Laboratory Investigation最新文献

{"title":"Prevalence, Immune Checkpoint Expression, and Spatial Interplay of Immune Cells Are Linked to Favorable Tumor Phenotype in 4915 Human Carcinomas","authors":"Zhihao Huang ,&nbsp;Tim Mandelkow ,&nbsp;Jonas B. Raedler ,&nbsp;Elena Bady ,&nbsp;Jan H. Müller ,&nbsp;Ronald Simon ,&nbsp;Eik Vettorazzi ,&nbsp;Guido Sauter ,&nbsp;Julia Ebner ,&nbsp;Niclas C. Blessin","doi":"10.1016/j.labinv.2025.104248","DOIUrl":"10.1016/j.labinv.2025.104248","url":null,"abstract":"<div><div>Although there is rising evidence that immune cell subpopulations that are in direct contact with the tumor cells (intraepithelial) can predict response to immune checkpoint therapy and patients’ outcome, a comprehensive assessment of intraepithelial immune cells and their spatial interplay is lacking. To assess intraepithelial leukocyte densities, immune checkpoint expression, and spatial interactions in 43 carcinoma entities, 4915 tumor samples in a tissue microarray format were analyzed using a deep learning framework and BLEACH&amp;STAIN multiplex fluorescence immunohistochemistry. This approach enabled single-cell resolution quantification of 21 biomarkers through 7 sequential staining and imaging rounds. Immune and tumor cells were classified into 54 subpopulations. The mean intraepithelial immune cell density of CD8⁺ cytotoxic T cells, CD4⁺ T-helper cells, FOXP3⁺ regulatory T cells, CD20⁺ B cells, M1/M2 macrophages, and CD11c⁺ dendritic cells varied markedly between tumor entities and individual tumors. For instance, 88(±90) cells/mm² were found in tubular breast cancer, 661(±729) cells/mm² in colorectal cancer, and up to 2325(±2131) cells/mm² in squamous cell cancers from various origins. Unsupervised cluster analysis revealed a “cluster a” of 634 patients from almost all different tumor entities with an exceptionally high density of intraepithelial immune cells that was characterized by a unique interaction profile along with the highest immune checkpoint expression. Across all analyzed tumor entities, the intraepithelial highly inflamed cluster a was significantly linked to low pathologic tumor stage (<em>P</em> &lt; .001). The data from this study provide a comprehensive characterization of intraepithelial immune cells across 43 different human carcinomas and identified an inflamed pan-cancer phenotype characterized by strong interactions of intraepithelial CD8⁺ cytotoxic T cells, CD4⁺ T cells, dendritic cells, and M2 macrophages, along with highest levels of TIM3, PD-1, and CTLA-4 expression that is linked to a favorable tumor phenotype.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104248"},"PeriodicalIF":4.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145258496","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Artificial Intelligence–Powered Spatial Analysis of the Tumor Microenvironment in Pulmonary Lymphoepithelial Carcinoma","authors":"Haohua Teng ,&nbsp;Yueyuxiao Yang ,&nbsp;Chan Xiang ,&nbsp;Jikai Zhao ,&nbsp;Zhanxian Shang ,&nbsp;Qian Zhu ,&nbsp;Lianying Guo ,&nbsp;Qiushun He ,&nbsp;Meng Yang ,&nbsp;Yuchen Han","doi":"10.1016/j.labinv.2025.104252","DOIUrl":"10.1016/j.labinv.2025.104252","url":null,"abstract":"<div><div>Lymphoepithelial carcinoma (LEC) can occur in various organs, such as the lung, nasopharynx, and thymus. We investigated the spatial characteristics of the tumor immune microenvironment (TIME) among primary pulmonary LECs (pLECs), pulmonary metastatic nasopharyngeal carcinomas (pmNPCs), and thymic LECs (tLECs). In this retrospective study, a total of 160 surgically resected LEC cases, comprising 116 pLECs, 26 tLECs, and 18 pmNPCs, were included. The TIME features, based on hematoxylin and eosin (H&amp;E)-stained whole-slide images (WSIs) and multiplexed immunofluorescence staining images, were obtained and their association with patient prognosis was analyzed. We developed a semisupervized model for automated tumor segmentation based on H&amp;E WSIs. The performance of the model was robust, with a mean accuracy rate of 0.847 on the testing set. Subsequent TIME analysis revealed different spatial distribution patterns of lymphocytes on H&amp;E WSIs among pLECs, tLECs, and pmNPCs. Lymphocyte count and distribution were prognostically relevant in pLECs, with an increasing trend of