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Article ; Online: Unusual Patterns of HER2 Expression in Breast Cancer: Insights and Perspectives.

Grassini, Dora / Cascardi, Eliano / Sarotto, Ivana / Annaratone, Laura / Sapino, Anna / Berrino, Enrico / Marchiò, Caterina

Pathobiology : journal of immunopathology, molecular and cellular biology

2022 Volume 89, Issue 5, Page(s) 278–296

Abstract: The biomarker human epidermal growth factor receptor-2 (HER2) has represented the best example of successful targeted therapy in breast cancer patients. Based on the concept of "oncogene addiction," we have learnt how to identify patients likely ... ...

Abstract	The biomarker human epidermal growth factor receptor-2 (HER2) has represented the best example of successful targeted therapy in breast cancer patients. Based on the concept of "oncogene addiction," we have learnt how to identify patients likely benefitting from anti-HER2 agents. Since HER2 gene amplification leads to marked overexpression of the HER2 receptors on the cell membrane, immunohistochemistry with clinically validated antibodies and scoring system based on intensity and completeness of the membranous expression constitute the screening method to separate negative (score 0/1+) and positive (score 3+) carcinomas and to identify those tumours with complete yet only moderate HER2 expression (score 2+, equivocal carcinomas), which need to be investigated further in terms of gene status to confirm the presence of a loop of oncogene addiction. This process has demanded quality controls and led to recommendations by Scientific Societies, which pathologists routinely need to follow to guarantee reproducibility. In this review, we will span from the description of classical HER2 evaluation to the discussion of those scenarios in which HER2 expression is unusual and/or difficult to define. We will dissect HER2 heterogeneity, HER2 conversion from primary to relapsed/metastatic breast cancer, and we will introduce the new category of HER2-low breast carcinomas.
MeSH term(s)	Biomarkers, Tumor/genetics ; Breast Neoplasms/diagnosis ; Breast Neoplasms/genetics ; Breast Neoplasms/metabolism ; Carcinoma/genetics ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; Oncogene Addiction/genetics ; Receptor, ErbB-2/genetics ; Receptor, ErbB-2/metabolism ; Reproducibility of Results
Chemical Substances	Biomarkers, Tumor ; ERBB2 protein, human (EC 2.7.10.1) ; Receptor, ErbB-2 (EC 2.7.10.1)
Language	English
Publishing date	2022-05-02
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	1022703-9
ISSN	1423-0291 ; 1015-2008
ISSN (online)	1423-0291
ISSN	1015-2008
DOI	10.1159/000524227
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Article ; Online: Correction to: Real-world histopathological approach to malignancy of undefined primary origin (MUO) to diagnose cancers of unknown primary (CUPs).

Pisacane, Alberto / Cascardi, Eliano / Berrino, Enrico / Polidori, Alessio / Sarotto, Ivana / Casorzo, Laura / Panero, Mara / Boccaccio, Carla / Verginelli, Federica / Benvenuti, Silvia / Dellino, Miriam / Comoglio, Paolo / Montemurro, Filippo / Geuna, Elena / Marchiò, Caterina / Sapino, Anna

Virchows Archiv : an international journal of pathology

2022 Volume 482, Issue 3, Page(s) 477

Language	English
Publishing date	2022-12-17
Publishing country	Germany
Document type	Published Erratum
ZDB-ID	1184867-4
ISSN	1432-2307 ; 0945-6317
ISSN (online)	1432-2307
ISSN	0945-6317
DOI	10.1007/s00428-022-03475-5
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Article ; Online: Real-world histopathological approach to malignancy of undefined primary origin (MUO) to diagnose cancers of unknown primary (CUPs).

Virchows Archiv : an international journal of pathology

2022 Volume 482, Issue 3, Page(s) 463–475

Abstract: The aim of this study is to envisage a streamlined pathological workup to rule out CUPs in patients presenting with MUOs. Sixty-four MUOs were classified using standard histopathology. Clinical data, immunocytochemical markers, and results of molecular ... ...

