Pub Date : 2025-09-16DOI: 10.1016/j.labinv.2025.104242
Margaret Tulessin , Ludwig Beer , Stephanie Roessler , Anika Beckers , Darko Castven , Diego Francesco Calvisi , Xin Chen , Sebastian Lange , Nicole Pfarr , Jens Marquardt , Theresa Hildegard Wirtz , Marie-Luise Berres , Katja Steiger , Tanja Groll , Carolin Mogler
Cholangiocarcinoma (CCA) is an aggressive malignancy that originates in the bile ducts and is characterized by late-stage diagnosis and limited treatment options. CCA accounts for approximately 10% to 15% of primary liver tumors. Recent genetic studies have shed new light on this disease, exploring CCA’s complexity and finding more effective treatment strategies, particularly based on identifying actionable mutations. Various mouse models for CCA have been established; however, the extent to which these models reflect the complexity of human is not well investigated. Therefore, this study aimed to characterize the available mouse models for CCA studies and compare their characteristics, advantages, and challenges in resemblance to human CCA. We applied tissue-based techniques using classical hematoxylin and eosin, Sirius red, and immunohistochemistry of 16 markers for in-depth characterization of tumor cells, and the tumor microenvironment of 11 different mouse models. Our findings demonstrate that CCAs present with various tumor subtypes, tumor growth patterns, morphologic subtypes, and tumor microenvironment activity. Furthermore, we report here that neoplastic lesions other than CCA, such as hepatocellular carcinoma and nonneoplastic changes in the liver parenchyma (eg, steatosis), occur with significant differences among the investigated models. Nine out of 11 investigated models were suitable for CCA studies as they resemble human CCA features. Overall, our data show that mouse models of CCA represent a valid tool to investigate this deadly disease, but they should be carefully selected, depending on the study’s aims and targets in advance.
{"title":"Detailed Characterization and Comparison of Mouse Models for Cholangiocarcinoma","authors":"Margaret Tulessin , Ludwig Beer , Stephanie Roessler , Anika Beckers , Darko Castven , Diego Francesco Calvisi , Xin Chen , Sebastian Lange , Nicole Pfarr , Jens Marquardt , Theresa Hildegard Wirtz , Marie-Luise Berres , Katja Steiger , Tanja Groll , Carolin Mogler","doi":"10.1016/j.labinv.2025.104242","DOIUrl":"10.1016/j.labinv.2025.104242","url":null,"abstract":"<div><div>Cholangiocarcinoma (CCA) is an aggressive malignancy that originates in the bile ducts and is characterized by late-stage diagnosis and limited treatment options. CCA accounts for approximately 10% to 15% of primary liver tumors. Recent genetic studies have shed new light on this disease, exploring CCA’s complexity and finding more effective treatment strategies, particularly based on identifying actionable mutations. Various mouse models for CCA have been established; however, the extent to which these models reflect the complexity of human is not well investigated. Therefore, this study aimed to characterize the available mouse models for CCA studies and compare their characteristics, advantages, and challenges in resemblance to human CCA. We applied tissue-based techniques using classical hematoxylin and eosin, Sirius red, and immunohistochemistry of 16 markers for in-depth characterization of tumor cells, and the tumor microenvironment of 11 different mouse models. Our findings demonstrate that CCAs present with various tumor subtypes, tumor growth patterns, morphologic subtypes, and tumor microenvironment activity. Furthermore, we report here that neoplastic lesions other than CCA, such as hepatocellular carcinoma and nonneoplastic changes in the liver parenchyma (eg, steatosis), occur with significant differences among the investigated models. Nine out of 11 investigated models were suitable for CCA studies as they resemble human CCA features. Overall, our data show that mouse models of CCA represent a valid tool to investigate this deadly disease, but they should be carefully selected, depending on the study’s aims and targets in advance.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104242"},"PeriodicalIF":4.2,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145086168","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 : 2025-09-15DOI: 10.1016/j.labinv.2025.104240
Ekaterina Menshikova , Kristin Deeb , Elizabeth M. Genega , Krisztina Hanley , Gulisa Turashvili