lymphocytes from the peripheral normal lung area to the tumor core in patients with a good prognosis. Further TIME analysis based on multiplexed immunofluorescence images uncovered that spatial arrangement and spatial interaction pattern characteristics were dependent on specific tumor types and cell subtypes. Our semisupervized learning model offers an automated and reproducible method for tumor segmentation for the TIME of rare LECs. Our analysis revealed different TIME patterns that distinguish among pLEC, tLEC, and pmNPC and demonstrates that the spatial arrangement and positional interaction patterns of PDL1⁺ tumor cells and FOXP3⁺ regulatory T cells could stratify prognosis in patients with pLEC.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104252"},"PeriodicalIF":4.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145301614","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessing the Status of Cyclin E1 (CCNE1) From Gene to Protein Level in Ovarian and Endometrial Carcinomas: A Systematic Review","authors":"Alexis Trecourt ,&nbsp;Catherine Genestie ,&nbsp;Alexander Valent ,&nbsp;Mojgan Devouassoux-Shisheboran ,&nbsp;Etienne Rouleau ,&nbsp;Elisa Yaniz-Galende ,&nbsp;Audrey Leformal ,&nbsp;Valeria Naim ,&nbsp;Alexandra Leary","doi":"10.1016/j.labinv.2025.104249","DOIUrl":"10.1016/j.labinv.2025.104249","url":null,"abstract":"<div><div>Twenty percent and 45.4% of high-grade ovarian carcinomas (OC) and endometrial carcinomas (EC) exhibit <em>CCNE1</em> amplification (<em>CCNE1</em>-amp), respectively, which is related to poor prognosis, but could serve as predictive biomarker for response to innovative targeted therapies. However, there is no consensus regarding how to evaluate the CCNE1 status (at the DNA, RNA, and/or protein level). Therefore, we conducted a systematic review of CCNE1 status testing in tubo-ovarian neoplasms and EC, comparing their performance for clinical purposes and highlighting the test’s interpretation criteria (CRD420250651291). Among the 734 records initially found on PubMed and Google Scholar, 48 reports were finally included. Molecular analyses and immunohistochemistry (IHC) were reported on 9774 tubo-ovarian neoplasms and 750 EC, and 6966 tubo-ovarian neoplasms and 856 EC, respectively. The most frequently morphological used method to detect <em>CCNE1</em>-amp was fluorescent in situ hybridization (13/16 studies, 81.3%), with quite consensual criteria to defined amplification (ie, <em>CCNE1</em>/chromosome 19 ratio ≥2, and/or &gt;8/≥8 copies of <em>CCNE1</em> per nucleus, and/or ≥4 <em>CCNE1</em> copies in ≥40% of cells). The proportion of tubo-ovarian neoplasms with CCNE1 immunohistochemical overexpression varied from 13.5% to 96%, and 14.6% to 86.1% in EC. The sensitivity and specificity of CCNE1 IHC to detect/exclude <em>CCNE1</em>-amp varied from 54.5% to 100% and 59.3% to 90.1%, respectively. Given the reported data, CCNE1 overexpression should be considered either when an H-score is ≥100 or when the staining is &gt;60% with &gt;5% of cells strongly stained. Both <em>CCNE1</em>-amp and CCNE1 overexpressions were associated with poor prognosis and with response to Wee1 and CDK2 inhibitors in high-grade serous OC (overall response rate up to 53%, objective response rate of 32%-40%). In contrast, <em>CCNE1</em> messenger RNA overexpression had no prognostic value. Thus, both <em>CCNE1</em>-amp detection by fluorescent in situ hybridization and CCNE1 protein levels quantification using IHC represent today the most validated tools to determine the CCNE1 status in OC/EC.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104249"},"PeriodicalIF":4.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145280579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MMP13-Expressing and COL11A1-Expressing Cancer-Associated Fibroblasts: Key Drivers of Esophageal Squamous Cell Carcinoma Progression and Prognostic Indicators","authors":"Shu Kato ,&nbsp;Yuki Kato ,&nbsp;Makoto Kodama ,&nbsp;Kouhei Yamamoto ,&nbsp;Asuka Furukawa ,&nbsp;Yoshihiro Nagase ,&nbsp;Rinka Miyashiro ,&nbsp;Minako Takagi ,&nbsp;Masayoshi Sakano ,&nbsp;Hisashi Fujiwara ,&nbsp;Kenro Kawada ,&nbsp;Yusuke Kinugasa ,&nbsp;Kenichi Ohashi","doi":"10.1016/j.labinv.2025.104247","DOIUrl":"10.1016/j.labinv.2025.104247","url":null,"abstract":"<div><div>The