Laboratory Investigation最新文献

{"title":"Tumor Necrosis Factor-α–Dependent Inflammation Upregulates High Mobility Group Box 1 To Induce Tumor Promotion and Anti–Programmed Cell Death Protein-1 Immunotherapy Resistance in Lung Adenocarcinoma","authors":"Lifei Kang ,&nbsp;Jingjing Cao ,&nbsp;Wenli Guo ,&nbsp;Xiaohui Cui ,&nbsp;Yangxuan Wei ,&nbsp;Jiayu Zhang ,&nbsp;Feiran Liu ,&nbsp;Chenyang Duan ,&nbsp;Qiang Lin ,&nbsp;Ping Lv ,&nbsp;Zhiyu Ni ,&nbsp;Jing Zuo ,&nbsp;Haitao Shen","doi":"10.1016/j.labinv.2024.102164","DOIUrl":"10.1016/j.labinv.2024.102164","url":null,"abstract":"<div><div>Tumor-associated chronic lung inflammation depends on tumor necrosis factor (TNF)-α to activate several cytokines as part of an inflammatory loop, which plays a critical role in tumor progression in lung adenocarcinoma. High mobility group box 1 (HMGB1) is a cytokine that mediates inflammation. Whether TNF-α–induced inflammation regulates HMGB1 to contribute to tumor progression and promotion in lung adenocarcinoma remains unclear. Thus, human samples and a urethane-induced inflammation-driven lung adenocarcinoma (IDLA) mouse model were used to explore the involvement of HMGB1 in tumorigenesis and tumor progression and efficacy of anti–programmed cell death protein (PD)-1 immunotherapy. High levels of HMGB1 were observed in human lung adenocarcinoma associated with poor overall survival in patients. HMGB1 upregulation was positively correlated with TNF-α–related inflammation and TIM-3⁺ infiltration. TNF-α upregulated intracellular and extracellular HMGB1 expression to contribute to tumor promotion in A549 cells in vitro. Using a urethane-induced IDLA mouse model, we found HMGB1 upregulation was associated with increased TIM-3⁺ T-cell infiltration. Blocking TNF-α–dependent inflammation downregulated HMGB1 expression and inhibited tumorigenesis in the IDLA model. Anti-PD-1 treatment alone did not inhibit tumor growth in the TNF-α–dependent IDLA, whereas anti-PD-1 combined with TNF-α blockade overcame anti-PD-1 immunotherapy resistance. Furthermore, anti-PD-1 combined with anti-HMGB1 also inhibited tumor growth in IDLA, suggesting that increased HMGB1 release by TNF-α contributes to the resistance of anti-PD-1 immunotherapy in IDLA. Thus, tumor-associated TNF-α–dependent inflammation upregulated intracellular and extracellular HMGB1 expression in an inflammatory loop, contributing to tumor promotion and anti-PD-1 immunotherapy resistance in lung adenocarcinoma.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 2","pages":"Article 102164"},"PeriodicalIF":5.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142503128","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":"Complement Component 3 Promotes Regeneration of Olfactory Receptor Neurons","authors":"Hiroki Kuwazoe ,&nbsp;Hideki Sakatani ,&nbsp;Masamitsu Kono ,&nbsp;Shizuya Saika ,&nbsp;Norimitsu Inoue ,&nbsp;Muneki Hotomi","doi":"10.1016/j.labinv.2024.102200","DOIUrl":"10.1016/j.labinv.2024.102200","url":null,"abstract":"<div><div>Olfactory receptor neurons (ORNs) in the olfactory epithelium are characterized by high regenerative capacity even after birth, but the molecular mechanisms involved in ORN regeneration remain unclear. Complement component 3 (C3) has been shown to promote tissue regeneration, so we hypothesized that C3 activates innate immunity and also promotes the regeneration of ORNs. In this study, we investigate the role of C3 in ORN regeneration. We used C3 knockout (KO) and wild-type C57BL/6J mice in this study to examine the olfactory regeneration process for 42 days after methimazole-induced olfactory disorder. To compare the regeneration process after ORN damage between C3 KO and wild-type mice, we conducted olfactory behavioral tests and immunohistologic analysis and examined growth factors and inflammatory cell induction. C3 KO mice showed delayed olfactory recovery with lower olfactory epithelial thickness. In C3 KO mice, ORN maturation was delayed in association with increased accumulation of immature ORNs. In the normal ORN regeneration process, undesirable immature ORNs are produced and eliminated by apoptosis. C3 deficiency reduced neutrophils induced during ORN regeneration, suggesting the involvement of C3 in ORN regeneration through neutrophil-dependent elimination of undesired ORNs. C3 is therefore suggested to have promoted ORN regeneration by preventing the accumulation of immature ORNs. In addition, C3 may assist ORN maturation by participating in ORN axon selection such as synaptic pruning. Our results indicate that C3, which is activated during pathogen infection, also promotes recovery from ORN damage. These findings may lead to new therapeutic strategies for olfactory disorder.