Targeted therapy directed against human epidermal growth factor receptor 2 (HER2) has shown promising results in HER2-positive endometrial cancer. Recent limited data suggest significant intratumoral heterogeneity in HER2 expression in serous carcinomas. We aimed to evaluate HER2 heterogeneity at the protein and gene levels and the impact of variable definitions in endometrial carcinomas including nonserous subtypes. We retrospectively identified biopsies and surgical specimens with available HER2 immunohistochemical (IHC) stains and fluorescence in situ hybridization (FISH) results. IHC stains and FISH data were reevaluated to assess variability in the HER2 protein expression and gene amplification. The overall HER2-positivity rate was 31% using the endometrial criteria, and 42.6% by the gastric criteria. Heterogeneous IHC staining was observed in 45.7% of tumors, predominating in 2+/3+ scores (P < .001). Re-evaluation of FISH revealed a 31% rate of heterogeneous gene amplification. Our study demonstrates a high frequency of HER2 heterogeneity at both the protein and the gene levels. Further studies on HER2 heterogeneity and treatment response are warranted for refining standardized testing and reporting algorithms.
{"title":"Analysis of Human Epidermal Growth Factor Receptor 2 (HER2) Heterogeneity at the Protein and Gene Levels in Endometrial Cancer: Refining HER2 Reporting and HER2-Directed Therapies","authors":"Ekaterina Menshikova , Kristin Deeb , Elizabeth M. Genega , Krisztina Hanley , Gulisa Turashvili","doi":"10.1016/j.labinv.2025.104240","DOIUrl":"10.1016/j.labinv.2025.104240","url":null,"abstract":"<div><div>Targeted therapy directed against human epidermal growth factor receptor 2 (HER2) has shown promising results in HER2-positive endometrial cancer. Recent limited data suggest significant intratumoral heterogeneity in HER2 expression in serous carcinomas. We aimed to evaluate HER2 heterogeneity at the protein and gene levels and the impact of variable definitions in endometrial carcinomas including nonserous subtypes. We retrospectively identified biopsies and surgical specimens with available HER2 immunohistochemical (IHC) stains and fluorescence in situ hybridization (FISH) results. IHC stains and FISH data were reevaluated to assess variability in the HER2 protein expression and gene amplification. The overall HER2-positivity rate was 31% using the endometrial criteria, and 42.6% by the gastric criteria. Heterogeneous IHC staining was observed in 45.7% of tumors, predominating in 2+/3+ scores (<em>P</em> < .001). Re-evaluation of FISH revealed a 31% rate of heterogeneous gene amplification. Our study demonstrates a high frequency of HER2 heterogeneity at both the protein and the gene levels. Further studies on HER2 heterogeneity and treatment response are warranted for refining standardized testing and reporting algorithms.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104240"},"PeriodicalIF":4.2,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145081173","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 : 2025-09-12DOI: 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":"2025-09-12","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}
Pub Date : 2025-09-04DOI: 10.1016/j.labinv.2025.104238
Dong Ren , Chenchen Niu , Nyein Htun , Xin Zhang , Jiadi He , Ronggang Li , Qiongru Liu , Vincent Lee , Robert A. Edwards , Grace G. Zhou , Brandon M. Lehrich , Derek H. Liu , Angie Nguyen , Jeff Chan , Nicholas R. Pannunzio , Edward C. Kuan , Beverly Y. Wang
Sinonasal mucosal melanoma (SNMM) is a rare aggressive malignancy of the sinonasal tract. Due to its advanced clinical presentation and frequent late-stage diagnosis, the 5-year survival rate is <30%, with an even worse prognosis in patients with distant metastasis (SNMM-M). Therefore, characterizing the molecular landscape of SNMM may provide novel therapeutic targets for SNMM-M. This study aimed to decipher the histopathological and molecular landscape of SNMM-M and yields novel insights into potential therapeutic approaches using targeted DNA and RNA next-generation sequencing, immunohistochemistry, and fluorescence in situ hybridization. SNMM-M cases were characterized by epithelioid-predominant morphology, frequent tumor necrosis, minimal pleomorphism, and sparse tumor-infiltrating lymphocytes (TILs). A significant association between the absence of brisk TILs and metastasis in SNMM was noted. Moreover, the presence of lymphovascular invasion and absence of brisk TILs were both significantly associated