Pub Date : 2025-11-01Epub Date: 2025-09-02DOI: 10.1016/j.labinv.2025.104236
Victoria Mahar, Zachary Colburn, Joshua Sakai
Challenging pathology case consultations require the shipment of irreplaceable patient materials to the consultants’ location for evaluation. In the military, consultants and generalists span geographically diverse locations. Shipped cases risk diagnostic delays, loss, and irreparable damage in transit over extensive distances. Using digital pathology for consultation eliminates these risks. Digital pathology implementation efforts in the Military Health System have been unsuccessful; however, triservice pathologists’ attitudes toward this innovation have never been investigated. Our explanatory mixed-methods study used a web-based needs assessment and interviews to understand pathologists’ facilitators and barriers to using digital pathology for consultation. We believe that understanding their perceptions is critical if further implementation efforts are to be successful. Analyses showed that pathologists were receptive to enterprise-wide implementation, especially if it improved turnaround time and allowed immediate subspecialist feedback. Future implementation efforts may benefit from comprehensive technical support combined with a consolidated digital pathology program office for implementation and sustainment guidance.
{"title":"Telepathology for Consultation in the Military Health System: An Evaluation of Pathologists’ Impressions of Facilitators and Barriers Prior to Implementation","authors":"Victoria Mahar, Zachary Colburn, Joshua Sakai","doi":"10.1016/j.labinv.2025.104236","DOIUrl":"10.1016/j.labinv.2025.104236","url":null,"abstract":"<div><div>Challenging pathology case consultations require the shipment of irreplaceable patient materials to the consultants’ location for evaluation. In the military, consultants and generalists span geographically diverse locations. Shipped cases risk diagnostic delays, loss, and irreparable damage in transit over extensive distances. Using digital pathology for consultation eliminates these risks. Digital pathology implementation efforts in the Military Health System have been unsuccessful; however, triservice pathologists’ attitudes toward this innovation have never been investigated. Our explanatory mixed-methods study used a web-based needs assessment and interviews to understand pathologists’ facilitators and barriers to using digital pathology for consultation. We believe that understanding their perceptions is critical if further implementation efforts are to be successful. Analyses showed that pathologists were receptive to enterprise-wide implementation, especially if it improved turnaround time and allowed immediate subspecialist feedback. Future implementation efforts may benefit from comprehensive technical support combined with a consolidated digital pathology program office for implementation and sustainment guidance.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104236"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145000987","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}
Pub Date : 2025-11-01Epub Date: 2025-08-25DOI: 10.1016/j.labinv.2025.104232
Lydia Kirsche , Lars Jansen , Annett Petzold , Petra Reinecke , Peter Behrens , Carol Geppert , Nikolaus Gaßler , Matthias Dürst
Human papillomavirus (HPV) DNA integration into the host genome is a frequent event in cervical carcinogenesis and may drive clonal expansion of the affected cells. Based on viral cellular junction (vcj) sequences, highly specific vcj-PCRs can be designed to detect viral integrants in DNA from cervical cell scrapes or tissue samples. In a recent study, such patient-specific vcj-PCR assays were employed for the detection of recurrent high-grade squamous intraepithelial lesions (HSIL) during postoperative surveillance. Although the specificity of vcj-PCR was 100%, only 50% of the recurrences were detected using this approach. The focus of the current study was to analyze the cause of this limited sensitivity. Using chemical microdissection and subsequent vcj-PCR analysis, we could demonstrate that the majority of lesions have a heterogeneous integrant pattern. Only 2 of 16 cones showed a homogeneous distribution of the respective integrants throughout the entire lesion. The other lesions displayed clonal outgrowths harboring the integrant in a background HPV16/18 DNA-positive HSIL tissue. In 4 cases, the respective integrant was undetectable in the lesion. These findings indicate that vcj-PCR has limited sensitivity for the detection of recurrent disease owing to intralesional heterogeneity. The observed heterogeneous integrant pattern may thus reflect the multifocal nature of most large HSIL. Alternatively, the possibility that HPV integration may be a late event in the carcinogenic process also needs to be considered.