tumor microenvironment comprises various cell types, and cancer-associated fibroblasts (CAFs) are crucial contributors to cancer progression and metastasis. CAFs also play an important role in esophageal squamous cell carcinoma and have been extensively studied in this context. However, the association between CAFs and progression across pathological stages has not yet been reported. To identify these specific CAFs, we used a case-oriented approach for single-cell RNA sequencing. Consequently, we identified 3 CAF clusters classified as myofibroblastic CAFs (myCAFs), which increased in number as the cancer progressed. Pathway analysis revealed that the 3 CAF clusters had distinct properties. These CAFs were named <em>MMP13</em>⁺, <em>COL11A1</em>⁺, and <em>SFRP4</em>⁺ myCAFs based on their characteristic gene expression. We also investigated the distribution of various immune cells within the tumor microenvironment associated with the 3 different CAF clusters. The results revealed the presence of different types of immune cells, including M2 macrophages, regulatory T cells, and interferon gamma⁺ programmed death-1⁺ T cells and interferon gamma⁺ programmed death-1⁻ T cells. Next, we evaluated the presence of these 3 CAF subtypes in surgically resected specimens from patients with advanced esophageal squamous cell carcinoma using RNA in situ hybridization. Analysis of the association between these 3 CAF subtypes and prognosis showed that 2 subtypes (<em>MMP13</em>⁺ and <em>COL11A1</em>⁺ myCAFs) were associated with poor prognosis. <em>MMP13</em>⁺ myCAFs were associated with poorly differentiated infiltration patterns, whereas <em>COL11A1</em>⁺ myCAFs were associated with lymph node metastasis. These results suggest that future treatments targeting these CAFs and patient stratification based on these CAFs are warranted.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104247"},"PeriodicalIF":4.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145176288","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Combined Multicolor Immunofluorescence Staining and Spatial In Situ messenger RNA Expression Analysis Identifies Potential Fibrosis Drivers in Acute Lymphoblastic Leukemia","authors":"Sandro Bräunig ,&nbsp;Carl Dencker ,&nbsp;Dang Nghiem Vo ,&nbsp;Rong Fan ,&nbsp;Alba Lillo Sierras ,&nbsp;Jens Enoksson ,&nbsp;Anne Hultquist ,&nbsp;Hongzhe Li ,&nbsp;Stefan Scheding","doi":"10.1016/j.labinv.2025.104241","DOIUrl":"10.1016/j.labinv.2025.104241","url":null,"abstract":"<div><div>Acute lymphoblastic leukemia (ALL) is the most prevalent childhood cancer. Bone marrow (BM) fibrosis in ALL has been associated with adverse outcomes; however, little is known about the mechanisms that cause fibrosis in ALL. Therefore, we established a novel and advanced analysis method by combining multicolor immunofluorescence (IF) staining with in situ RNA expression analysis (RNAscope) to investigate the spatial expression of putative fibrotic drivers in ALL BMs. We analyzed standard BM biopsies from pediatric patients with ALL. Sequential 5-color IF staining with CD45, CD271, CD31, CD34, and DAPI was used to identify different BM cell types. Combined RNAscope and IF staining was established for spatial messenger RNA expression analysis of transforming growth factor beta 1 (<em>TGFB1</em>) and platelet-derived growth factor alpha 1 (<em>PDGFA1</em>), which are known to play major roles in primary myelofibrosis (PMF). PMF and normal BM samples served as controls. As expected, ALL BMs showed high cellularities and prominent populations of blast cells. CD271⁺ mesenchymal stromal cell density was increased in ALL and was associated with fibrosis in a similar manner as observed for PMF. <em>TGFB1</em> and <em>PDGFA1</em> expression was considerably increased in ALL megakaryocytes (MKs) compared with patients with PMF and normal controls. Furthermore, MK <em>TGFB1</em> and <em>PDGFA1</em> expression intensities in fibrotic ALL correlated with fibrosis grade. <em>TGFB1</em> and <em>PDGFA1</em> were also expressed in leukemic blasts, however, at lower intensities compared with ALL MKs. Taken together, advanced in situ RNA and IF staining not only revealed increased expression of <em>TGFB1</em> and <em>PDGFA1</em> in fibrotic pediatric ALL but also identified ALL blasts and MKs as their cellular origin at the single-cell level. These novel data strongly suggest a role of these cytokines as potential fibrosis drivers in ALL. More broadly, our findings demonstrate that combined RNA and surface marker analysis is a powerful tool to provide new and valuable insights into BM pathophysiology.