with poorer overall survival in patients with SNMM. Both DNA and RNA sequencing identified no gene fusions, whereas DNA sequencing revealed 304 genomic alterations across 186 genes, including 9 multihit genes. Notably, missense mutations in structure-specific endonuclease subunit (SLX4), a key homologous recombination repair scaffold protein, were exclusively detected in SNMM-M, and patients with SLX4 mutations exhibited significantly worse survival (median, 9.9 vs 41.2 months without SLX4 mutations; P < .0001). A comparative analysis of genomic alterations and copy number variations of clinically actionable genes between SNMM-M and SNMM without distant metastasis (SNMM-nM) revealed that CDK4 gains/amplifications were commonly seen in SNMM-M cases, whereas receptor tyrosine kinase and homologous recombination repair gene alterations were highly enriched in SNMM-nM cases. Additionally, PD-L1 was more commonly expressed in SNMM-nM. These exploratory findings suggest that SLX4 mutation may serve as a potential prognostic biomarker and that CDK4 inhibitors could represent promising therapeutic options for SNMM-M. However, given the limited sample size, further validation in larger studies is essential to confirm these findings.
{"title":"Exploring Molecular Characteristics and Therapeutic Strategies for Primary Sinonasal Mucosal Melanoma With Distant Metastasis","authors":"Dong Ren , Chenchen Niu , Nyein Htun , Xin Zhang , Jiadi He , Ronggang Li , Qiongru Liu , Vincent Lee , Robert A. Edwards , Grace G. Zhou , Brandon M. Lehrich , Derek H. Liu , Angie Nguyen , Jeff Chan , Nicholas R. Pannunzio , Edward C. Kuan , Beverly Y. Wang","doi":"10.1016/j.labinv.2025.104238","DOIUrl":"10.1016/j.labinv.2025.104238","url":null,"abstract":"<div><div>Sinonasal mucosal melanoma (SNMM) is a rare aggressive malignancy of the sinonasal tract. Due to its advanced clinical presentation and frequent late-stage diagnosis, the 5-year survival rate is <30%, with an even worse prognosis in patients with distant metastasis (SNMM-M). Therefore, characterizing the molecular landscape of SNMM may provide novel therapeutic targets for SNMM-M. This study aimed to decipher the histopathological and molecular landscape of SNMM-M and yields novel insights into potential therapeutic approaches using targeted DNA and RNA next-generation sequencing, immunohistochemistry, and fluorescence in situ hybridization. SNMM-M cases were characterized by epithelioid-predominant morphology, frequent tumor necrosis, minimal pleomorphism, and sparse tumor-infiltrating lymphocytes (TILs). A significant association between the absence of brisk TILs and metastasis in SNMM was noted. Moreover, the presence of lymphovascular invasion and absence of brisk TILs were both significantly associated with poorer overall survival in patients with SNMM. Both DNA and RNA sequencing identified no gene fusions, whereas DNA sequencing revealed 304 genomic alterations across 186 genes, including 9 multihit genes. Notably, missense mutations in structure-specific endonuclease subunit (<em>SLX4</em>), a key homologous recombination repair scaffold protein, were exclusively detected in SNMM-M, and patients with <em>SLX4</em> mutations exhibited significantly worse survival (median, 9.9 vs 41.2 months without <em>SLX4</em> mutations; <em>P</em> < .0001). A comparative analysis of genomic alterations and copy number variations of clinically actionable genes between SNMM-M and SNMM without distant metastasis (SNMM-nM) revealed that <em>CDK4</em> gains/amplifications were commonly seen in SNMM-M cases, whereas receptor tyrosine kinase and homologous recombination repair gene alterations were highly enriched in SNMM-nM cases. Additionally, PD-L1 was more commonly expressed in SNMM-nM. These exploratory findings suggest that <em>SLX4</em> mutation may serve as a potential prognostic biomarker and that CDK4 inhibitors could represent promising therapeutic options for SNMM-M. However, given the limited sample size, further validation in larger studies is essential to confirm these findings.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104238"},"PeriodicalIF":4.2,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145008311","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}
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 TNC 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 TNC knockdown suppressed it. Spatial transcriptomics revealed elevated TNC 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.