{"title":"Heterogeneous Distribution of Human Papillomavirus (HPV) Integration Sites in Cervical Precancers Compromises the Diagnostic Accuracy of Integrant-Specific PCR","authors":"Lydia Kirsche , Lars Jansen , Annett Petzold , Petra Reinecke , Peter Behrens , Carol Geppert , Nikolaus Gaßler , Matthias Dürst","doi":"10.1016/j.labinv.2025.104232","DOIUrl":"10.1016/j.labinv.2025.104232","url":null,"abstract":"<div><div>Human papillomavirus (HPV) DNA integration into the host genome is a frequent event in cervical carcinogenesis and may drive clonal expansion of the affected cells. Based on viral cellular junction (vcj) sequences, highly specific vcj-PCRs can be designed to detect viral integrants in DNA from cervical cell scrapes or tissue samples. In a recent study, such patient-specific vcj-PCR assays were employed for the detection of recurrent high-grade squamous intraepithelial lesions (HSIL) during postoperative surveillance. Although the specificity of vcj-PCR was 100%, only 50% of the recurrences were detected using this approach. The focus of the current study was to analyze the cause of this limited sensitivity. Using chemical microdissection and subsequent vcj-PCR analysis, we could demonstrate that the majority of lesions have a heterogeneous integrant pattern. Only 2 of 16 cones showed a homogeneous distribution of the respective integrants throughout the entire lesion. The other lesions displayed clonal outgrowths harboring the integrant in a background HPV16/18 DNA-positive HSIL tissue. In 4 cases, the respective integrant was undetectable in the lesion. These findings indicate that vcj-PCR has limited sensitivity for the detection of recurrent disease owing to intralesional heterogeneity. The observed heterogeneous integrant pattern may thus reflect the multifocal nature of most large HSIL. Alternatively, the possibility that HPV integration may be a late event in the carcinogenic process also needs to be considered.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104232"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144959191","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}
Pub Date : 2025-11-01Epub Date: 2025-09-12DOI: 10.1016/j.labinv.2025.104239
Hetian Bai , Jinyong Li , Yeting Tu , Chongyun Bao
Cystic lesions are more prevalent in jawbones than in other bones. Odontogenic cysts are typically painless and asymptomatic and often reach significant sizes before detection. Unlike odontogenic cysts of inflammatory origin, understanding of the mechanisms of pathogenesis and progression of developmental odontogenic cysts remains incomplete. This study aimed to elucidate the cause and mechanisms of bone resorption in developmental odontogenic cysts. First, single-cell RNA sequencing was conducted for one of the most common developmental odontogenic cysts, dentigerous cysts (DCs), and the obtained data were compared with those of dental follicles of embedded teeth. Most DEGs in DCs and dental follicles were associated with immunoglobulin secretion, and an activated immunoglobulin-secreting plasma cell subtype was confirmed. Cell-to-cell interaction analysis revealed strong interactions between the activated plasma cells and leptin receptor–expressing (LepR+) fibroblasts via a “bone morphogenetic protein 6 and its receptor type 2" interaction. These LepR+ fibroblasts constituted the majority of fibroblasts in DCs. And these fibroblasts highly expressed genes were related to osteoclastogenesis, such as CSF1, IL6, and IL34. Mouse bone marrow–derived monocytes and bone marrow mesenchymal stem cells were treated with culture supernatants of the LepR+ fibroblasts. The treatment led to osteoclast formation and bone resorption, and inhibition of bone morphogenetic protein signaling suppressed the osteoclastogenesis effect. Thus, the LepR+ fibroblasts distinguished developmental odontogenic cysts from benign follicles; such cells interacted with activated plasma cell through the bone morphogenetic protein signaling pathway. And the LepR+ fibroblasts were crucial in osteoclast induction and bone resorption in DCs. This study confirmed a novel LepR+ fibroblast–induced bone erosion mechanism in developmental odontogenic cysts, which may inspire future pharmacological or surgical therapies.