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 2","pages":"Article 102200"},"PeriodicalIF":5.1,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142710430","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":"Virtual Tissue Microarrays for Validating Digital Biomarker Analysis in Colorectal Carcinomaf","authors":"Margarita Melnikova Jørgensen ,&nbsp;Stephen Jacques Hamilton-Dutoit ,&nbsp;Jesper Bertram Bramsen ,&nbsp;Claus Lindbjerg Andersen ,&nbsp;Ida Elisabeth Holm","doi":"10.1016/j.labinv.2025.104098","DOIUrl":"10.1016/j.labinv.2025.104098","url":null,"abstract":"<div><div>Tissue microarrays (TMAs) are used for high-throughput biomarker discovery and validation. Although TMAs have numerous advantages, they may not always be representative of the tissue heterogeneity present in whole tissue sections (WTS) leading to inadequate biomarker quantification. In this pilot study, we studied biomarker expression in 50 randomly selected colorectal cancers and 36 microsatellite unstable cases with or without <em>BRAF</em> variants. We used virtual TMAs to determine the minimum number of tissue cores needed to quantify biomarkers with the same precision as when using WTS. Paraffin sections were immunohistochemically stained for markers of T cells, B cells, cancer-associated fibroblasts, and macrophages. Digitized WTS were divided into tumor center (TC) and invasive margin regions. The minimum number of virtual TMA cores in each region was determined by Bland-Altman plots with 95% limits of agreement. Bland-Altman plots showed substantial disagreement between TMAs and WTS, being highest for 3 cores and decreasing with increasing core numbers. However, even when using 8 cores, the limits of agreement between TMA and WTS were wide, indicating a high degree of measuring uncertainty using TMAs. When using 3 or 4 cores, TMAs underestimated the expression of all the biomarkers in the TC; similarly, levels of macrophage markers in the TC, and levels of B cells in both the TC and the invasive margin remained considerably underestimated, even when using the maximum number of cores possible. However, 3 cores were sufficient to adequately classify biomarkers into categoric low and high expression groups. Microsatellite unstable tumors were characterized by high heterogeneity, which was further increased in the presence of <em>BRAF</em> variant(s). The virtual TMA technique is a useful method to establish the minimum number of cores to be included when constructing tumor TMAs for biomarker analysis. Our results emphasize the importance of TMA validation for a specific biomarker prior to conducting larger clinical studies.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104098"},"PeriodicalIF":5.1,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143080562","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":"Microsatellite-Stable Gastric Cancer Can be Classified into 2 Molecular Subtypes with Different Immunotherapy Response and Prognosis Based on Gene Sequencing and Computational Pathology","authors":"Zhiyi Ye ,&nbsp;Xiaoyang Wu ,&nbsp;Zheng Wei ,&nbsp;Qiuyan Sun ,&nbsp;Yanli Wang ,&nbsp;Tan Li ,&nbsp;Yuan Yuan ,&nbsp;Jingjing Jing","doi":"10.1016/j.labinv.2025.104101","DOIUrl":"10.1016/j.labinv.2025.104101","url":null,"abstract":"<div><div>Most patients with gastric cancer (GC) exhibit microsatellite stability, yet comprehensive subtyping for prognostic prediction and clinical treatment decisions for microsatellite-stable GC is lacking. In this work, RNA-sequencing gene expression data and clinical information of patients with microsatellite-stable GC were obtained from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) databases. We employed several machine learning methods to develop and validate a signature based on immune-related genes (IRGs) for subtyping patients with microsatellite-stable GC. Moreover, 2 deep learning models based on the Vision Transformer (ViT) architecture were developed to predict GC tumor tiles and identify microsatellite-stable GC subtypes