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104241"},"PeriodicalIF":4.2,"publicationDate":"2025-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145103029","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Senescent Epithelial Cells Serve as Invasive Growth Drivers in Ameloblastoma","authors":"Hao Lin ,&nbsp;Jia-Jie Liang ,&nbsp;Chen-Xi Zhang ,&nbsp;Qi-Wen Man ,&nbsp;Rui-Fang Li ,&nbsp;Lin-Zhou Zhang ,&nbsp;Bing Liu","doi":"10.1016/j.labinv.2025.104227","DOIUrl":"10.1016/j.labinv.2025.104227","url":null,"abstract":"<div><div>Cellular senescence and its associated microenvironment play a pivotal role in tumor initiation and progression. Although ameloblastoma (AM) is classified as a benign tumor, it is characterized by local invasiveness and a high recurrence rate. In this study, we identified a distinct population of senescent epithelial cells using senescence-associated β-galactosidase staining and explored the role of this subpopulation in AM progression. The CellChat tool was utilized to map intercellular communication networks, revealing that this senescent cell cluster promotes stemness in neighboring cells and drives angiogenesis and osteoclastogenesis, which was subsequently confirmed by a series of <em>in vitro</em> experiments. Moreover, conditioned medium from these senescent cells significantly enhanced tumor growth in patient-derived organoids. Clinical data further demonstrated that elevated levels of cellular senescence were strongly associated with greater tumor invasiveness and poorer prognosis in AM patients. In conclusion, our findings suggest that targeting this specific subset of senescent epithelial cells may offer a novel therapeutic approach for AM management, potentially reducing tumor aggressiveness and recurrence.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104227"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804359","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tenascin-C Expression in Relation to Tumor-Stroma Interaction in Ameloblastoma","authors":"Satoko Sumi ,&nbsp;Shohei Yoshimoto ,&nbsp;Kanako Suyama ,&nbsp;Masahide Taguchi ,&nbsp;Hiromitsu Morita ,&nbsp;Akimitsu Hiraki ,&nbsp;Kyoko Oka","doi":"10.1016/j.labinv.2025.104237","DOIUrl":"10.1016/j.labinv.2025.104237","url":null,"abstract":"<div><div>Ameloblastoma (AM) is a benign epithelial odontogenic tumor that occurs in the jawbone. Although benign, AM can exhibit aggressive features, including locally invasive growth. In addition, local recurrence or distant metastasis may occur. Therefore, understanding the mechanisms underlying AM-cell migration is essential for improving clinical therapy. The role of the tumor microenvironment in disease progression has been extensively studied in various tumors. In the microenvironment, it has been reported that the extracellular matrix plays many roles. Tenascin-C (TN-C) is an extracellular matrix glycoprotein that is highly expressed during tissue development and remodeling. In this study, we investigated the involvement of TN-C in AM progression. Immunohistochemical analysis of AM specimens revealed high TN-C protein expression in the stroma, particularly at the invasive front. In contrast, RNA in situ hybridization demonstrated that <em>TNC</em> was localized within tumor cells, suggesting that the TN-C protein in the stroma is secreted by tumor cells rather than produced by stromal cells. In in vitro analyses, TN-C expression was significantly upregulated in cocultures of the AM cell line, AM-1, and primary human periodontal ligament fibroblasts, indicating that tumor-stroma interactions enhance tumor-derived TN-C expression. Functionally, TN-C stimulation promoted AM-cell migration, whereas <em>TNC</em> knockdown suppressed it. Spatial transcriptomics revealed elevated <em>TNC</em> expression in regions undergoing malignant transformation. Our results demonstrate that tumor-derived TN-C promotes AM progression. The expression of TN-C at the invasive front and in malignant regions suggests its potential as both a prognostic marker and a therapeutic target in tumor progression.