Maaike Anna Hempenius, Maran A Eenkhoorn, Henrik Høeg, David J Dabbs, Bert van der Vegt, Seshi R Sompuram, Nils A ‘t Hart
Aims
Recently, human epidermal growth factor 2 (HER2)-low (i.e. HER2 score 1+ or 2+ without amplification) breast cancer patients became eligible for trastuzumab–deruxtecan treatment. To improve assay standardisation and detection of HER2-low in a quantitative manner, we conducted an external quality assessment-like study in the Netherlands. Dynamic range cell lines and immunohistochemistry (IHC) calibrators were used to quantify HER2 expression and to assess interlaboratory variability.
Methods and results
Three blank slides with a dynamic range cell line and an IHC calibrator were stained with routine HER2 assays by 35 laboratories. Four different antibody clones were used: 19 (54.3%) 4B5, six (17.1%) A0485, five (14.3%) DG44 (HercepTest) and five (14.3%) SP3. Laboratories used two different detection kits for 4B5 assays: 14 (73.7%) ultraView and five (26.3%) OptiView. Variability of HER2 expression in cell lines, measured with artificial intelligence software, was median (min–max) = negative core 0.5% (0.0–57.0), 1+ core 4.3% (1.6–71.3), 2+ core 42.8% (30.4–92.6) and 3+ core 96.2% (91.8–98.8). The calibrators DG44 and 4B5 OptiView had the highest analytical sensitivity, closely followed by 4B5 ultraView. SP3 was the least sensitive. Calibrators of A0485 assays were not analysable due to background staining.
Conclusions
As assays were validated for detecting HER2-amplified tumours, not all assays and antibodies proved suitable for HER2-low detection. Some tests showed distinct expression in the negative cell line. Dynamic range cell line controls and quantitative analysis using calibrators demonstrated more interlaboratory variability than commonly appreciated. Revalidation of HER2 tests by laboratories is needed to ensure clinical applicable HER2-low assays.
{"title":"Quantitative comparison of immunohistochemical HER2-low detection in an interlaboratory study","authors":"Maaike Anna Hempenius, Maran A Eenkhoorn, Henrik Høeg, David J Dabbs, Bert van der Vegt, Seshi R Sompuram, Nils A ‘t Hart","doi":"10.1111/his.15273","DOIUrl":"10.1111/his.15273","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Aims</h3>\u0000 \u0000 <p>Recently, human epidermal growth factor 2 (HER2)-low (i.e. HER2 score 1+ or 2+ without amplification) breast cancer patients became eligible for trastuzumab–deruxtecan treatment. To improve assay standardisation and detection of HER2-low in a quantitative manner, we conducted an external quality assessment-like study in the Netherlands. Dynamic range cell lines and immunohistochemistry (IHC) calibrators were used to quantify HER2 expression and to assess interlaboratory variability.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods and results</h3>\u0000 \u0000 <p>Three blank slides with a dynamic range cell line and an IHC calibrator were stained with routine HER2 assays by 35 laboratories. Four different antibody clones were used: 19 (54.3%) 4B5, six (17.1%) A0485, five (14.3%) DG44 (HercepTest) and five (14.3%) SP3. Laboratories used two different detection kits for 4B5 assays: 14 (73.7%) ultraView and five (26.3%) OptiView. Variability of HER2 expression in cell lines, measured with artificial intelligence software, was median (min–max) = negative core 0.5% (0.0–57.0), 1+ core 4.3% (1.6–71.3), 2+ core 42.8% (30.4–92.6) and 3+ core 96.2% (91.8–98.8). The calibrators DG44 and 4B5 OptiView had the highest analytical sensitivity, closely followed by 4B5 ultraView. SP3 was the least sensitive. Calibrators of A0485 assays were not analysable due to background staining.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>As assays were validated for detecting HER2-amplified tumours, not all assays and antibodies proved suitable for HER2-low detection. Some tests showed distinct expression in the negative cell line. Dynamic range cell line controls and quantitative analysis using calibrators demonstrated more interlaboratory variability than commonly appreciated. Revalidation of HER2 tests by laboratories is needed to ensure clinical applicable HER2-low assays.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13219,"journal":{"name":"Histopathology","volume":"85 6","pages":"920-928"},"PeriodicalIF":3.9,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/his.15273","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141792355","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}