{"title":"Leptin Receptor–Expressing (LepR+) Fibroblasts Activated by BMP Signaling Promote Bone Resorption in Developmental Odontogenic Cysts","authors":"Hetian Bai , Jinyong Li , Yeting Tu , Chongyun Bao","doi":"10.1016/j.labinv.2025.104239","DOIUrl":"10.1016/j.labinv.2025.104239","url":null,"abstract":"<div><div>Cystic lesions are more prevalent in jawbones than in other bones. Odontogenic cysts are typically painless and asymptomatic and often reach significant sizes before detection. Unlike odontogenic cysts of inflammatory origin, understanding of the mechanisms of pathogenesis and progression of developmental odontogenic cysts remains incomplete. This study aimed to elucidate the cause and mechanisms of bone resorption in developmental odontogenic cysts. First, single-cell RNA sequencing was conducted for one of the most common developmental odontogenic cysts, dentigerous cysts (DCs), and the obtained data were compared with those of dental follicles of embedded teeth. Most DEGs in DCs and dental follicles were associated with immunoglobulin secretion, and an activated immunoglobulin-secreting plasma cell subtype was confirmed. Cell-to-cell interaction analysis revealed strong interactions between the activated plasma cells and leptin receptor–expressing (LepR<sup>+</sup>) fibroblasts via a “bone morphogenetic protein 6 and its receptor type 2\" interaction. These LepR<sup>+</sup> fibroblasts constituted the majority of fibroblasts in DCs. And these fibroblasts highly expressed genes were related to osteoclastogenesis, such as <em>CSF1</em>, <em>IL6</em>, and <em>IL34</em>. Mouse bone marrow–derived monocytes and bone marrow mesenchymal stem cells were treated with culture supernatants of the LepR<sup>+</sup> fibroblasts. The treatment led to osteoclast formation and bone resorption, and inhibition of bone morphogenetic protein signaling suppressed the osteoclastogenesis effect. Thus, the LepR<sup>+</sup> fibroblasts distinguished developmental odontogenic cysts from benign follicles; such cells interacted with activated plasma cell through the bone morphogenetic protein signaling pathway. And the LepR<sup>+</sup> fibroblasts were crucial in osteoclast induction and bone resorption in DCs. This study confirmed a novel LepR<sup>+</sup> fibroblast–induced bone erosion mechanism in developmental odontogenic cysts, which may inspire future pharmacological or surgical therapies.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104239"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145065219","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}
The role of poly (adenosine diphosphate [ADP]-ribose) polymerase 1 binding protein (PARPBP) in hepatocellular carcinoma (HCC) prognosis remains unclear. This study investigated PARPBP expression in HCC clinical samples and evaluated its clinicopathological significance in relation to CD8-positive tumor-infiltrating lymphocytes (CD8+ TILs). PARPBP expression and CD8+ TIL density were assessed in surgical specimens from 96 patients with HCC. We performed immunohistochemical analysis to determine PARPBP protein levels, which were correlated with clinicopathological features and patient outcomes. Additionally, we performed experiments using a doxycycline-inducible short hairpin RNA system targeting PARPBP in Huh7 cells. Results indicated that moderately and poorly differentiated HCC had significantly higher PARPBP expression than well-differentiated tumors. Patients with high PARPBP expression exhibited shorter recurrence-free survival and overall survival than those with low expression (both P < .001). Multivariate analysis showed that high PARPBP expression (hazard ratio [HR], 2.806; P = .002), high CD8+ TIL density (HR, 0.148; P < .001), and α-fetoprotein ≥ 10 ng/mL (HR, 1.904; P = .047) were independent predictors of prognosis. Combined analysis revealed that patients with high PARPBP expression and low CD8+ TIL density had the worst recurrence-free survival and overall survival outcomes (both P < .001). In vitro experiments showed that the mRNA expression of PARPBP was higher in liver cancer cell lines than in normal liver cells and that PARPBP knockdown suppressed huh7 cell proliferation. In conclusion, elevated PARPBP expression was associated with poor differentiation and survival in patients with HCC. Furthermore, the combination of high PARPBP expression and low CD8+ TIL density improved prognostic accuracy.