Abstract	The aim of this study is to envisage a streamlined pathological workup to rule out CUPs in patients presenting with MUOs. Sixty-four MUOs were classified using standard histopathology. Clinical data, immunocytochemical markers, and results of molecular analysis were recorded. MUOs were histologically subdivided in clear-cut carcinomas (40 adenocarcinomas, 11 squamous, and 3 neuroendocrine carcinomas) and unclear-carcinoma features (5 undifferentiated and 5 sarcomatoid tumors). Cytohistology of 7/40 adenocarcinomas suggested an early metastatic cancer per se. In 33/40 adenocarcinomas, CK7/CK20 expression pattern, gender, and metastasis sites influenced tissue-specific marker selection. In 23/40 adenocarcinomas, a "putative-immunophenotype" of tissue of origin addressed clinical-diagnostic examinations, identifying 9 early metastatic cancers. Cell lineage markers were used to confirm squamous and neuroendocrine differentiation. Pan-cytokeratins were used to confirm the epithelial nature of poorly differentiated tumors, followed by tissue and cell lineage markers, which identified one melanoma. In total, 47/64 MUOs (73.4%) were confirmed CUP. Molecular analysis, feasible in 37/47 CUPs (78.7%), had no diagnostic impact. Twenty CUP patients, mainly with squamous carcinomas and adenocarcinomas with putative-gynecologic-immunophenotypes, presented with only lymph node metastases and had longer median time to progression and overall survival (< 0.001), compared with patients with other metastatic patterns. We propose a simplified histology-driven workup which could efficiently rule out CUPs and identify early metastatic cancer.
MeSH term(s)	Humans ; Female ; Neoplasms, Unknown Primary/diagnosis ; Neoplasms, Unknown Primary/pathology ; Immunohistochemistry ; Adenocarcinoma/metabolism ; Keratins/analysis ; Carcinoma, Squamous Cell/diagnosis ; Biomarkers, Tumor/analysis
Chemical Substances	Keratins (68238-35-7) ; Biomarkers, Tumor
Language	English
Publishing date	2022-11-08
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1184867-4
ISSN	1432-2307 ; 0945-6317
ISSN (online)	1432-2307
ISSN	0945-6317
DOI	10.1007/s00428-022-03435-z
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Article: Paclitaxel Restores Sensitivity to Chemotherapy in Preclinical Models of Multidrug-Resistant Intrahepatic Cholangiocarcinoma.

Massa, Annamaria / Peraldo-Neia, Caterina / Vita, Francesca / Varamo, Chiara / Basiricò, Marco / Raggi, Chiara / Bernabei, Paola / Erriquez, Jessica / Sarotto, Ivana / Leone, Francesco / Marchiò, Serena / Cavalloni, Giuliana / Aglietta, Massimo

Frontiers in oncology

2022 Volume 12, Page(s) 771418

Abstract: The treatment of unresectable cholangiocarcinoma (CCA) is limited by the development of resistance to conventional first-line chemotherapy based on gemcitabine (GEM). In addition, a prior treatment with GEM frequently induces cross-resistance to other ... ...

Abstract	The treatment of unresectable cholangiocarcinoma (CCA) is limited by the development of resistance to conventional first-line chemotherapy based on gemcitabine (GEM). In addition, a prior treatment with GEM frequently induces cross-resistance to other drugs employed in the second-line. Paclitaxel (PTX) is now emerging as an alternative option for the management of advanced/metastatic CCA. In the present work, we evaluate the antitumor activity of PTX in preclinical models of multidrug-resistant intrahepatic cholangiocarcinoma (iCCA).
Language	English
Publishing date	2022-02-17
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2649216-7
ISSN	2234-943X
ISSN	2234-943X
DOI	10.3389/fonc.2022.771418
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Article ; Online: The Multifaceted Nature of Tumor Microenvironment in Breast Carcinomas.

Annaratone, Laura / Cascardi, Eliano / Vissio, Elena / Sarotto, Ivana / Chmielik, Ewa / Sapino, Anna / Berrino, Enrico / Marchiò, Caterina

Pathobiology : journal of immunopathology, molecular and cellular biology

2020 Volume 87, Issue 2, Page(s) 125–142

Abstract: Heterogeneity in breast carcinomas can be appreciated at various levels, from morphology to molecular alterations, and there are well-known genotypic-phenotypic correlations. Clinical decision-making is strictly focused on the evaluation of tumor cells ... ...

