoprecipitation to interrogate HIF-1-related mechanisms. The results revealed that cancer-associated ADSCs secrete CCL8, which markedly enhances CRC cell migration. Mechanistically, ADSC-derived CCL8 activated the ERK signaling pathway in CRC cells, leading to increased HIF-1α protein accumulation without significant changes in protein stability. This was accompanied by enhanced interaction between HIF-1α and the transcriptional cofactor p300. Consequently, HIF-1α transcriptional activity was increased, resulting in the upregulation of downstream epithelial-mesenchymal transition markers and promoting a pro-migratory and aggressive cancer phenotype. These effects were particularly pronounced in the coculture system, where intensified crosstalk between ADSCs and cancer cells amplifies oncogenic signaling within the TME. Collectively, these findings demonstrate that ADSC-derived CCL8 functions as a key mediator of adipose-tumor crosstalk in CRC progression by driving HIF-1α-dependent signaling pathways. Furthermore, the PDMS-based 3D coculture platform employed in this study provides a robust and physiologically relevant experimental system for dissecting complex cell-cell interactions and cytokine-driven mechanisms within the TME of CRC.
Nedaplatin is a second-generation platinum compound with reduced nephrotoxicity compared with cisplatin. Given its favorable tolerability in advanced non-small cell lung cancer (NSCLC), we prospectively evaluated the tolerability and efficacy of an adjuvant nedaplatin plus docetaxel chemotherapy regimen in patients with completely resected NSCLC. Patients with stage IB-IIIA NSCLC who had undergone complete surgical resection received nedaplatin (80 mg/m2) and docetaxel (60 mg/m2) intravenously on day 1 once every 4 weeks for up to four cycles. The primary endpoint was treatment tolerability, defined as completion of the planned four cycles, and compared with historical data for the standard cisplatin plus vinorelbine regimen. The secondary endpoint was efficacy, determined by relapse-free survival (RFS) after surgery. Thirty-five patients were enrolled, with median age of 65 years (range, 36-77 years). Adenocarcinoma (57.1%) and stage II disease (60.0%) were most common. Dose reductions were needed in some patients, however, 74.3% of patients (n = 26) completed all four cycles, meeting the primary endpoint. Grade 3 or 4 neutropenia and anorexia occurred in 67.8% and 9.0% of patients, respectively. No grade 3 or 4 renal dysfunction was noted. Concerning efficacy, the median RFS time was 63 months (95% confidence interval [CI], 20-NE), and the 5-year overall survival rate was 85.7% (95% CI, 70.0-93.9). Long-term retrospective follow-up (median: 119 months) showed that the 10-year RFS rate was 52.9% (95% CI, 36.1-69.1). These results indicate that adjuvant chemotherapy with nedaplatin plus docetaxel is tolerable and effective in patients with completely resected NSCLC. Trial Registration: UMIN number: 000004634.
AIMS:Granulomatous lobular mastitis (GLM) is a rare, chronic, benign inflammatory disease of the breast with an unclear aetiology. This study aimed to characterise the microbial features of GLM using metagenomic next-generation sequencing (mNGS) and to provide potentially relevant microbial clues for clinical evaluation. METHODS:Twenty fresh lesion tissue samples were collected from 15 female patients with GLM, including one representative sample per patient and five additional deep tissue samples. Clinical data collection, mNGS, bioinformatics analysis and data interpretation were performed to characterise the microbial profiles of GLM lesions. RESULTS:In this study, all patients presented with palpable breast masses, breast pain and abscess formation. More than half showed increased white blood cell counts, neutrophil percentages, C reactive protein levels and erythrocyte sedimentation rates together with decreased lymphocyte percentages. Based on genus-level filtering, mNGS identified 16 bacterial genera, 14 fungal genera and 3 viral genera, revealing a complex but bacteria-dominated microbial profile. The most frequently detected bacterial genera were Corynebacterium, Cutibacterium, Acinetobacter, Staphylococcus and Hathewaya, with marked interpatient variation in relative abundance, while fungal profiles were relatively more concentrated. In five patients with both superficial and deep tissue samples, microbial profiles differed across sampling depths, particularly for bacterial composition. CONCLUSIONS:mNGS revealed a complex, bacteria-dominated microbial profile in GLM lesions and indicated that sampling depth may influence the detected microbial profiles. These findings may provide useful clues for clinical evaluation, but the pathogenic significance of these micro-organisms remains to be elucidated.
Analytical sensitivity is a fundamental property of all biomarker assays, yet its relationship to the diagnostic and clinical sensitivity of immunohistochemistry biomarkers has only recently begun to be defined. Nationwide Dutch comparison of HER2 immunohistochemistry performance reported a significant correlation between the lower limit of detection (LOD) and staining intensity in low-expressing cores, but no association between LOD and registry-derived HER2-low prevalence. This editorial explains why that null finding should not be interpreted as evidence that analytical sensitivity is unimportant for diagnostic or clinical sensitivity. The following 3 methodological constraints limit the ability of this design to reveal such an association: (1) restriction of LOD values to a narrow high-sensitivity band, (2) ecological correlation of laboratory-level LOD with aggregated case-level categorical scores, and (3) substantial misclassification inherent in the broad, only moderately reproducible HER2-low endpoint. Prior studies measuring LOD and dynamic range show that LOD materially affects low-level HER2 measurement and remains central to fit-for-purpose HER2-low testing.