from digital pathology slides. Microsatellite status was evaluated by immunohistochemistry, and prognostic data as well as hematoxylin and eosin whole-slide images were collected from 105 patients with microsatellite-stable GC to serve as an independent validation cohort. A signature comprising 5 IRGs was established and validated, stratifying patients with microsatellite-stable GC into high-risk (microsatellite-stable-HR) and low-risk (microsatellite-stable-LR) groups. This signature demonstrated consistent performance, with areas under the receiver operating characteristic curve (AUC) of 0.65, 0.70, and 0.70 at 1, 3, and 5 years in the TCGA cohort, and 0.70, 0.60, and 0.62 in the GEO cohort, respectively. The microsatellite-stable-HR subtype exhibited higher levels of tumor immune dysfunction and exclusion, suggesting a greater potential for immune escape compared with the microsatellite-stable-LR subtype. Moreover, the microsatellite-stable-HR/LR subtypes showed differential sensitivities to various therapeutic drugs. Leveraging morphologic differences, the tumor recognition segmentation model achieved an impressive AUC of 0.97, whereas the microsatellite-stable-HR/LR identification model effectively classified microsatellite-stable-HR/LR subtypes with an AUC of 0.94. Both models demonstrated promising results in classifying patients with microsatellite-stable GC in the external validation cohort, highlighting the strong ability to accurately differentiate between microsatellite-stable GC subtypes. The IRG-related microsatellite-stable-HR/LR subtypes had the potential to enhance outcome prediction accuracy and guide treatment strategies. This research may optimize precision treatment and improve the prognosis for patients with microsatellite-stable GC.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104101"},"PeriodicalIF":5.1,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143080561","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":"Automated Scoring to Assess RAD51-Mediated Homologous Recombination in Ovarian Patient-Derived Tumor Organoids","authors":"Lucie Thorel ,&nbsp;Nicolas Elie ,&nbsp;Pierre-Marie Morice ,&nbsp;Louis-Bastien Weiswald ,&nbsp;Romane Florent ,&nbsp;Marion Perréard ,&nbsp;Florence Giffard ,&nbsp;Agathe Ricou ,&nbsp;Raphaël Leman ,&nbsp;Guillaume Babin ,&nbsp;Jean-François Lebrun ,&nbsp;Sandrine Martin ,&nbsp;Mélanie Briand ,&nbsp;Bernard Lambert ,&nbsp;Florence Joly ,&nbsp;Cécile Blanc-Fournier ,&nbsp;Dominique Vaur ,&nbsp;Enora Dolivet ,&nbsp;Benoit Plancoulaine ,&nbsp;Laurent Poulain","doi":"10.1016/j.labinv.2025.104097","DOIUrl":"10.1016/j.labinv.2025.104097","url":null,"abstract":"<div><div>Poly(ADP-ribose) polymerase inhibitors (PARPi) have been shown to improve progression-free survival, particularly in homologous recombination-deficient ovarian cancers. Identifying patients eligible for PARPi is currently based on next-generation sequencing, but the persistence of genomic scars in tumors after restoration of homologous recombination (HR) or epigenetic changes can be a limitation. Functional assays could thus be used to improve this profiling and faithfully identify homologous recombination-deficient tumors. The repair capacity (RECAP) test assesses the formation of RAD51 foci in proliferating cells after irradiation and can be used on tumors as well as on patient-derived tumor organoids (PDTO). However, RAD51 foci scoring is often performed manually without standardization. The purpose of this translational study was to develop an automated tool for scoring RAD51-mediated HR based on whole slide imaging of ovarian PDTO. To that end, we quantified Cyclin A2 and RAD51 immunofluorescence on 9 PDTO models derived from 8 ovarian cancer patients, and next, we compared the RECAP test results to genome instability score and to the patient clinical response. We therefore developed a standardized and automatized quantitative histoimaging tool allowing a comparative RAD51 foci evaluation and thus to define the HR status in PDTO. Our RECAP-based classification was correlated to the genome instability score, offering a new opportunity for standardization of HR assessment in PDTO. This new automated tool to score HR status, which remains to be validated on a large cohort of patients, may thus be used as a complement to next-generation sequencing-based tests in order to improve the identification of the number of patients eligible for PARPi.