Anandi Lobo, Katrina Collins, Seema Kaushal, Andres M Acosta, Mahmut Akgul, Amit K Adhya, Hikmat A Al-Ahmadie, Khaleel I Al-Obaidy, Ali Amin, Mahul B Amin, Manju Aron, Bonnie L Balzer, Rupanita Biswal, Subashish Mohanty, Lisa Browning, Indranil Chakrabarti, Luca Cima, Alessia Cimadamore, Sangeeta Desai, Jasreman Dhillon, Akansha Deshwal, Guillermo G Diego, Preeti Diwaker, Laurence A Galea, Cristina Magi-Galluzzi, Giovanna A Giannico, Nilesh S Gupta, Aiman Haider, Michelle S Hirsch, Kenneth A Iczkowski, Samriti Arora, Ekta Jain, Deepika Jain, Shilpy Jha, Shivani Kandukuri, Chia-Sui Kao, Sunny, Oleksandr N Kryvenko, Ramani M Kumar, Niraj Kumari, Lakshmi P Kunju, Levente Kuthi, João Lobo, Jose I Lopez, Daniel J Luthringer, Fiona Maclean, Claudia Manini, Rahul Mannan, María G Martos, Rohit Mehra, Santosh Menon, Pritinanda Mishra, Holger Moch, Rodolfo Montironi, Manas R Baisakh, George J Netto, Lovelesh K Nigam, Adeboye O Osunkoya, Francesca Pagliuca, Gladell P Paner, Angel Panizo, Anil V Parwani, Maria M Picken, Susan Prendeville, Christopher G Przybycin, Suvendu Purkait, Francisco J Queipo, B Vishal Rao, Priya Rao, Victor E Reuter, Sankalp Sancheti, Ankur R Sangoi, Rohan Sardana, Swati Satturwar, Rajal B Shah, Shivani Sharma, Mallika Dixit, Monica Verma, Deepika Sirohi, Steven C Smith, Shailesh Soni, Sandhya Sundaram, Meenakshi Swain, Maria Tretiakova, Kiril Trpkov, Gorka MuñizUnamunzaga, Ming Zhou, Sean R Williamson, Antonio Lopez-Beltran, Liang Cheng, Sambit K Mohanty
<div> <section> <h3> Aims</h3> <p>Urothelial carcinoma (UC) demonstrates significant molecular and histologic heterogeneity. The WHO 2022 classification has hinted at adding molecular signatures to the morphologic diagnosis. As morphology and associated molecular repertoire may potentially translate to choices of and response to therapy and relapse rate, broader acceptability of recognizing these key features among uropathologists is needed. This prompted an international survey to ascertain the practice patterns in classical/subtype UC among uropathologists across the globe.</p> </section> <section> <h3> Methods and Results</h3> <p>A survey instrument was shared among 98 uropathologists using SurveyMonkey software. Anonymized respondent data were analysed. The response rate was 85%. A majority were in concordance with the profiles of luminal (93%) and basal (82%) types. Opinion on the <i>FGFR3</i> testing platform was variable. While 95% concurred that <i>TERT</i> promoter mutation is the key driver in UC, 72% had the opinion that <i>APOBEC</i> mutagenesis is the main signature in muscle invasive bladder cancer (MIBC). Uropathologists have divergent opinions on MIBC and <i>ERCC2</i> mutations. Among the participants, 94% would quantify aggressive micropapillary and sarcomatoid histology, while 88% would reevaluate another transurethral resection of the bladder tumour specimen in nonmuscle invasive tumour with micropapillary, small cell, or sarcomatoid histology. A leading number agreed to specific molecular signatures of micropapillary (93%), plasmacytoid (97%), and small cell (86%) subtypes. Ninety-six percent of participants agreed that a small-cell component portends a more aggressive course and should be treated with neoadjuvant chemotherapy and 63% would perform <i>HER2/neu</i> testing only on oncologist's request in advanced tumours. Ninety percent agreed that microsatellite instability testing, although not a standard protocol, should be considered in young patients with upper tract UC. Eighty-six percent agreed that UC with high tumour mutational burden would be a better candidate for immunotherapy.</p> </section> <section> <h3> Conclusion</h3> <p>In the era of precision medicine, enhanced understanding of molecular heterogeneity of UC will contribute to better therapeutic options, novel biomarker discovery, innovative management protocols, and outcomes. Our survey provides a broad perspective of pathologists' perceptions and experience regarding incorporation of histomolecular approaches to “personalize” therapy. Due to variable clinical adoption, there is a need for additional data using uniform study criteria. This will drive generation of best practice guidelines in this area for widespread and consistent c