Abstract	Heterogeneity in breast carcinomas can be appreciated at various levels, from morphology to molecular alterations, and there are well-known genotypic-phenotypic correlations. Clinical decision-making is strictly focused on the evaluation of tumor cells and is based on the assessment of hormone receptors and of the HER2 status, by means of a combination of immunohistochemical and in situ hybridization techniques. The tumor microenvironment (TME) also shows a multifaceted nature stemming from the different actors populating the intratumoral and the peritumoral stroma of breast carcinomas. Of note, we have now evidence that tumor-infiltrating lymphocytes (TILs) are clinically meaningful as their quantification in the intratumoral stroma strongly correlates with good prognosis, in particular in triple-negative and HER2-positive breast cancer patients. Nevertheless, TILs are just one of the many actors orchestrating the complexity of the TME, which is populated by immune and non-immune cells (cancer-associated fibroblasts, cancer-associated adipocytes), as well as non-cellular components such as chemical inflammation mediators. In this review article we will overview the main features of the distinct cell compartments by discussing (i) the potential impact the TME may have on the prognostic stratification of breast cancers and (ii) the possible predictive value of some markers in the context of immunotherapy in light of the recent results of phase III studies in advanced and early triple-negative breast cancer patients.
MeSH term(s)	Biomarkers, Tumor ; Breast Neoplasms/immunology ; Breast Neoplasms/physiopathology ; Breast Neoplasms/therapy ; Female ; Fibroblasts/pathology ; Humans ; Lymphocytes, Tumor-Infiltrating/pathology ; Prognosis ; Triple Negative Breast Neoplasms/pathology ; Tumor Microenvironment
Chemical Substances	Biomarkers, Tumor
Language	English
Publishing date	2020-04-23
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	1022703-9
ISSN	1423-0291 ; 1015-2008
ISSN (online)	1423-0291
ISSN	1015-2008
DOI	10.1159/000507055
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Article: Pursuit of Gene Fusions in Daily Practice: Evidence from Real-World Data in Wild-Type and Microsatellite Instable Patients.

Berrino, Enrico / Bragoni, Alberto / Annaratone, Laura / Fenocchio, Elisabetta / Carnevale-Schianca, Fabrizio / Garetto, Lucia / Aglietta, Massimo / Sarotto, Ivana / Casorzo, Laura / Venesio, Tiziana / Sapino, Anna / Marchiò, Caterina

Cancers

2021 Volume 13, Issue 13

Abstract: Agnostic biomarkers such as gene fusions allow to address cancer patients to targeted therapies; however, the low prevalence of these alterations across common malignancies poses challenges and needs a feasible and sensitive diagnostic process. RNA-based ...

Abstract	Agnostic biomarkers such as gene fusions allow to address cancer patients to targeted therapies; however, the low prevalence of these alterations across common malignancies poses challenges and needs a feasible and sensitive diagnostic process. RNA-based targeted next generation sequencing was performed on 125 samples of patients affected either by colorectal carcinoma, melanoma, or lung adenocarcinoma lacking genetic alterations in canonical driver genes, or by a colorectal carcinoma with microsatellite instability. Gene fusion rates were compared with in silico data from MSKCC datasets. For
Language	English
Publishing date	2021-07-05
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2527080-1
ISSN	2072-6694
ISSN	2072-6694
DOI	10.3390/cancers13133376
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Article ; Online: Reverse signaling by semaphorin 4C elicits SMAD1/5- and ID1/3-dependent invasive reprogramming in cancer cells.

Gurrapu, Sreeharsha / Franzolin, Giulia / Fard, Damon / Accardo, Massimo / Medico, Enzo / Sarotto, Ivana / Sapino, Anna / Isella, Claudio / Tamagnone, Luca

Science signaling

2019 Volume 12, Issue 595

Abstract: Semaphorins are a family of molecular signals that guide cell migration and are implicated in the regulation of cancer cells. In particular, transmembrane semaphorins are postulated to act as both ligands ("forward" mode) and signaling receptors (" ... ...