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104237"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145006405","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fatty Pancreas Disease: An Integrated Study on Frozen Tissues Shows Distinct Compartments of Interlobular/Intralobular, Intra-Acinar, and Intra-Islet Fat Deposition","authors":"Claudio Luchini ,&nbsp;Carlotta Franzina ,&nbsp;Federico Caldart ,&nbsp;Nicolò De Pretis ,&nbsp;Manola Crestani ,&nbsp;Massimo Donadelli ,&nbsp;Paola Mattiolo ,&nbsp;Alessandra Fiore ,&nbsp;Federica Danzi ,&nbsp;Riccardo De Robertis ,&nbsp;Michele Bevere ,&nbsp;Roberto Baldan ,&nbsp;Laura Tommasi ,&nbsp;Nicolò Vianini ,&nbsp;Paolo Bernardi ,&nbsp;Mirco Galiè ,&nbsp;Antonio Pea ,&nbsp;Rachele Ciccocioppo ,&nbsp;Mirko D’Onofrio ,&nbsp;Roberto Salvia ,&nbsp;Luca Frulloni","doi":"10.1016/j.labinv.2025.104214","DOIUrl":"10.1016/j.labinv.2025.104214","url":null,"abstract":"<div><div>Obesity-related diseases and perturbations of fat metabolism represent some of the most common health challenges. In this complex scenario, recent evidence has suggested the emergence of a condition related to fat accumulation in the pancreas, which is generally referred to as fatty pancreas disease. This study aimed to clarify the different compartments of intrapancreatic fat deposition. The study cohort is represented by 100 patients who underwent pancreatic surgical resection. The pancreatic neck margin was analyzed with hematoxylin and eosin for evaluating tissue composition and with Oil Red O, a fat-specific histochemical staining, highlighting lipid droplets as red signals, for evaluating the presence of intracellular fat. Two cases were also analyzed with electron microscopy as cross-sectional validation. Regarding tissue composition, the most prevalent component was normal pancreatic parenchyma (mean value, 71.8%), followed by fibrosis (17.3%) and interlobular/intralobular fat (10.9%). Regarding intracellular fat deposition, Oil Red O–positive intracytoplasmic lipid droplets were present in most patients. The tissue areas with the highest levels of fat deposition were Langerhans’ islets, with neuroendocrine/insular cells showing more commonly a diffuse pattern of fat accumulation (&gt;75% of cells). Electron microscopy confirmed the presence of intracytoplasmic lipid vacuoles in neuroendocrine/insular cells. Our findings showed the presence of different compartments of intrapancreatic fat deposition, both in terms of tissue composition and intracellular compartmentalization. Understanding the mechanisms of fat deposition in the pancreas is crucial toward improving the general knowledge on fatty pancreas disease, also opening new perspectives for the study of lipid metabolism and the treatment of fat-related diseases.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104214"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144959249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comprehensive Analysis of Neurogenic Differentiation Factor 1 (NEUROD1), Achaete-Scute Homolog 1 (ASCL1), POU Class 2 Homeobox 3 (POU2F3), and Yes-Associated Protein 1 (YAP1) Expression Signatures Reveals Unique Large-Cell Neuroendocrine Carcinoma (LCNEC) Subgroups With Potential Therapeutic Implications","authors":"Frank W.J. Heijboer ,&nbsp;Jules L. Derks ,&nbsp;Dana A.M. Mustafa ,&nbsp;Nicole Rijnsburger ,&nbsp;Bregtje C.M. Hermans ,&nbsp;Lisa M. Hillen ,&nbsp;PALGA-Group ,&nbsp;Ernst-Jan M. Speel ,&nbsp;Anne-Marie C. Dingemans ,&nbsp;Jan H. von der Thüsen","doi":"10.1016/j.labinv.2025.104234","DOIUrl":"10.1016/j.labinv.2025.104234","url":null,"abstract":"<div><div>Large-cell neuroendocrine carcinoma (LCNEC) can be genomically subtyped into small-cell lung cancer (SCLC) and non-SCLC–like. Neurogenic differentiation 1 (NEUROD1), achaete-scute homolog 1 (ASCL1), POU class 2 homeobox 3 (POU2F3), and yes-associated protein 1 (YAP1) (NEUROD1, ASCL1, POU2F3, and YAP1 [NAPY]) subtypes have been reported for SCLC. We immunohistochemically evaluated NAPY in LCNEC alongside relevant protein expression data. Tissue microarrays from 133 stage I-III resected LCNEC were reviewed and