{"title":"Tenascin-C Expression in Relation to Tumor-Stroma Interaction in Ameloblastoma","authors":"Satoko Sumi , Shohei Yoshimoto , Kanako Suyama , Masahide Taguchi , Hiromitsu Morita , Akimitsu Hiraki , 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":"2025-09-03","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}
Pub 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":"2025-09-02","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 : 2025-09-02DOI: 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":"2025-09-02","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 : 2025-09-01DOI: 10.1016/j.labinv.2025.104234
Frank W.J. Heijboer , Jules L. Derks , Dana A.M. Mustafa , Nicole Rijnsburger , Bregtje C.M. Hermans , Lisa M. Hillen , PALGA-Group , Ernst-Jan M. Speel , Anne-Marie C. Dingemans , Jan H. von der Thüsen
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 >10 was considered positive (+), and >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: NEUROD1high-ASCL1high (10%), ASCL1high (22%), POU2F3high (5%), YAP1high (11%), and NAPYlow (51%). Both ASCL1high subgroups correlated with DLL3high and high neuroendocrine (NE) marker expression. YAP1high was enriched for pRb+. POU2F3high and YAP1high subgroups were NE marker low and DLL3low. GeoMX Digital Spatial Profiling identified 4 upregulated genes involved in immune system and/or tumor development in the NEUROD1high-ASCL1high-POU2F3high- group. In this comprehensive evaluation of NAPY markers in LCNEC, we observed 5 expression-based subgroups: NEUROD1high-ASCL1high, ASCL1high, POU2F3high, YAP1high, and NAPYlow. The NE subgroups (NEUROD1high-ASCL1high and ASCL1high) were recognized with DLL3high expression. Compared with the proportion known in SCLC, more NAPYlow and YAP1high and fewer POU2F3high 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.
{"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 , Jules L. Derks , Dana A.M. Mustafa , Nicole Rijnsburger , Bregtje C.M. Hermans , Lisa M. Hillen , PALGA-Group , Ernst-Jan M. Speel , Anne-Marie C. Dingemans , 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 >10 was considered positive (+), and >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<sup>high</sup>-ASCL1<sup>high</sup> (10%), ASCL1<sup>high</sup> (22%), POU2F3<sup>high</sup> (5%), YAP1<sup>high</sup> (11%), and NAPY<sup>low</sup> (51%). Both ASCL1<sup>high</sup> subgroups correlated with DLL3<sup>high</sup> and high neuroendocrine (NE) marker expression. YAP1<sup>high</sup> was enriched for pRb+. POU2F3<sup>high</sup> and YAP1<sup>high</sup> subgroups were NE marker low and DLL3<sup>low</sup>. GeoMX Digital Spatial Profiling identified 4 upregulated genes involved in immune system and/or tumor development in the NEUROD1<sup>high</sup>-ASCL1<sup>high</sup>-POU2F3<sup>high</sup>- group. In this comprehensive evaluation of NAPY markers in LCNEC, we observed 5 expression-based subgroups: NEUROD1<sup>high</sup>-ASCL1<sup>high</sup>, ASCL1<sup>high</sup>, POU2F3<sup>high</sup>, YAP1<sup>high</sup>, and NAPY<sup>low</sup>. The NE subgroups (NEUROD1<sup>high</sup>-ASCL1<sup>high</sup> and ASCL1<sup>high</sup>) were recognized with DLL3<sup>high</sup> expression. Compared with the proportion known in SCLC, more NAPY<sup>low</sup> and YAP1<sup>high</sup> and fewer POU2F3<sup>high</sup> 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":"2025-09-01","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}
Pub Date : 2025-08-27DOI: 10.1016/j.labinv.2025.104233
Nay N.N. Chan , Patricia Gaule , Julia Benanto , Liam Scott , Charles J. Robbins , Mengni He , Katherine Bates , Revekka Khaimova , Daniel C. Liebler , Regan Fulton , David L. Rimm