PURPOSE:The purpose of this study was to determine the frequency and spectrum of FGFR3 alterations in urothelial carcinoma and to evaluate their association with histologic variants. MATERIALS AND METHODS:A total of 190 cases of urothelial carcinoma were analyzed for alterations in FGFR3. RESULTS:After consolidation at the patient level, 33 (17%) patients harbored FGFR3 alterations. S249C (n = 17; 52%) and Y373C (n = 8; 24%) were the predominant mutations, whereas FGFR3::TACC fusion events accounted for 4 (12%) cases. R248C (n = 3; 9%) and G370C (n = 1; 3%) were less frequent. Variant histology was identified in 25 of 33 (76%) tumors, with micropapillary differentiation being the most common pattern (n = 13; 39%), followed by squamous differentiation (n = 7; 21%). Both micropapillary tumors and tumors with squamous differentiation exclusively harbored S249C or Y373C mutations. CONCLUSIONS:The consistent association of these variant patterns with S249C or Y373C mutations suggests that hotspot FGFR3 alterations can persist across morphologic contexts in noninvasive and invasive disease. Recognition of these variant histologies may therefore support consideration of targeted FGFR testing.
Boron neutron capture therapy (BNCT) requires selective delivery of boron compounds to tumor cells to achieve therapeutic efficacy. In this study, a BNCT strategy centered on the sodium-dependent multivitamin transporter (SMVT)-targeted biotin-conjugated boron compound, BBCIP, was evaluated, with selective incorporation of p-boronophenylalanine (BPA) according to tumor transporter characteristics. To examine transporter-dependent effects, the CRL1666 metastatic brain tumor model in rats was compared with an F98 glioma model, which exhibits distinct transporter expression profiles. The F98 model is characterized by dominant L-type amino acid transporter 1 (LAT1) expression and low SMVT levels, whereas the CRL1666 model shows relatively high SMVT expression. BBCIP demonstrated stable and quantifiable intratumoral boron pharmacokinetics in both models. In BNCT experiments, BBCIP alone showed limited therapeutic efficacy in the LAT1-dominant F98 model (median survival, 29 vs. 23 days in controls), whereas survival was further improved with the addition of BPA. In contrast, in the SMVT-high CRL1666 model, BBCIP-based BNCT significantly prolonged survival (30 vs. 15.5 days in controls), with only modest additional benefit from BPA. These findings indicate that the therapeutic contribution of BBCIP-based BNCT depends on tumor transporter expression and that BPA may be advantageous in tumors with low SMVT expression. Rather than relying on a single boron carrier, these findings support a biology-driven strategy in which boron agents are selected and combined according to tumor-specific characteristics.
The limited clinical efficacy of immune checkpoint inhibitors (ICIs) remains a major challenge in the treatment of bladder cancer (BCa). Here, we report that a ketogenic diet (KD) and its principal circulating metabolite, β-hydroxybutyrate (β-HB), suppress bladder tumor growth and enhance responses to PD-L1 inhibitor. In a syngeneic MB49 model, KD reduced tumor growth compared with normal diet (ND). KD increased plasma β-HB and was associated with higher intratumoral PD-L1 expression. Exogenous β-HB supplementation under ND recapitulated the antitumor effect of KD, further enhancing anti-PD-L1 efficacy. Mechanistically, β-HB increased intracellular reactive oxygen species (ROS) and disrupted mitochondrial membrane potential in bladder cancer cells, leading to ATP depletion and enhanced apoptosis. Besides, β-HB also upregulated PD-L1 in vitro and in vivo through GPR109A-JAK2-STAT3 signaling axis. Immune profiling of treated tumors showed increased infiltration and effector function of CD8+ T cells and NK cells and decreased immunosuppressive populations, changes that were further amplified when combined with anti-PD-L1. Together, our results indicate that KD and β-HB exert both tumor intrinsic and immune modulatory effects that sensitize bladder tumors to PD-L1 inhibitor, supporting further evaluation of ketogenic interventions as adjuvants to immunotherapy in BCa.
Transdifferentiation from follicular lymphoma (FL) to histiocytic/dendritic cell sarcoma (HDS) is rare and requires molecular confirmation of shared clonal origin. Targetable mutations such as BRAF V600E may offer therapeutic opportunities in such aggressive neoplasms. We report an exceptional case of untreated localised FL transdifferentiated to an HDS after 18 years. Shared BCL2 rearrangement and mutation profile confirmed a clonal link, while the HDS acquired an additional BRAF V600E mutation. Treatment with BRAF/MEK inhibitors yielded a sustained 18-month clinical response. The disease later relapsed as high-grade B-cell lymphoma with MYC and BCL2 rearrangements (HGBCL-MYC/BCL2), still harbouring the BRAF mutation. Complete remission was achieved with Rituximab, Cyclophosphamide, Hydroxydaunorubicin, Oncovin and Prednisone, but the double-hit lymphoma relapsed 14 months later.This case illustrates sequential transformation from FL to BRAF-mutated HDS with excellent response to BRAF/MEK inhibition, followed by evolution into HGBCL-MYC/BCL2 responding transiently to immunochemotherapy, emphasising the value of repeated histological and molecular reassessment in FL evolution.