{"title":"Prognostic Significance of Poly (ADP-Ribose) Polymerase 1 Binding Protein Expression and CD8+ Tumor-Infiltrating Lymphocytes in Hepatocellular Carcinoma","authors":"Masako Okada , Misako Sato-Matsubara , Masaru Enomoto , Truong Huu Hoang , Sawako Uchida-Kobayashi , Akihiro Tamori , Tsutomu Matsubara , Kenichi Kohashi , Takeaki Ishizawa , Norifumi Kawada","doi":"10.1016/j.labinv.2025.104226","DOIUrl":"10.1016/j.labinv.2025.104226","url":null,"abstract":"<div><div>The role of poly (adenosine diphosphate [ADP]-ribose) polymerase 1 binding protein (PARPBP) in hepatocellular carcinoma (HCC) prognosis remains unclear. This study investigated PARPBP expression in HCC clinical samples and evaluated its clinicopathological significance in relation to CD8-positive tumor-infiltrating lymphocytes (CD8<sup>+</sup> TILs). PARPBP expression and CD8<sup>+</sup> TIL density were assessed in surgical specimens from 96 patients with HCC. We performed immunohistochemical analysis to determine PARPBP protein levels, which were correlated with clinicopathological features and patient outcomes. Additionally, we performed experiments using a doxycycline-inducible short hairpin RNA system targeting PARPBP in Huh7 cells. Results indicated that moderately and poorly differentiated HCC had significantly higher PARPBP expression than well-differentiated tumors. Patients with high PARPBP expression exhibited shorter recurrence-free survival and overall survival than those with low expression (both <em>P</em> < .001). Multivariate analysis showed that high PARPBP expression (hazard ratio [HR], 2.806; <em>P</em> = .002), high CD8<sup>+</sup> TIL density (HR, 0.148; <em>P</em> < .001), and α-fetoprotein ≥ 10 ng/mL (HR, 1.904; <em>P</em> = .047) were independent predictors of prognosis. Combined analysis revealed that patients with high PARPBP expression and low CD8<sup>+</sup> TIL density had the worst recurrence-free survival and overall survival outcomes (both <em>P</em> < .001). In vitro experiments showed that the mRNA expression of <em>PARPBP</em> was higher in liver cancer cell lines than in normal liver cells and that <em>PARPBP</em> knockdown suppressed huh7 cell proliferation. In conclusion, elevated PARPBP expression was associated with poor differentiation and survival in patients with HCC. Furthermore, the combination of high PARPBP expression and low CD8<sup>+</sup> TIL density improved prognostic accuracy.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104226"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804358","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}
Pub Date : 2025-11-01Epub Date: 2025-09-02DOI: 10.1016/j.labinv.2025.104235
Ya-ting Qiu , Long-feng Ke , Wen-wen Zhang , Shu-yi Lu , Chen-yu Wu , Yun-li Xie , Yu Chen , Gang Chen , Yan-ping Chen
The BRAFV600E mutation test for melanoma patients has become the key to precision therapy. In this study, we compared the concordance of immunohistochemistry (IHC), quantitative real-time PCR (qPCR), and next-generation sequencing (NGS) in detecting the BRAFV600E mutation in a Chinese melanoma patient population. In addition, this study evaluated the BRAFV600E mutation heterogeneity between primary and metastatic melanoma sites, as well as within the same lesion, and investigated the association between BRAFV600E mutation status and tumor cell morphology. A total of 880 samples from 555 patients diagnosed with malignant melanoma were collected, and IHC for BRAFV600E was conducted. Of these, 385 were subjected to qPCR and 115 to NGS concurrently. Inter and intratumor heterogeneities of BRAFV600E mutations were compared. Hematoxylin and eosin (H&E) stain was performed, and the cell morphologies were reviewed. The IHC and qPCR results were discordant in 14 cases, yielding a concordance rate of 96.36%. IHC and NGS results showed a concordance rate of 97.39%. The sensitivity and specificity of BRAFV600E detection by IHC were 96.95% and 99.46%, with an overall concordance rate of 98.80%. One of 130 patients (0.77%) showed intertumor heterogeneity, and 3 of 880 samples (0.34%) showed intratumor heterogeneity. VE1 staining patterns significantly differed across cell morphologies (P < .01). Compared with qPCR and NGS, VE1 IHC offers high sensitivity, specificity, and consistency in detecting the BRAFV600E mutation in melanomas. The BRAFV600E mutation in melanoma exhibits low intertumor and intratumor heterogeneities and is significantly associated with tumor cell morphology; tumors with epithelioid cell morphology are most likely to harbor the BRAFV600E mutation.