The recent approval of antibody-drug conjugates targeting human epidermal growth factor receptor 2 (HER2) (such as trastuzumab deruxtecan [T-DXd]) has led to challenges for the immunohistochemical (IHC) companion diagnostic test because the test was optimized for gene-amplified levels of HER2. Here, we develop and validate an objective test for low-level HER2 expression toward more accurate selection of patients for T-DXd. We validated the high-sensitivity HER2 assay using a mix of the requirements for an IHC assay and that of a ligand-binding assay. Then, we prospectively tested it on 316 core biopsy specimens received by the Yale Pathology Laboratories from August 2022 to August 2023. Using a 40-case breast cancer tissue validation set, we find very high accuracy and precision with a coefficient of variation <10% and define a reportable range for the assay in attomoles per square millimeter. These prospective cases not only show the dynamic range of HER2 expression but also the discordance of Yale Pathology Labs staff pathologist scores with quantitative measurements, especially in the low range of HER2. We find that 6% of the cohort was below the limit of detection of this more sensitive assay, whereas 71% of the IHC 0 cases were above the limit of quantification. Efforts are underway to determine a possible threshold expression level required for T-DXd response. In summary, this assay validation study provides a method for accurate, objective measurement of HER2 and has the potential to improve selection of patients for T-DXd or similarly targeted antibody-drug conjugate therapies in future.
{"title":"Validation and Prospective Testing of a High-Sensitivity, Quantitative Analytic Assay for HER2 on Histopathology Slides","authors":"Nay N.N. Chan , Patricia Gaule , Julia Benanto , Liam Scott , Charles J. Robbins , Mengni He , Katherine Bates , Revekka Khaimova , Daniel C. Liebler , Regan Fulton , David L. Rimm","doi":"10.1016/j.labinv.2025.104233","DOIUrl":"10.1016/j.labinv.2025.104233","url":null,"abstract":"<div><div>The recent approval of antibody-drug conjugates targeting human epidermal growth factor receptor 2 (HER2) (such as trastuzumab deruxtecan [T-DXd]) has led to challenges for the immunohistochemical (IHC) companion diagnostic test because the test was optimized for gene-amplified levels of HER2. Here, we develop and validate an objective test for low-level HER2 expression toward more accurate selection of patients for T-DXd. We validated the high-sensitivity HER2 assay using a mix of the requirements for an IHC assay and that of a ligand-binding assay. Then, we prospectively tested it on 316 core biopsy specimens received by the Yale Pathology Laboratories from August 2022 to August 2023. Using a 40-case breast cancer tissue validation set, we find very high accuracy and precision with a coefficient of variation <10% and define a reportable range for the assay in attomoles per square millimeter. These prospective cases not only show the dynamic range of HER2 expression but also the discordance of Yale Pathology Labs staff pathologist scores with quantitative measurements, especially in the low range of HER2. We find that 6% of the cohort was below the limit of detection of this more sensitive assay, whereas 71% of the IHC 0 cases were above the limit of quantification. Efforts are underway to determine a possible threshold expression level required for T-DXd response. In summary, this assay validation study provides a method for accurate, objective measurement of HER2 and has the potential to improve selection of patients for T-DXd or similarly targeted antibody-drug conjugate therapies in future.</div></div>","PeriodicalId":17930,"journal":{"name":"Laboratory Investigation","volume":"105 12","pages":"Article 104233"},"PeriodicalIF":4.2,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144959187","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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