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104097"},"PeriodicalIF":5.1,"publicationDate":"2025-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143039224","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":"HMGB1 Encapsulated in Podocyte-Derived Exosomes Plays a Central Role in Glomerular Endothelial Cell Injury in Lupus Nephritis by Regulating TRIM27 Expression","authors":"Jinxi Liu ,&nbsp;Tongyu Zhao ,&nbsp;Huixin Cui ,&nbsp;Yuexin Tian ,&nbsp;Xinyan Miao ,&nbsp;Lingling Xing ,&nbsp;Xiaorong Wang ,&nbsp;Jie Huang ,&nbsp;Qingjuan Liu ,&nbsp;Wei Zhang ,&nbsp;Ke Shi ,&nbsp;Yunhe Liu ,&nbsp;Baiyun Jia ,&nbsp;Lihua Kang ,&nbsp;Yu Tian ,&nbsp;Weicheng Yuan ,&nbsp;Shiwei He ,&nbsp;Xiaojuan Feng ,&nbsp;Shuxia Liu","doi":"10.1016/j.labinv.2025.104096","DOIUrl":"10.1016/j.labinv.2025.104096","url":null,"abstract":"<div><div>Exosomes play a role in cell communication by transporting content between cells. Here, we tested whether renal podocyte-derived exosomes affect the injury of glomerular endothelial cells in lupus nephritis (LN). We found that exosomes containing high levels of high mobility group protein B1 (HMGB1) were released from podocytes in patients with LN, BALB/c mice injected with pristane (which induces lupus-like disease in mice), and cultured human renal glomerular endothelial cells (HRGECs) treated with LN plasma. In vitro, GW4869 (an inhibitor of exosome biogenesis/release) or exosome removal alleviated the injury of HRGECs induced by LN plasma. Additionally, leptomycin B or knockdown of HMGB1 in podocyte-derived exosomes reduced endothelial cell injury and the expression of tripartite motif-containing protein 27 (TRIM27). Knockdown or overexpression of TRIM27 attenuated or promoted the damage of HRGECs treated with LN plasma. In vivo, knockdown of HMGB1 in podocytes ameliorated the injury of glomerular endothelial cells in a mouse model of LN. Furthermore, the injection of podocyte-derived exosomes into mice caused glomerular endothelial cell dysfunction. In conclusion, our study revealed that podocyte-derived exosomes may mediate the injury of glomerular endothelial cells seen in LN.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104096"},"PeriodicalIF":5.1,"publicationDate":"2025-01-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143029128","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":"Monospecies Bacteria-Induced Chronic Apical Periodontitis Triggers the Aortic Inflammatory Response Via Modulation of Systemic Inflammation and Lipid Metabolism","authors":"Huaxiang Lei ,&nbsp;Shuai Chen ,&nbsp;Xiaojing Huang ,&nbsp;Dianfu Ma ,&nbsp;Yufang Luo ,&nbsp;Suli Xiao ,&nbsp;Pingping Li ,&nbsp;Guowu Gan ,&nbsp;Zhiyu Cai","doi":"10.1016/j.labinv.2025.104095","DOIUrl":"10.1016/j.labinv.2025.104095","url":null,"abstract":"<div><div>Cardiovascular disease (CVD) is the leading cause of death worldwide and has been confirmed to be associated with a common oral bacterial infection—chronic apical periodontitis (CAP). However, the detailed mechanisms remain controversial. CAP can potentially alter systemic inflammation, lipid metabolism, and gut microbiota, all of which contribute to the progression of the aortic inflammatory response. This study aimed to explore the differential effects between <em>Enterococcus faecalis</em> and <em>Porphyromonas gingivalis</em>-CAP on the aortic inflammatory response, which focused on changes in systemic inflammation, lipid metabolism, and gut microbiota, to explore potential mechanisms linking oral disease to CVD. Our results showed <em>P. gingivalis</em>-CAP could activate more serious aortic inflammatory cytokine micro RNA expression (TNF-α, MCP-1, and ICAM-1) than <em>E. faecalis</em>-CAP by promoting higher serum inflammation (TNF-α, IL-6, IL-1α, and MCP-1) and lipid (low-density lipoprotein cholesterol and total cholesterol) levels. Simultaneously, there was no significant change in gut microbiota between them. Furthermore, all serum inflammatory cytokines showed substantial correlations with aortic inflammatory cytokine micro RNA expression, and certain serum lipid indicators showed significant correlations, but only 2 gut microorganisms (<em>Ruminococcaceae</em> and <em>Prevotellaceae</em>) showed significant correlations. The combined results suggest that CAP might activate the aortic inflammatory response in association with changes in the 3 potential mechanisms. However, the promotion of gut microbiota might be relatively weak. Using experimental CAP induced by specific bacteria, in which bacteria are sequestered in the medullary cavity, avoids the direct influence of blood or intestinal pathways and provides new perspectives for studying the mechanism of CVD associated with oral disease. Overall, these findings suggest that CAP may exacerbate systemic inflammation and serum lipid levels in patients with CVD, highlighting the importance of educating such patients on oral hygiene.