{"title":"Association of the BRAFV600E Mutation With Morphology and Heterogeneity in Melanoma","authors":"Ya-ting Qiu , Long-feng Ke , Wen-wen Zhang , Shu-yi Lu , Chen-yu Wu , Yun-li Xie , Yu Chen , Gang Chen , Yan-ping Chen","doi":"10.1016/j.labinv.2025.104235","DOIUrl":"10.1016/j.labinv.2025.104235","url":null,"abstract":"<div><div>The <em>BRAFV600E</em> mutation test for melanoma patients has become the key to precision therapy. In this study, we compared the concordance of immunohistochemistry (IHC), quantitative real-time PCR (qPCR), and next-generation sequencing (NGS) in detecting the <em>BRAFV600E</em> mutation in a Chinese melanoma patient population. In addition, this study evaluated the <em>BRAFV600E</em> mutation heterogeneity between primary and metastatic melanoma sites, as well as within the same lesion, and investigated the association between <em>BRAFV600E</em> mutation status and tumor cell morphology. A total of 880 samples from 555 patients diagnosed with malignant melanoma were collected, and IHC for <em>BRAFV600E</em> was conducted. Of these, 385 were subjected to qPCR and 115 to NGS concurrently. Inter and intratumor heterogeneities of <em>BRAFV600E</em> mutations were compared. Hematoxylin and eosin (H&E) stain was performed, and the cell morphologies were reviewed. The IHC and qPCR results were discordant in 14 cases, yielding a concordance rate of 96.36%. IHC and NGS results showed a concordance rate of 97.39%. The sensitivity and specificity of BRAFV600E detection by IHC were 96.95% and 99.46%, with an overall concordance rate of 98.80%. One of 130 patients (0.77%) showed intertumor heterogeneity, and 3 of 880 samples (0.34%) showed intratumor heterogeneity. VE1 staining patterns significantly differed across cell morphologies (<em>P</em> < .01). Compared with qPCR and NGS, VE1 IHC offers high sensitivity, specificity, and consistency in detecting the <em>BRAFV600E</em> mutation in melanomas. The <em>BRAFV600E</em> mutation in melanoma exhibits low intertumor and intratumor heterogeneities and is significantly associated with tumor cell morphology; tumors with epithelioid cell morphology are most likely to harbor the <em>BRAFV600E</em> mutation.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104235"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145000891","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}
Pub Date : 2025-11-01Epub Date: 2025-07-17DOI: 10.1016/j.labinv.2025.104213
Anne Tuomisto, Päivi Sirniö, Hanna Elomaa, Henna Karjalainen, Ville K Äijälä, Meeri Kastinen, Vilja V Tapiainen, Maarit Ahtiainen, Olli Helminen, Erkki-Ville Wirta, Jukka Rintala, Sanna Meriläinen, Juha Saarnio, Tero Rautio, Toni T Seppälä, Jan Böhm, Jukka-Pekka Mecklin, Markus J Mäkinen, Juha P Väyrynen
High immune cell infiltration is generally associated with better survival in colorectal cancer (CRC). Recently, a prognostic score called CD8IE-FOXP3IS, which integrates the densities of tumor intraepithelial CD8+ and intrastromal FOXP3+ cells, was introduced using multiplex immunofluorescence. In this study, we developed a triple chromogenic immunohistochemistry assay to evaluate the CD8IE-FOXP3IS score and assessed its prognostic value in comparison with the CD3-CD8 T-cell density score (based on the principles of the Immunoscore) and conventional prognostic parameters. Multiplex immunohistochemistry combined with machine learning-assisted image analysis was used to quantify CD8IE and FOXP3IS densities in 2 independent cohorts comprising 1724 CRC patients. Multivariable Cox regression models were used to evaluate the prognostic value of the CD8IE-FOXP3IS score. We found that a low CD8IE-FOXP3IS score was associated with higher disease stage, more frequent lymphovascular invasion, and mismatch repair proficient status. In addition, a low CD8IE-FOXP3IS score was associated with higher CRC-specific mortality independent of the CD3-CD8 T-cell density score and other tumor and patient characteristics (cohort 1: hazard ratio [HR] for low vs high CD8IE-FOXP3IS score, 3.08; 95% CI, 1.54-6.15; Ptrend = 6.0E-4; cohort 2: HR, 4.30; 95% CI, 2.58-7.17; Ptrend = 3.2E-9). These findings indicate that triple chromogenic immunohistochemistry combined with digital pathology is an applicable method for quantifying tumor intraepithelial CD8+ and stromal FOXP3+ cell densities, allowing for the determination of the CD8IE-FOXP3IS score. The CD8IE-FOXP3IS score shows a strong prognostic value, which appears superior to overall CD3+ and CD8+ T-cell density measurement.
{"title":"Integrating Tumor Intraepithelial CD8<sup>+</sup> and Stromal FOXP3<sup>+</sup> T-Cell Densities as an Enhanced Immune Prognostic Index in Colorectal Cancer.","authors":"Anne Tuomisto, Päivi Sirniö, Hanna Elomaa, Henna Karjalainen, Ville K Äijälä, Meeri Kastinen, Vilja V Tapiainen, Maarit Ahtiainen, Olli Helminen, Erkki-Ville Wirta, Jukka Rintala, Sanna Meriläinen, Juha Saarnio, Tero Rautio, Toni T Seppälä, Jan Böhm, Jukka-Pekka Mecklin, Markus J Mäkinen, Juha P Väyrynen","doi":"10.1016/j.labinv.2025.104213","DOIUrl":"10.1016/j.labinv.2025.104213","url":null,"abstract":"<p><p>High immune cell infiltration is generally associated with better survival in colorectal cancer (CRC). Recently, a prognostic score called CD8<sup>IE</sup>-FOXP3<sup>IS</sup>, which integrates the densities of tumor intraepithelial CD8<sup>+</sup> and intrastromal FOXP3<sup>+</sup> cells, was introduced using multiplex immunofluorescence. In this study, we developed a triple chromogenic immunohistochemistry assay to evaluate the CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score and assessed its prognostic value in comparison with the CD3-CD8 T-cell density score (based on the principles of the Immunoscore) and conventional prognostic parameters. Multiplex immunohistochemistry combined with machine learning-assisted image analysis was used to quantify CD8<sup>IE</sup> and FOXP3<sup>IS</sup> densities in 2 independent cohorts comprising 1724 CRC patients. Multivariable Cox regression models were used to evaluate the prognostic value of the CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score. We found that a low CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score was associated with higher disease stage, more frequent lymphovascular invasion, and mismatch repair proficient status. In addition, a low CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score was associated with higher CRC-specific mortality independent of the CD3-CD8 T-cell density score and other tumor and patient characteristics (cohort 1: hazard ratio [HR] for low vs high CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score, 3.08; 95% CI, 1.54-6.15; P<sub>trend</sub> = 6.0E-4; cohort 2: HR, 4.30; 95% CI, 2.58-7.17; P<sub>trend</sub> = 3.2E-9). These findings indicate that triple chromogenic immunohistochemistry combined with digital pathology is an applicable method for quantifying tumor intraepithelial CD8<sup>+</sup> and stromal FOXP3<sup>+</sup> cell densities, allowing for the determination of the CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score. The CD8<sup>IE</sup>-FOXP3<sup>IS</sup> score shows a strong prognostic value, which appears superior to overall CD3<sup>+</sup> and CD8<sup>+</sup> T-cell density measurement.</p>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":" ","pages":"104213"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667952","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}
Pub Date : 2025-11-01Epub Date: 2025-10-02DOI: 10.1016/j.labinv.2025.104228
Lifei Kang , Jingjing Cao , Wenli Guo , Xiaohui Cui , Yangxuan Wei , Jiayu Zhang , Feiran Liu , Chenyang Duan , Qiang Lin , Ping Lvx , Zhiyu Ni , Jing Zuo , Haitao Shen
{"title":"Corrigendum to “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” [Laboratory Investigation 105 (2025) 102164]","authors":"Lifei Kang , Jingjing Cao , Wenli Guo , Xiaohui Cui , Yangxuan Wei , Jiayu Zhang , Feiran Liu , Chenyang Duan , Qiang Lin , Ping Lvx , Zhiyu Ni , Jing Zuo , Haitao Shen","doi":"10.1016/j.labinv.2025.104228","DOIUrl":"10.1016/j.labinv.2025.104228","url":null,"abstract":"","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104228"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145219929","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}
Pub Date : 2025-11-01Epub Date: 2025-07-29DOI: 10.1016/j.labinv.2025.104221
Takeshi Iwasaki, Mari Masunaga, Takumi Tomonaga, Yosuke Masumoto, Yuri Akiyama, Yoshinao Oda
N6-methyladenosine (m6A), a widespread RNA modification, plays a vital role in various biological processes, including carcinogenesis, tumor progression, and immune regulation. We conducted this study to investigate the relationship between m6A regulators, such as METTL3, METTL14, WTAP, FTO, ALKBH5, and YTHDF1-3, and their association with c-Myc and programmed death ligand 1 (PD-L1) expression in leiomyosarcoma (LMS). The expression of these epitranscriptome regulator genes was evaluated using the next-generation sequencing data of 53 patients with LMS obtained from an online public database. We next determined the relationship between m6A regulators and c-Myc and CD274 (PD-L1) mRNA expression in an LMS cell line. We also performed immunohistochemical staining of 69 LMS cases. Immunohistochemical staining showed that cases with higher expression of METTL3, METTL14, ALKBH5, FTO, and WTAP exhibited higher c-Myc expression, and cases with higher expression of ALKBH5, YTHDF2, and WTAP exhibited higher mitotic activity. Gene set enrichment analysis revealed that METTL3, METTL14, and FTO knockdown significantly suppressed c-Myc target gene expression. Knockdown of METTL3, ALKBH5, YTHDF1, WTAP, and FTO in LMS cell lines reduced cell proliferation. These results suggest the relationship between m6A modifications and c-Myc-driven oncogenesis. Moreover, knockdown of YTHDF2 inhibited interferon gamma–induced PD-L1 expression, suggesting its role in immune evasion through PD-L1 regulation. Multivariate Cox proportional hazards analysis revealed that lower YTHDF2 expression and higher WTAP expression are unfavorable prognostic factors. These findings provide potential therapeutic targets for LMS, particularly in combination with immune checkpoint inhibitors. Further investigation into the molecular mechanisms of m6A-mediated regulation of PD-L1 and c-Myc expression is required to develop more effective treatments for LMS.
