Original articles

Vol. 117: Issue 5 - October 2025

Harmonization trial of FGFR1-3 testing strategies in cholangiocarcinoma patients: an Italian multicenter experience

Authors

Francesco Pepe , Gianluca Russo , Claudia Scimone , Lucia Palumbo , Stefania Tommasi , Rosamaria Pinto , Dario De Biase , Thais Maloberti , Adele Busico , Alessandra Santoro , Domenico Salemi , Elisa Melucci , Domenico Cozzolino , Luisa Toffolatti , Silvia Bessi , Claudia Sarracino , Ilaria Tomaiuolo , Angelo Minucci , Giuseppina Roscigno , Francesco Esposito , Pierlorenzo Pallante , Sara Lonardi , Giancarlo Pruneri , Giancarlo Troncone , Matteo Fassan , Umberto Malapelle

Key words: cholangiocarcinoma, molecular biomarker, FGFR
DOI: 10.32074/1591-951X-1317
Publication Date: 2025-12-16

Abstract

Aims. Molecular analysis of FGFR2 aberrant transcripts became crucial for clinical stratification of intrahepatic cholangiocarcinoma (iCCA) patients. Several strategies, including fluorescent in situ hybridization (FISH) and next generation sequencing (NGS), are commonly used to investigate FGFR aberrations. Here, we evaluated the technical performance of clinically implemented diagnostic strategies in 8 referral Italian institutions on artificial reference formalin-fixed paraffin-embedded (FFPE) samples.
Methods. Each participating institution was requested to apply its own diagnostic testing strategy on 8 sections obtained from artificial reference specimens built to harbor FGFR3(17)-TACC3(11) rearrangement and unbalanced FGFR2. A second-round slide set hosting FGFR2(17)-BICC1(3) aberrant transcript was shared to detect clinically relevant FGFR2 fusion. Artificial reference sample was previously validated by the University of Naples Federico II before arranging the shipment. Technical procedures (e.g. extraction methods, testing platforms and assays) were recorded.
Results. Overall, cell resuspension yielded higher amounts of DNA and RNA (SNU16 61.5 ng/µl, 38100.0 pg/µl; RT112 118.0/µl, 2140.0 pg/µl, respectively) in comparison with SNU16+ RT112 mixing cell block (0.7 ng/µl DNA and 412.0 pg/µl RNA). Moreover, FFPE samples showed a higher fragmentation index (DIN 1.2 and RIN not calculated) compared with cell line resuspension (DIN 2.2 and 9.5 for SNU16 and RT112; RIN 3.9 and 6.8 for SNU16 and RT112). All participating institutions identified FGFR2(17)-BICC1(3) and FGFR3(17)-TACC3(11) aberrant transcripts. Moreover, ID#2, ID#4, ID#7 institutions also detected FGFR2(3)-CD44(1) rearrangement on RNA, whereas institutions ID#1, ID#2, ID#3, ID#5, ID#6, ID#8 identified FGFR2 CNVs on DNA.
Conclusions. NGS represents the most suitable approach in molecular profiling of FGFR aberrant transcripts. Rings trial based on artificial reference samples play a pivotal role in optimizing routine diagnostic procedures filling the gap in clinical stratification of iCCA patients.

Downloads

PDF
Supp. File
Authors

Francesco Pepe - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Gianluca Russo - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Claudia Scimone - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Lucia Palumbo - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Stefania Tommasi - Molecular Diagnostics and Pharmacogenetics Unit, IRCCS-Istituto Tumori "Giovanni Paolo II", Bari, Italy

Rosamaria Pinto - Molecular Diagnostics and Pharmacogenetics Unit, IRCCS-Istituto Tumori "Giovanni Paolo II", Bari, Italy

Dario De Biase - Solid Tumor Molecular Pathology Laboratory, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy. Department of Pharmacy and Biotechnology (FaBit), University of Bologna, Bologna, Italy.

Thais Maloberti - Solid Tumor Molecular Pathology Laboratory, IRCCS Azienda Ospedaliero-Universitaria di Bologna, Bologna, Italy.