{"title":"Advances, recognition, and interpretation of molecular heterogeneity among conventional and subtype histology of urothelial carcinoma (UC): a survey among urologic pathologists and comprehensive review of the literature","authors":"Anandi Lobo, Katrina Collins, Seema Kaushal, Andres M Acosta, Mahmut Akgul, Amit K Adhya, Hikmat A Al-Ahmadie, Khaleel I Al-Obaidy, Ali Amin, Mahul B Amin, Manju Aron, Bonnie L Balzer, Rupanita Biswal, Subashish Mohanty, Lisa Browning, Indranil Chakrabarti, Luca Cima, Alessia Cimadamore, Sangeeta Desai, Jasreman Dhillon, Akansha Deshwal, Guillermo G Diego, Preeti Diwaker, Laurence A Galea, Cristina Magi-Galluzzi, Giovanna A Giannico, Nilesh S Gupta, Aiman Haider, Michelle S Hirsch, Kenneth A Iczkowski, Samriti Arora, Ekta Jain, Deepika Jain, Shilpy Jha, Shivani Kandukuri, Chia-Sui Kao, Sunny, Oleksandr N Kryvenko, Ramani M Kumar, Niraj Kumari, Lakshmi P Kunju, Levente Kuthi, João Lobo, Jose I Lopez, Daniel J Luthringer, Fiona Maclean, Claudia Manini, Rahul Mannan, María G Martos, Rohit Mehra, Santosh Menon, Pritinanda Mishra, Holger Moch, Rodolfo Montironi, Manas R Baisakh, George J Netto, Lovelesh K Nigam, Adeboye O Osunkoya, Francesca Pagliuca, Gladell P Paner, Angel Panizo, Anil V Parwani, Maria M Picken, Susan Prendeville, Christopher G Przybycin, Suvendu Purkait, Francisco J Queipo, B Vishal Rao, Priya Rao, Victor E Reuter, Sankalp Sancheti, Ankur R Sangoi, Rohan Sardana, Swati Satturwar, Rajal B Shah, Shivani Sharma, Mallika Dixit, Monica Verma, Deepika Sirohi, Steven C Smith, Shailesh Soni, Sandhya Sundaram, Meenakshi Swain, Maria Tretiakova, Kiril Trpkov, Gorka MuñizUnamunzaga, Ming Zhou, Sean R Williamson, Antonio Lopez-Beltran, Liang Cheng, Sambit K Mohanty","doi":"10.1111/his.15287","DOIUrl":"10.1111/his.15287","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Aims</h3>\u0000 \u0000 <p>Urothelial carcinoma (UC) demonstrates significant molecular and histologic heterogeneity. The WHO 2022 classification has hinted at adding molecular signatures to the morphologic diagnosis. As morphology and associated molecular repertoire may potentially translate to choices of and response to therapy and relapse rate, broader acceptability of recognizing these key features among uropathologists is needed. This prompted an international survey to ascertain the practice patterns in classical/subtype UC among uropathologists across the globe.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods and Results</h3>\u0000 \u0000 <p>A survey instrument was shared among 98 uropathologists using SurveyMonkey software. Anonymized respondent data were analysed. The response rate was 85%. A majority were in concordance with the profiles of luminal (93%) and basal (82%) types. Opinion on the <i>FGFR3</i> testing platform was variable. While 95% concurred that <i>TERT</i> promoter mutation is the key driver in UC, 72% had the opinion that <i>APOBEC</i> mutagenesis is the main signature in muscle invasive bladder cancer (MIBC). Uropathologists have divergent opinions on MIBC and <i>ERCC2</i> mutations. Among the participants, 94% would quantify aggressive micropapillary and sarcomatoid histology, while 88% would reevaluate another transurethral resection of the bladder tumour specimen in nonmuscle invasive tumour with micropapillary, small cell, or sarcomatoid histology. A leading number agreed to specific molecular signatures of micropapillary (93%), plasmacytoid (97%), and small cell (86%) subtypes. Ninety-six percent of participants agreed that a small-cell component portends a more aggressive course and should be treated with neoadjuvant chemotherapy and 63% would perform <i>HER2/neu</i> testing only on oncologist's request in advanced tumours. Ninety percent agreed that microsatellite instability testing, although not a standard protocol, should be considered in young patients with upper tract UC. Eighty-six percent agreed that UC with high tumour mutational burden would be a better candidate for immunotherapy.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>In the era of precision medicine, enhanced understanding of molecular heterogeneity of UC will contribute to better therapeutic options, novel biomarker discovery, innovative management protocols, and outcomes. Our survey provides a broad perspective of pathologists' perceptions and experience regarding incorporation of histomolecular approaches to “personalize” therapy. Due to variable clinical adoption, there is a need for additional data using uniform study criteria. This will drive generation of best practice guidelines in this area for widespread and consistent c","PeriodicalId":13219,"journal":{"name":"Histopathology","volume":"85 5","pages":"748-759"},"PeriodicalIF":3.9,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141792354","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}
Loss of expression of tumour suppressor PAX2 and MMR deficiency (dMMR) has been frequently seen in endometrial endometrioid adenocarcinoma (EEC). However, the relationship between PAX2 expression and MMR status is unknown.