{"title":"Comprehensive Analysis of N6-Methyladenosine (m6A) RNA Methylation Regulators in Soft Tissue Leiomyosarcoma","authors":"Takeshi Iwasaki, Mari Masunaga, Takumi Tomonaga, Yosuke Masumoto, Yuri Akiyama, Yoshinao Oda","doi":"10.1016/j.labinv.2025.104221","DOIUrl":"10.1016/j.labinv.2025.104221","url":null,"abstract":"<div><div>N6-methyladenosine (m6A), a widespread RNA modification, plays a vital role in various biological processes, including carcinogenesis, tumor progression, and immune regulation. We conducted this study to investigate the relationship between m6A regulators, such as METTL3, METTL14, WTAP, FTO, ALKBH5, and YTHDF1-3, and their association with c-Myc and programmed death ligand 1 (PD-L1) expression in leiomyosarcoma (LMS). The expression of these epitranscriptome regulator genes was evaluated using the next-generation sequencing data of 53 patients with LMS obtained from an online public database. We next determined the relationship between m6A regulators and <em>c-Myc</em> and <em>CD274 (PD-L1)</em> mRNA expression in an LMS cell line. We also performed immunohistochemical staining of 69 LMS cases. Immunohistochemical staining showed that cases with higher expression of METTL3, METTL14, ALKBH5, FTO, and WTAP exhibited higher c-Myc expression, and cases with higher expression of ALKBH5, YTHDF2, and WTAP exhibited higher mitotic activity. Gene set enrichment analysis revealed that <em>METTL3</em>, <em>METTL14</em>, and <em>FTO</em> knockdown significantly suppressed c-Myc target gene expression. Knockdown of <em>METTL3</em>, <em>ALKBH5</em>, <em>YTHDF1</em>, <em>WTAP</em>, and <em>FTO</em> in LMS cell lines reduced cell proliferation. These results suggest the relationship between m6A modifications and c-Myc-driven oncogenesis. Moreover, knockdown of <em>YTHDF2</em> inhibited interferon gamma–induced <em>PD-L1</em> expression, suggesting its role in immune evasion through PD-L1 regulation. Multivariate Cox proportional hazards analysis revealed that lower YTHDF2 expression and higher WTAP expression are unfavorable prognostic factors. These findings provide potential therapeutic targets for LMS, particularly in combination with immune checkpoint inhibitors. Further investigation into the molecular mechanisms of m6A-mediated regulation of PD-L1 and c-Myc expression is required to develop more effective treatments for LMS.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104221"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760449","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}
A disintegrin and metalloproteinase 28 (ADAM28), which comprises membrane-anchored form (ADAM28m) and short-secreted form (ADAM28s), is overexpressed by carcinoma cells and involved in cancer cell proliferation and metastasis in several cancers. However, little is known about the implications of ADAM28 in esophageal squamous cell carcinoma (ESCC). In this study, we investigated the expression and clinical implication of ADAM28 in ESCC and examined the molecular mechanism of ADAM28-mediated ESCC cell proliferation. Immunoblotting analysis demonstrated that ADAM28s is overexpressed in active forms of 42 and/or 40 kDa in ESCC tissues compared with nonneoplastic esophageal tissues. Immunohistochemistry and deep learning artificial intelligence showed that ADAM28s is expressed mainly by carcinoma cells in the ESCC tissue, and the 5-year overall survival and disease-specific survival rates in cases with extensive immunostaining are significantly worse than those in low immunostaining cases. Among several factors examined, interleukin 6 (IL-6) enhanced ADAM28s expression in ESCC cell lines (TE-1 and KYSE-140), which exhibited ADAM28 expression but not in a cell line without the expression (TE-8). Proliferation of TE-1 and KYSE-140 cells under IL-6 stimulation was effectively inhibited by treatment with anti-ADAM28 antibody or siRNA-mediated downregulation of ADAM28, whereas no such effect was observed in TE-8 cells. In mouse ESCC cell xenografts, tumor growth of KYSE-140ffLuc-cp156 cells was significantly reduced by treatment with the anti-ADAM28 antibody compared with the control immunoglobulin G–treated group. These results show that ADAM28s is overexpressed as active forms in ESCC cells and suggest that ADAM28s is involved in cell proliferation probably through the IL-6 signaling pathway.