Adele Busico - Department of Diagnostic Innovation, Pathology Unit 2, Fondazione IRCCS Istituto Nazionale Dei Tumori, Milan, Italy.

Alessandra Santoro - UOSD Laboratory of Oncohematology, Cellular Manipulation and Cytogenetics, Department of Genetic, Oncohematology a Rare Disease, AOR "Villa Sofia-Cervello", Palermo, Italy.

Domenico Salemi - UOSD Laboratory of Oncohematology, Cellular Manipulation and Cytogenetics, Department of Genetic, Oncohematology a Rare Disease, AOR “Villa Sofia-Cervello”, Palermo, Italy

Elisa Melucci - Pathologic Anatomy and Histology Cytodiagnostics and Advanced Molecular Diagnostics, IRCCS Istituto Nazionale Tumori Regina Elena Roma.

Domenico Cozzolino - Department of Public Health, Federico II University of Naples, Naples, Italy

Luisa Toffolatti - Ca' Foncello General Hospital, Surgical Pathology Unit, ULSS2 Marca Trevigiana, Treviso (TV)

Silvia Bessi - Departmental Structure of Oncological Molecular Pathology, Oncological Department Azienda USL Toscana Centro, S. Stefano Hospital, Prato, Italy.

Claudia Sarracino - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Ilaria Tomaiuolo - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Angelo Minucci - Departmental Unit of Molecular and Genomic Diagnostics, Fondazione Policlinico Universitario Agostino Gemelli IRCCS, Rome, Italy.

Giuseppina Roscigno - Department of Biology, Federico II University of Naples, Naples, Italy.

Francesco Esposito - Institute of Endotypes in Oncology, Metabolism and Immunology (IEOMI) "G. Salvatore", National Research Council (CNR), Naples, Italy. Department of Molecular Medicine and Medical Biotechnology (DMMBM), University of Naples "Federico II", Naples, Italy.

Pierlorenzo Pallante - Institute of Endotypes in Oncology, Metabolism and Immunology (IEOMI) "G. Salvatore", National Research Council (CNR), Naples, Italy. Department of Molecular Medicine and Medical Biotechnology (DMMBM), University of Naples "Federico II", Naples, Italy.

Sara Lonardi - Oncology Unit 1, Department of Oncology Veneto Institute of Oncology IOV-IRCCS, Padova, Italy

Giancarlo Pruneri - Department of Diagnostic Innovation, Pathology Unit 2, Fondazione IRCCS Istituto Nazionale Dei Tumori, Milan, Italy. Department of Oncology and Haemato-Oncology, University of Milan, Milan, Italy.

Giancarlo Troncone - Department of Public Health, Federico II University of Naples, Via S. Pansini, 5, 80131 Naples, Italy.

Matteo Fassan - Department of Medicine - DIMED, University of Padua, Padua, Italy. Veneto Institute of Oncology IOV - IRCCS, Padua, Italy.

Umberto Malapelle - Università degli Studi di Napoli “Federico II”,

License

Creative Commons License

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Copyright

Copyright (c) 2025 Società Italiana di Anatomia Patologica e Citopatologia Diagnostica, Divisione Italiana della International Academy of Pathology

How to Cite
Pepe, F., Russo, G., Scimone, C., Palumbo, L., Tommasi, S., Pinto, R., De Biase, D., Maloberti, T., Busico, A., Santoro, A., Salemi, D., Melucci, E., Cozzolino, D., Toffolatti, L., Bessi, S., Sarracino, C., Tomaiuolo, I., Minucci, A., Roscigno, G., Esposito, F., Pallante, P., Lonardi, S., Pruneri, G., Troncone, G., Fassan, M., & Malapelle, U. (2025). Harmonization trial of FGFR1-3 testing strategies in cholangiocarcinoma patients: an Italian multicenter experience. Pathologica - Journal of the Italian Society of Anatomic Pathology and Diagnostic Cytopathology, 117(5). https://doi.org/10.32074/1591-951X-1317
  • Abstract viewed - 8 times
  • PDF downloaded - 0 times
  • Supp. File downloaded - 0 times