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104095"},"PeriodicalIF":5.1,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007961","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":"Molecular and Immunohistochemical Classification of Extrapulmonary Small Cell Neuroendocrine Carcinomas: A Study of 181 Cases","authors":"Klára Pavlíčková ,&nbsp;Jan Hojný ,&nbsp;Petr Waldauf ,&nbsp;Marián Švajdler ,&nbsp;Pavel Dundr ,&nbsp;Pavel Fabian ,&nbsp;Eva Krkavcová ,&nbsp;Jiří Dvořák ,&nbsp;Romana Michálková ,&nbsp;Iva Staniczková Zambo ,&nbsp;Nikola Hájková ,&nbsp;Miroslava Flídrová ,&nbsp;Jan Laco ,&nbsp;Helena Hornychová ,&nbsp;Patricie Delongová ,&nbsp;Jozef Škarda ,&nbsp;Jan Hrudka ,&nbsp;Radoslav Matěj","doi":"10.1016/j.labinv.2025.104093","DOIUrl":"10.1016/j.labinv.2025.104093","url":null,"abstract":"<div><div>Extrapulmonary small cell neuroendocrine carcinoma (EP-SCNC) is a rare malignancy with a poor prognosis. Most patients with EP-SCNC have metastatic disease upon presentation, and their average overall survival (OS) is less than 12 months. Our study aimed to conduct a complex analysis of EP-SCNC. One hundred eighty-one EP-SCNC tissue samples were subjected to a complex analysis. One hundred fifty-five tumors were pure EP-SCNC, whereas 26 were combined tumors. Immunohistochemistry for ASCL1, NEUROD1, YAP1, POU2F3, Rb1, p53, cyclin D1, p16, PTEN, DLL3, PD-L1, CD56, synaptophysin, chromogranin A, and INSM1 was performed, and 128 samples were analyzed molecularly using next-generation sequencing, comprising DNA and RNA analyses. Detailed results on immunohistochemical and molecular analyses were provided for each primary origin of EP-SCNC separately. Median survival for the whole cohort of patients was 8.94 months. Patient age (≥70 years), tumor mutational burden &lt;15, and <em>TP53</em> and <em>BRCA2</em> mutations were negative prognostic factors. High expression of ASCL-1 was associated with shorter OS, whereas high expression of YAP1 was associated with longer OS. Patients with genitourinary tumors had significantly better OS than those with gastrointestinal tract EP-SCNC tumors. Rb1 expression loss was detected more often in genitourinary tract SCNCs. In contrast, p16 overexpression was found more often in genitourinary tract SCNCs. POU2F3 expression was detected more often in combined tumors, whereas NEUROD1 was detected more often in pure EP-SCNC. Regarding “druggable markers,” DLL3 was expressed in 66% of tumors and PD-L1 in 17.4%. Detailed analyses of different prognostic and predictive markers are needed to better understand EP-SCNC biology and create more personalized therapy to improve patient prognosis.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104093"},"PeriodicalIF":5.1,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007960","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":"Leveraging Deep Learning for Immune Cell Quantification and Prognostic Evaluation in Radiotherapy-Treated Oropharyngeal Squamous Cell Carcinomas","authors":"Fanny Beltzung ,&nbsp;Van-Linh Le ,&nbsp;Ioana Molnar ,&nbsp;Erwan Boutault ,&nbsp;Claude Darcha ,&nbsp;François Le Loarer ,&nbsp;Myriam Kossai ,&nbsp;Olivier Saut ,&nbsp;Julian Biau ,&nbsp;Frédérique Penault-Llorca ,&nbsp;Emmanuel Chautard","doi":"10.1016/j.labinv.2025.104094","DOIUrl":"10.1016/j.labinv.2025.104094","url":null,"abstract":"<div><div>The tumor microenvironment plays a critical role in cancer progression and therapeutic responsiveness, with the tumor immune microenvironment (TIME) being a key modulator. In head and neck squamous cell carcinomas (HNSCCs), immune cell infiltration significantly influences the response to radiotherapy (RT). A better understanding of the TIME in HNSCCs could help identify patients most likely to