{"title":"Increased Expression of Secreted Form A Disintegrin and Metalloproteinase 28 (ADAM28s) in Esophageal Squamous Cell Carcinoma: Implication for Carcinoma Cell Proliferation via Interleukin 6 Receptor Shedding","authors":"Keita Kouzu , Satsuki Mochizuki , Hironori Tsujimoto , Yusuke Ishibashi , Ines P. Nearchou , Kentaro Ohara , Seiichiro Fujishima , Yoji Kishi , Susumu Matsukuma , Yasunori Okada , Hideki Ueno","doi":"10.1016/j.labinv.2025.104222","DOIUrl":"10.1016/j.labinv.2025.104222","url":null,"abstract":"<div><div>A disintegrin and metalloproteinase 28 (ADAM28), which comprises membrane-anchored form (ADAM28m) and short-secreted form (ADAM28s), is overexpressed by carcinoma cells and involved in cancer cell proliferation and metastasis in several cancers. However, little is known about the implications of ADAM28 in esophageal squamous cell carcinoma (ESCC). In this study, we investigated the expression and clinical implication of ADAM28 in ESCC and examined the molecular mechanism of ADAM28-mediated ESCC cell proliferation. Immunoblotting analysis demonstrated that ADAM28s is overexpressed in active forms of 42 and/or 40 kDa in ESCC tissues compared with nonneoplastic esophageal tissues. Immunohistochemistry and deep learning artificial intelligence showed that ADAM28s is expressed mainly by carcinoma cells in the ESCC tissue, and the 5-year overall survival and disease-specific survival rates in cases with extensive immunostaining are significantly worse than those in low immunostaining cases. Among several factors examined, interleukin 6 (IL-6) enhanced ADAM28s expression in ESCC cell lines (TE-1 and KYSE-140), which exhibited ADAM28 expression but not in a cell line without the expression (TE-8). Proliferation of TE-1 and KYSE-140 cells under IL-6 stimulation was effectively inhibited by treatment with anti-ADAM28 antibody or siRNA-mediated downregulation of <em>ADAM28</em>, whereas no such effect was observed in TE-8 cells. In mouse ESCC cell xenografts, tumor growth of KYSE-140<sup>ffLuc-cp156</sup> cells was significantly reduced by treatment with the anti-ADAM28 antibody compared with the control immunoglobulin G–treated group. These results show that ADAM28s is overexpressed as active forms in ESCC cells and suggest that ADAM28s is involved in cell proliferation probably through the IL-6 signaling pathway.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104222"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760450","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}
Pub Date : 2025-11-01Epub Date: 2025-07-29DOI: 10.1016/j.labinv.2025.104223
Aihua Guo , Yilin Yu , Qinpeng Guo , Enhuan Zhang , Huaqin Lin , Mei Feng , Peilin Zhong , Jie Lin , Linghua Wang , Xiurong Lin , Haixia Wu , Yang Sun
This study aimed to identify and characterize novel macrophage–related molecular mechanisms underlying immunosuppression and tumor progression in cervical cancer. Through a systematic integrative analysis guided by immune-related gene signatures and robust regression modeling, we identified PARVB as a novel macrophage–associated prognostic gene with strong predictive value across multiple data sets. Further validation using large-scale transcriptomic data and single-cell RNA-sequencing profiles revealed that PARVB likely activates the SMAD signaling axis, leading to the upregulation of TNFSF13, a key driver of M2 macrophage polarization. This PARVB-SMAD3-TNFSF13 axis enhances interactions between M2 macrophages and TNFSF13+ subsets, promoting regulatory T-cell induction and fostering an immunosuppressive tumor microenvironment. Functional assays and multiplex immunohistochemistry further confirmed that this axis drives tumor proliferation and immune evasion. Collectively, our findings uncover a critical PARVB-driven signaling cascade that reprograms macrophages into an immunosuppressive M2 phenotype, facilitating immune escape and cervical cancer progression. Targeting this axis presents a promising therapeutic strategy to reshape the tumor microenvironment and improve immunotherapeutic outcomes.
{"title":"Potential Role of PARVB in Macrophage-Mediated Immunosuppression and Cervical Cancer Progression","authors":"Aihua Guo , Yilin Yu , Qinpeng Guo , Enhuan Zhang , Huaqin Lin , Mei Feng , Peilin Zhong , Jie Lin , Linghua Wang , Xiurong Lin , Haixia Wu , Yang Sun","doi":"10.1016/j.labinv.2025.104223","DOIUrl":"10.1016/j.labinv.2025.104223","url":null,"abstract":"<div><div>This study aimed to identify and characterize novel macrophage–related molecular mechanisms underlying immunosuppression and tumor progression in cervical cancer. Through a systematic integrative analysis guided by immune-related gene signatures and robust regression modeling, we identified PARVB as a novel macrophage–associated prognostic gene with strong predictive value across multiple data sets. Further validation using large-scale transcriptomic data and single-cell RNA-sequencing profiles revealed that PARVB likely activates the SMAD signaling axis, leading to the upregulation of TNFSF13, a key driver of M2 macrophage polarization. This PARVB-SMAD3-TNFSF13 axis enhances interactions between M2 macrophages and TNFSF13<sup>+</sup> subsets, promoting regulatory T-cell induction and fostering an immunosuppressive tumor microenvironment. Functional assays and multiplex immunohistochemistry further confirmed that this axis drives tumor proliferation and immune evasion. Collectively, our findings uncover a critical PARVB-driven signaling cascade that reprograms macrophages into an immunosuppressive M2 phenotype, facilitating immune escape and cervical cancer progression. Targeting this axis presents a promising therapeutic strategy to reshape the tumor microenvironment and improve immunotherapeutic outcomes.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 11","pages":"Article 104223"},"PeriodicalIF":4.2,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144760482","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}
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