Methods and Results
We studied the PAX2 expression and examined its association with MMR status at the protein and genetic levels in 180 cases of EEC. Overall, total loss of PAX2 expression was found in about 70%, while retained PAX2 expression was seen in 30% of EEC. Among 125 cases with loss of PAX2, 68.8% were found in EECs with pMMR, while 31.2% were seen in those with dMMR. Among 55 cases of EECs with retained PAX2 expression, 92.7% were EECs with dMMR and 7.3% were those with pMMR (P < 0.001). While dMMR cases with MLH1 hypermethylation show almost equal retained or loss of PAX2 expression (52% versus 48%), dMMR with genetic alterations had significantly more retained PAX2 expression than loss of PAX2 (92.3% versus 7.7%), regardless of somatic or germline mutations. Loss of PAX2 was observed in 97.3% of dMMR with MLH1 hypermethylation compared to 2.7% of dMMR with genetic alterations (P < 0.001). Aggressive features such as higher tumour grades (FIGO 2–3) and advanced clinical stage (T2–T4) were significantly more frequently seen in dMMR with retained PAX2 expression, compared those to pMMR with loss of PAX2 expression.
Conclusion
Our study demonstrates a close correlation between retained PAX2 expression and dMMR in EEC. The molecular mechanism and clinical significance linking these two pathways in EEC remains to be unravelled.
{"title":"Retained PAX2 expression associated with DNA mismatch repair deficiency in endometrial endometrioid adenocarcinoma","authors":"Gloria X. Zhang, Bin Yang","doi":"10.1111/his.15281","DOIUrl":"10.1111/his.15281","url":null,"abstract":"<div>\u0000 \u0000 <section>\u0000 \u0000 <h3> Aims</h3>\u0000 \u0000 <p>Loss of expression of tumour suppressor PAX2 and MMR deficiency (dMMR) has been frequently seen in endometrial endometrioid adenocarcinoma (EEC). However, the relationship between PAX2 expression and MMR status is unknown.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods and Results</h3>\u0000 \u0000 <p>We studied the PAX2 expression and examined its association with MMR status at the protein and genetic levels in 180 cases of EEC. Overall, total loss of PAX2 expression was found in about 70%, while retained PAX2 expression was seen in 30% of EEC. Among 125 cases with loss of PAX2, 68.8% were found in EECs with pMMR, while 31.2% were seen in those with dMMR. Among 55 cases of EECs with retained PAX2 expression, 92.7% were EECs with dMMR and 7.3% were those with pMMR (<i>P</i> < 0.001). While dMMR cases with <i>MLH1</i> hypermethylation show almost equal retained or loss of PAX2 expression (52% versus 48%), dMMR with genetic alterations had significantly more retained PAX2 expression than loss of PAX2 (92.3% versus 7.7%), regardless of somatic or germline mutations. Loss of PAX2 was observed in 97.3% of dMMR with <i>MLH1</i> hypermethylation compared to 2.7% of dMMR with genetic alterations (<i>P</i> < 0.001). Aggressive features such as higher tumour grades (FIGO 2–3) and advanced clinical stage (T2–T4) were significantly more frequently seen in dMMR with retained PAX2 expression, compared those to pMMR with loss of PAX2 expression.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Our study demonstrates a close correlation between retained PAX2 expression and dMMR in EEC. The molecular mechanism and clinical significance linking these two pathways in EEC remains to be unravelled.</p>\u0000 </section>\u0000 </div>","PeriodicalId":13219,"journal":{"name":"Histopathology","volume":"85 5","pages":"794-803"},"PeriodicalIF":3.9,"publicationDate":"2024-07-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1111/his.15281","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141792356","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}
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