Abstract	Semaphorins are a family of molecular signals that guide cell migration and are implicated in the regulation of cancer cells. In particular, transmembrane semaphorins are postulated to act as both ligands ("forward" mode) and signaling receptors ("reverse" mode); however, reverse semaphorin signaling in cancer is relatively less understood. Here, we identified a previously unknown function of transmembrane semaphorin 4C (Sema4C), acting in reverse mode, to elicit nonconventional TGF-β/BMP receptor activation and selective SMAD1/5 phosphorylation. Sema4C coimmunoprecipitated with TGFBRII and BMPR1, supporting its role as modifier of this pathway. Sema4C reverse signaling led to the increased abundance of ID1/3 transcriptional factors and to extensive reprogramming of gene expression, which suppressed the typical features of the epithelial-mesenchymal transition in invasive carcinoma cells. This phenotype was nevertheless coupled with burgeoning metastatic behavior in vivo, consistent with evidence that Sema4C expression correlates with metastatic progression in human breast cancers. Thus, Sema4C reverse signaling promoted SMAD1/5- and ID1/3-dependent gene expression reprogramming and phenotypic plasticity in invasive cancer cells.
MeSH term(s)	Animals ; COS Cells ; Chlorocebus aethiops ; Humans ; Inhibitor of Differentiation Protein 1/genetics ; Inhibitor of Differentiation Protein 1/metabolism ; Inhibitor of Differentiation Proteins/genetics ; Inhibitor of Differentiation Proteins/metabolism ; Neoplasm Invasiveness ; Neoplasm Proteins/genetics ; Neoplasm Proteins/metabolism ; Neoplasms/genetics ; Neoplasms/metabolism ; Neoplasms/pathology ; PC-3 Cells ; Semaphorins/genetics ; Semaphorins/metabolism ; Signal Transduction ; Smad1 Protein/genetics ; Smad1 Protein/metabolism ; Smad3 Protein/genetics ; Smad3 Protein/metabolism
Chemical Substances	ID1 protein, human ; Inhibitor of Differentiation Protein 1 ; Inhibitor of Differentiation Proteins ; Neoplasm Proteins ; SMAD1 protein, human ; SMAD3 protein, human ; Sema4c protein, human ; Semaphorins ; Smad1 Protein ; Smad3 Protein ; ID3 protein, human (147785-34-0)
Language	English
Publishing date	2019-08-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2417226-1
ISSN	1937-9145 ; 1945-0877
ISSN (online)	1937-9145
ISSN	1945-0877
DOI	10.1126/scisignal.aav2041
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Article ; Online: Integrative genomic and transcriptomic analyses illuminate the ontology of HER2-low breast carcinomas.

Genome medicine

2022 Volume 14, Issue 1, Page(s) 98

Abstract: Background: The "HER2-low" nomenclature identifies breast carcinomas (BCs) displaying a HER2 score of 1+/2+ in immunohistochemistry and lacking ERBB2 amplification. Whether HER2-low BCs (HLBCs) constitute a distinct entity is debated.: Methods: We ... ...

Abstract	Background: The "HER2-low" nomenclature identifies breast carcinomas (BCs) displaying a HER2 score of 1+/2+ in immunohistochemistry and lacking ERBB2 amplification. Whether HER2-low BCs (HLBCs) constitute a distinct entity is debated. Methods: We performed DNA and RNA high-throughput analysis on 99 HLBC samples (n = 34 cases with HER2 score 1+/HLBC-1, n = 15 cases with HER2 score 2+ and ERBB2 not amplified/HLBC-2N, and n = 50 cases with score 2+ and ERBB2 copy number in the equivocal range/HLBC-2E). We compared the mutation rates with data from 1317 samples in the Memorial Sloan-Kettering Cancer Center (MSKCC) BC cohort and gene expression data with those from an internal cohort of HER2-negative and HER2-positive BCs. Results: The most represented mutations affected PIK3CA (31/99, 31%), GATA3 (18/99, 18%), TP53 (17/99, 17%), and ERBB2 (8/99, 8%, private to HLBC-2E). Tumor mutational burden was significantly higher in HLBC-1 compared to HLBC-2E/N (P = 0.04). Comparison of mutation spectra revealed that HLBCs were different from both HER2-negative and HER2-positive BCs, with HLBC-1 resembling more HER2-negative tumors and HLBC-2 mutationally related to HER2-addicted tumors. Potentially actionable alterations (annotated by using OncoKB/ESCAT classes) affected 52 patients. Intra-group gene expression revealed overlapping features between HLBC-1 and control HER2-negative BCs, whereas the HLBC-2E tumors showed the highest diversity overall. The RNA-based class discovery analysis unveiled four subsets of tumors with (i) lymphocyte activation, (ii) unique enrichment in HER2-related features, (iii) stromal remodeling alterations, and (iv) actionability of PIK3CA mutations (LAURA classification). Conclusions: HLBCs harbor distinct genomic features when compared with HER2-positive and HER2-negative BCs; however, differences across IHC classes were also unveiled thus dissecting the full picture of heterogeneity across HER2-low disease. The HLBC-2E category harbors most distinctive features, whereas HLBC-1 seems superimposable to HER2-negative disease. Further studies are needed to ascertain whether the four genomic-driver classes of the LAURA classification hold prognostic and/or predictive implications.
MeSH term(s)	Biomarkers, Tumor/genetics ; Breast Neoplasms/pathology ; Female ; Gene Expression Profiling ; Genomics ; Humans ; Mutation ; RNA ; Transcriptome
Chemical Substances	Biomarkers, Tumor ; RNA (63231-63-0)
Language	English
Publishing date	2022-08-29
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2484394-5
ISSN	1756-994X ; 1756-994X
ISSN (online)	1756-994X
ISSN	1756-994X
DOI	10.1186/s13073-022-01104-z
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Article ; Online: Aurora kinase A gene copy number is associated with the malignant transformation of colorectal adenomas but not with the serrated neoplasia progression.