immunostained for NAPY, protein retinoblastoma (pRb), delta-like ligand 3 (DLL3), cMYC, and thyroid transcription factor 1. An H-score of &gt;10 was considered positive (+), and &gt;50, dominant. Unsupervised clustering and spatial immune RNA profiling using GeoMx Digital Spatial Profiling (NanoString Technology) were performed. Clinical data were obtained from the Netherlands Cancer Registry. ASCL1 was dominant in 26% and NEUROD1 in 18% of LCNEC. pRb loss was observed in 75%, and DLL3+, cMYC+, and thyroid transcription factor 1+ in 66%, 26%, and 70%, respectively. Unsupervised clustering identified 5 expression-based subgroups: NEUROD1^high-ASCL1^high (10%), ASCL1^high (22%), POU2F3^high (5%), YAP1^high (11%), and NAPY^low (51%). Both ASCL1^high subgroups correlated with DLL3^high and high neuroendocrine (NE) marker expression. YAP1^high was enriched for pRb+. POU2F3^high and YAP1^high subgroups were NE marker low and DLL3^low. GeoMX Digital Spatial Profiling identified 4 upregulated genes involved in immune system and/or tumor development in the NEUROD1^high-ASCL1^high-POU2F3^high- group. In this comprehensive evaluation of NAPY markers in LCNEC, we observed 5 expression-based subgroups: NEUROD1^high-ASCL1^high, ASCL1^high, POU2F3^high, YAP1^high, and NAPY^low. The NE subgroups (NEUROD1^high-ASCL1^high and ASCL1^high) were recognized with DLL3^high expression. Compared with the proportion known in SCLC, more NAPY^low and YAP1^high and fewer POU2F3^high cases were identified. Application of NAPY in LCNEC provides a more modest discrimination of subgroups compared with SCLC. Further research on potential drug targets is warranted, ie, differences in DLL3 and YAP1 expression could guide personalized treatment strategies.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104234"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144992935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Prevalence of Atypical and Subclonal p53 Immunohistochemistry Expression in Mismatch Repair Deficient and/or POLE-Mutant Endometrial Carcinomas with TP53 Mutation.","authors":"Jing Wang, Yumeng Cai, Jun Wang, Jiuyuan Fang, Junyi Pang, Hui Zhang, Junliang Lu, Zijuan Zhang, Huanwen Wu, Zhiyong Liang","doi":"10.1016/j.labinv.2025.104216","DOIUrl":"10.1016/j.labinv.2025.104216","url":null,"abstract":"<p><p>p53 Immunohistochemistry (IHC) is a reliable surrogate for determining TP53 mutation status in endometrial carcinomas (ECs). However, the correlation of p53 IHC patterns and TP53 mutation characteristics in mismatch repair deficiency (MMRd) and/or POLE-mutant ECs was not comprehensively investigated. In this study, we identified 4 p53 expression patterns in 40 MMRd and/or POLE-mutant ECs with TP53 mutations. Thirteen cases (33%) displayed a wild-type pattern. Nine cases (23%) showed atypical pattern, characterized by the presence of eye-catching clustered cells with strong nuclear staining or weak-to-moderate cytoplasmic staining, which were patchily distributed with blurred boundaries. Fourteen cases (35%) demonstrated subclonal pattern with distinct regions of wild-type and mutation-type staining, of which 3 cases were originally misdiagnosed as \"mixed EC.\" Only 4 (10%) cases exhibited typical aberrant pattern. Tumors with wild-type and atypical patterns were predominantly associated with MMRd and POLE mutations, respectively. Among 52 TP53 mutations identified, 75% were missense and 25% were truncating, predominantly in DNA-binding domain. Gain-of-function missense mutations were more frequent in cases with subclonal patterns, whereas non-gain-of-function missense mutations predominated in wild-type or atypical patterns. Concurrent mutations were present in 25% of cases and were more common in aberrant or atypical patterns. Interestingly, 2 POLE wild-type cases with subclonal MMR expression showed p53 overexpression across the entire tumor, complicating molecular subtyping. These findings highlight the prevalence of atypical and subclonal p53 expression patterns in MMRd and/or POLE-mutant ECs with TP53 mutations, aiding in accurate IHC interpretation and thus more precise EC histological and molecular classification.</p>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":" ","pages":"104216"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}