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Hand2 Immunohistochemistry in the Diagnosis of Paragangliomas and Other Neuroendocrine Neoplasms

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Abstract

Hand2 is a core transcription factor responsible for chromaffin cell differentiation. However, its potential utility in surgical pathology has not been studied. Thus, we aimed to investigate its expression in paragangliomas, other neuroendocrine neoplasms (NENs), and additional non-neuroendocrine tumors. We calibrated Hand2 immunohistochemistry on adrenal medulla cells and analyzed H-scores in 46 paragangliomas (PGs), 9 metastatic PGs, 21 cauda equina neuroendocrine tumors (CENETs), 48 neuroendocrine carcinomas (NECs), 8 olfactory neuroblastomas (ONBs), 110 well-differentiated NETs (WDNETs), 10 adrenal cortical carcinomas, 29 adrenal cortical adenomas, 8 melanomas, 41 different carcinomas, and 10 gastrointestinal stromal tumors (GISTs). Both tissue microarrays (TMAs) and whole sections (WSs) were studied. In 171 NENs, previously published data on Phox2B and GATA3 were correlated with Hand2. Hand2 was positive in 98.1% (54/55) PGs, but only rarely in WDNETs (9.6%, 10/104), CENETs (9.5%, 2/21), NECs (4.2%, 2/48), or ONBs (12.5%, 1/8). Any Hand2 positivity was 98.1% sensitive and 91.7% specific for the diagnosis of PG. The Hand2 H-score was significantly higher in primary PGs compared to Hand2-positive WDNETs (median 166.3 vs. 7.5; p < 0.0001). Metastatic PGs were positive in 88.9% (8/9). No Hand2 positivity was observed in any adrenal cortical neoplasm or other non-neuroendocrine tumors, with exception of 8/10 GISTs. Parasympathetic PGs showed a higher Hand2 H-score compared to sympathetic PGs (median H-scores 280 vs. 104, p < 0.0001). Hand2 positivity in NENs serves as a reliable marker of primary and metastatic PG, since other NENs only rarely exhibit limited Hand2 positivity.

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Data Availability

The immunohistochemical results of individual tumors, together with basic clinical data, are available as Supplementary material. Additional clinical data and histological images (in the form of “*.svs” formatted image files) used and analyzed during the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

We would like to thank Dr. Boris Rychly (Unilabs sro., Bratislava, SK), Dr. Marketa Trnkova (AeskuLab k.s., Prague, CR), Dr. Ludmila Michnova (Military University Hospital Prague, Prague, CR), Maria Wozniakova (University Hospital Ostrava and Faculty of Medicine, University of Ostrava), Dr. Lukas Krbal, and Dr. Hana Faistova (University Hospital Hradec Kralove) for their participation in constructing the dataset of the NENs. We would like to thank Mrs. Svetlana Kopecka, Mrs. Petra Satori, Mrs. Jana Potocna, and Mr. Michal Valasek for their assistance with immunohistochemistry procedures.

Funding

This study was supported by the project BBMRI-CZ LM2023033, Charles University Cooperatio Program, research area DIAG and METD and Project of Czech Ministry of Defense MO 1012. The work was supported by the European Regional Development Fund-Project BBMRI-CZ Biobank network—a versatile platform for the research of the etiopathogenesis of diseases, No: EF16_013/0001674. Supported by Ministry of Health, Czech Republic—conceptual development of research organization 00064165, General University Hospital in Prague and by Ministry of Health of the Czech Republic, grant NU23-01-00323 and RVO VFN64165, the Czech Science Foundation grant 22-07091S. Funding sources were not involved in study design, collection, analysis, or interpretation, in the writing of the report, or in the decision to submit the article for publication.

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Author notes

  1. Jiri Soukup and Monika Manethova contributed equally to the work.

Authors and Affiliations

  1. Department of Pathology, Military University Hospital Prague, U Vojenske Nemocnice 1200, Praha 6, 169 02, Prague, Czech Republic

    Jiri Soukup & Marie Novakova

  2. The Fingerland Department of Pathology, Charles University, Faculty of Medicine in Hradec Králové and University Hospital Hradec Králové, Sokolska 582, 500 05, Hradec Kralove, Czech Republic

    Jiri Soukup, Monika Manethova, Vaclav Stejskal, Helena Hornychova & Ales Ryska

  3. Department of Pathology, Charles University, First Faculty of Medicine and General University Hospital in Prague, Studnickova, 2039, 128 00, Nové Mesto, Prague, Czech Republic

    Jiri Soukup & Jaroslava Duskova

  4. Department of Pathology, The Regional Hospital Pardubice, Pardubice, Czech Republic

    Maria Hacova

  5. 1st Department of Pathology, St. Anne’s University Hospital and Faculty of Medicine , Masaryk University, Pekarská 664/53, 602 00, Brno-stred, Brno, Czech Republic

    Iva Staniczkova-Zambo

  6. Centre for Hypertension, 3rd Department of Medicine, Charles University, First Faculty of Medicine and General University Hospital in Prague, Prague, Czech Republic

    Tomas Zelinka

  7. Department of Internal Medicine, First Faculty of Medicine, Charles University and Military University Hospital, Prague, Czech Republic

    Mikulas Kosak

  8. Department of Neurosurgery, Charles University, Faculty of Medicine in Hradec Králové and University Hospital Hradec Králové, Sokolska 582, 500 05, Hradec Kralove, Czech Republic

    Tomas Cesak

  9. Department of Neurosurgery and Neurooncology, 1st Medical Faculty, Charles University, Military University Hospital Prague, U Vojenske Nemocnice 1200, Praha 6, 169 02, Prague, Czech Republic

    David Netuka

  10. 4th Department of Internal Medicine, Charles University, Faculty of Medicine in Hradec Králové and University Hospital Hradec Králové, Sokolska 582, 500 05, Hradec Kralove, Czech Republic

    Filip Gabalec

Authors
  1. Jiri Soukup
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Contributions

JS, FG, and AR conceived and designed the study; JS, JD, ISZ, MM, MN, and MH procured and reviewed the cases included in the study; TC and DN procured clinical data from tumors and histological material from CENET and helped with study design; MK, TZ, and FG procured clinical data; HH implemented, validated, and performed immunohistochemical procedures; MM, JS, FG, and VS analyzed the results of the study; JS and MM performed statistical analysis. All authors contributed to the writing and review of the manuscript.

Corresponding author

Correspondence to Jiri Soukup.

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Ethics Approval

The study was carried out in accordance with the Declaration of Helsinki and with the approval of the Ethics committee of the University Hospital Hradec Kralove (reference no. 202101P06 and no. 202109P01).

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The authors declare no competing interests.

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Soukup, J., Manethova, M., Stejskal, V. et al. Hand2 Immunohistochemistry in the Diagnosis of Paragangliomas and Other Neuroendocrine Neoplasms. Endocr Pathol 35, 14–24 (2024). https://doi.org/10.1007/s12022-024-09803-6

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