benefit from combining RT with immunotherapy. Standardized, cost-effective methods for studying TIME in HNSCCs are currently lacking. This study aims to leverage deep learning (DL) to quantify immune cell densities using immunohistochemistry in untreated oropharyngeal squamous cell carcinoma (OPSCC) biopsies of patients scheduled for curative RT and assess their prognostic value. We analyzed 84 pretreatment formalin-fixed paraffin-embedded tumor biopsies from OPSCC patients. Immunohistochemistry was performed for CD3, CD8, CD20, CD163, and FOXP3, and whole slide images were digitized for analysis using a U-Net-based DL model. Two quantification approaches were applied: a cell-counting method and an area-based method. These methods were applied to stained regions. The DL model achieved high accuracy in detecting stained cells across all biomarkers. Strong correlations were found between our DL pipeline, the HALO Image Analysis Platform, and the open-source QuPath software for estimating immune cell densities. Our DL pipeline provided an accurate and reproducible approach for quantifying immune cells in OPSCC. The area-based method demonstrated superior prognostic value for recurrence-free survival, when compared with the cell-counting method. Elevated densities of CD3, CD8, CD20, and FOXP3 were associated with improved recurrence-free survival, whereas CD163 showed no significant prognostic association. These results highlight the potential of DL in digital pathology for assessing TIME and predicting patient outcomes.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 4","pages":"Article 104094"},"PeriodicalIF":5.1,"publicationDate":"2025-01-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143007959","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":"Immune Biomarkers on Tissue Microarray Cores Support the Presence of Adjacent Tertiary Lymphoid Structures in Soft Tissue Sarcoma","authors":"Elahe Shenasa ,&nbsp;Shelby Thornton ,&nbsp;Dongxia Gao ,&nbsp;Felix K.F. Kommoss ,&nbsp;Torsten O. Nielsen","doi":"10.1016/j.labinv.2025.104091","DOIUrl":"10.1016/j.labinv.2025.104091","url":null,"abstract":"<div><div>Immunotherapy has emerged as a new treatment modality in some soft tissue sarcomas, particularly for tumors associated with tertiary lymphoid structures (TLSs). These structures are functional lymphoid aggregates, and their presence is indicative of an active anticancer immune response in the tumor microenvironment. The assessment of TLS as a predictive biomarker at scale on patient specimens remains challenging. Although tissue microarrays (TMAs) could facilitate this assessment, it is unclear whether small microarray cores can represent and identify associated TLS responses.</div><div>We sought to use multiplex immunohistochemistry to identify key components of TLS: T cells, B cells, and dendritic cells. The multiplex panels (CD3, CD20, CD208, and PNAd) were applied to 80 cases both on TMAs and on their cognate available full-faced sections from epithelioid sarcoma and dedifferentiated/well-differentiated liposarcoma case series. TMAs were digitally scored for the number of immune cells using the HALO image analysis platform, and cognate full-faced sections were visually evaluated for the presence of TLS. An independent validation set of soft tissue sarcomas (N = 49) was stained with the CD3, CD20, and CD208, and scored by QuPath.</div><div>A combined immune marker (defined as the presence of more than 24% CD3+ T cells, or 0.51% CD20+ B cells, or &gt;0.14% CD208+ mature dendritic cells on tissue microarray cores) is highly specific (100%) and moderately sensitive (61%) to predict the existence of TLS on full-faced sections. The combined immune marker showed a sensitivity of 25% and specificity of 91% on the validation set.</div><div>The combined immune marker assessed on tissue microarrays is highly specific in inferring the presence of TLS on cognate full-faced sections. Therefore, despite the small area sampled, tissue microarrays may be utilized to assess the clinical value of TLS on data sets where specificity is critical and large sample size can mitigate low-to-moderate sensitivity.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 3","pages":"Article 104091"},"PeriodicalIF":5.1,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142971594","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}