Casorzo, Laura / Dell'Aglio, Carmine / Sarotto, Ivana / Risio, Mauro

Human pathology

2015 Volume 46, Issue 3, Page(s) 411–418

Abstract: A crucial role for Aurora Kinase A (AURKA) gene has been demonstrated in the advanced steps of colorectal tumor progression. Little is known, however, about its role in the early phases of the adenoma-carcinoma sequence. Moreover, no data are currently ... ...

Abstract	A crucial role for Aurora Kinase A (AURKA) gene has been demonstrated in the advanced steps of colorectal tumor progression. Little is known, however, about its role in the early phases of the adenoma-carcinoma sequence. Moreover, no data are currently available concerning AURKA involvement in the serrated tumorigenesis. Fluorescence in situ hybridization analysis and immunohistochemistry were used to assess gene copy number and protein expression in 40 colorectal adenomas, 20 cancerized adenomas, and 20 serrated polyps. An increased copy number was found either in adenomatous tissue or in early cancer in the vast majority of cancerized adenomas, but only in 5% of adenomas (P < .001). Protein expression strictly paralleled fluorescence in situ hybridization results. No changes in the gene copy number were observed in serrated polyps, regardless of their histotype and the presence of dysplasia, even if high percentages of immunostained cells were detected in all the subgroups. AURKA gene is associated with progressive colorectal adenomas but is uninvolved in the development of nonprogressive adenomas. The diploid status of the gene is maintained along the progression of serrated neoplasia. AURKA protein expression in serrated polyps is uncoupled from gene status and is likely to reflect apoptotic dysregulation.
MeSH term(s)	Adenoma/enzymology ; Adenoma/genetics ; Adenoma/pathology ; Aurora Kinase A/genetics ; Cell Transformation, Neoplastic/genetics ; Cell Transformation, Neoplastic/metabolism ; Cell Transformation, Neoplastic/pathology ; Colonic Polyps/enzymology ; Colonic Polyps/genetics ; Colonic Polyps/pathology ; Colorectal Neoplasms/enzymology ; Colorectal Neoplasms/genetics ; Colorectal Neoplasms/pathology ; Disease Progression ; Gene Dosage/genetics ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Neoplasm Grading ; Neoplasm Invasiveness/genetics ; Neoplasm Invasiveness/pathology
Chemical Substances	Aurora Kinase A (EC 2.7.11.1)
Language	English
Publishing date	2015-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	207657-3
ISSN	1532-8392 ; 0046-8177
ISSN (online)	1532-8392
ISSN	0046-8177
DOI	10.1016/j.humpath.2014.11.016
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Article: Cold Formalin Fixation Guarantees DNA Integrity in Formalin Fixed Paraffin Embedded Tissues: Premises for a Better Quality of Diagnostic and Experimental Pathology With a Specific Impact on Breast Cancer.

Berrino, Enrico / Annaratone, Laura / Miglio, Umberto / Maldi, Elena / Piccinelli, Chiara / Peano, Erica / Balmativola, Davide / Cassoni, Paola / Pisacane, Alberto / Sarotto, Ivana / Venesio, Tiziana / Sapino, Anna / Marchiò, Caterina

Frontiers in oncology

2020 Volume 10, Page(s) 173

Abstract: Formalin fixation and paraffin embedding (FFPE) represent the standard method to preserve tissue specimens for diagnostic pathology, however formalin fixation induces severe fragmentation of nucleic acids. We investigated whether formalin fixation at 4°C ...

Abstract	Formalin fixation and paraffin embedding (FFPE) represent the standard method to preserve tissue specimens for diagnostic pathology, however formalin fixation induces severe fragmentation of nucleic acids. We investigated whether formalin fixation at 4°C could preserve DNA integrity in FFPE specimens. Paired samples from 38 specimens were formalin fixed at room temperature (
Language	English
Publishing date	2020-02-19
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2649216-7
ISSN	2234-943X
ISSN	2234-943X
DOI	10.3389/fonc.2020.00173
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