We aimed to overcome intratumoral heterogeneity in clear cell renal cell carcinoma (clearRCC). One hundred cases of clearRCC were sampled. First, usual standard sampling was applied (1 block/cm of tumor); second, the whole tumor was sampled, and 0. 6 mm cores were taken from each block to construct a tissue microarray; third, the residual tissue, mapped by taking pieces 0.5 × 0.5 cm, reconstructed the entire tumor mass. Precisely, six randomly derived pieces of tissues were placed in each cassette, with the number of cassettes being based on the diameter of the tumor (called multisite 3D fusion). Angiogenic and immune markers were tested. Routine 5231 tissue blocks were obtained. Multisite 3D fusion sections showed pattern A, homogeneous high vascular density (10%), pattern B, homogeneous low vascular density (8%) and pattern C, heterogeneous angiogenic signatures (82%). PD-L1 expression was seen as diffuse (7%), low (33%) and absent (60%). Tumor-infiltrating CD8 scored high in 25% (pattern hot), low in 65% (pattern weak) and zero in 10% of cases (pattern desert). Grading was upgraded in 26% of cases (G3-G4), necrosis and sarcomatoid/rhabdoid characters were observed in, respectively, 11 and 7% of cases after 3D fusion (p = 0.03). CD8 and PD-L1 immune expressions were higher in the undifferentiated G4/rhabdoid/sarcomatoid clearRCC subtypes (p = 0.03). Again, 22% of cases were set to intermediate to high risk of clinical recurrence due to new morphological findings of all aggressive G4, sarcomatoid/rhabdoid features by using 3D fusion compared to standard methods (p = 0.04). In conclusion, we propose an easy-to-apply multisite 3D fusion sampling that negates bias due to tumor heterogeneity.
Journal of personalized medicine. 2022 Apr 30*** epublish ***
Matteo Brunelli, Guido Martignoni, Giorgio Malpeli, Alessandro Volpe, Luca Cima, Maria Rosaria Raspollini, Mattia Barbareschi, Alessandro Tafuri, Giulia Masi, Luisa Barzon, Serena Ammendola, Manuela Villanova, Maria Angela Cerruto, Michele Milella, Sebastiano Buti, Melissa Bersanelli, Giuseppe Fornarini, Sara Elena Rebuzzi, Valerio Gaetano Vellone, Gabriele Gaggero, Giuseppe Procopio, Elena Verzoni, Sergio Bracarda, Martina Fanelli, Roberto Sabbatini, Rodolfo Passalacqua, Bruno Perrucci, Maria Olga Giganti, Maddalena Donini, Stefano Panni, Marcello Tucci, Veronica Prati, Cinzia Ortega, Anna Caliò, Albino Eccher, Filippo Alongi, Giovanni Pappagallo, Roberto Iacovelli, Alessandra Mosca, Paolo Umari, Ilaria Montagnani, Stefano Gobbo, Francesco Atzori, Enrico Munari, Marco Maruzzo, Umberto Basso, Francesco Pierconti, Carlo Patriarca, Piergiuseppe Colombo, Alberto Lapini, Giario Conti, Roberto Salvioni, Enrico Bollito, Andrea Cossarizza, Francesco Massari, Mimma Rizzo, Renato Franco, Federica Zito-Marino, Yoseba Aberasturi Plata, Francesca Galuppini, Marta Sbaraglia, Matteo Fassan, Angelo Paolo Dei Tos, Maurizio Colecchia, Holger Moch, Maurizio Scaltriti, Camillo Porta, Brett Delahunt, Gianluca Giannarini, Roberto Bortolus, Pasquale Rescigno, Giuseppe Luigi Banna, Alessio Signori, Miguel Angel Llaja Obispo, Roberto Perris, Alessandro Antonelli
Pathology Unit, Department of Diagnostics and Public Health, University and Hospital Trust of Verona, 37134 Verona, Italy., Department of Surgery, Dentistry, Paediatrics and Gynaecology, University of Verona, 37134 Verona, Italy., Division of Urology, Maggiore della Carità Hospital, 28100 Novara, Italy., Pathology Unit, Department of Clinical Services, Santa Chiara Hospital, 38100 Trento, Italy., Histopathology and Molecular Diagnostics, University Hospital Careggi, 50100 Firenze, Italy., Division of Urology, Department of Surgery, Dentistry, Paediatrics and Gynaecology, University and Hospital Trust of Verona, 37134 Verona, Italy., Department of Molecular Medicine, University of Padua, 35100 Padova, Italy., Division of Oncology, University and Hospital Trust of Verona, 37134 Verona, Italy., Division of Oncology, University and Hospital Trust of Parma, 43100 Parma, Italy., Division of Oncology, San Martino Hospital, 16100 Genova, Italy., Pathology Unit, San Martino Hospital, 16100 Genova, Italy., Division of Oncology, IRCCS Foundation, Istituto Nazionale dei Tumori di Milano, 20133 Milano, Italy., Division of Oncology, Santa Maria Hospital, 05100 Terni, Italy., Division of Oncology, Department of Oncology, Hematology & Respiratory Diseases, University of Modena & Reggio Emilia, 41121 Modena, Italy., Division of Oncology, Hospital Trust of Cremona, 26100 Cremona, Italy., Division of Oncology, Cardinal Massaia Hospital, 14100 Asti, Italy., Division of Oncology, Institute for Cancer Research and Treatment, Asl Cn2 Alba-Brà, 12042 Cuneo, Italy., Advanced Radiation Oncology Department, IRCCS Ospedale Sacro Cuore Don Calabria, Negrar, 37024 Verona, Italy., Indipendent Researcher, Clinical Epidemiologist, Silea, 31057 Treviso, Italy., Policlinico Universitario Fondazione Andrea Gemelli, 00168 Roma, Italy., Oncology, Candiolo Cancer Institute, IRCCS-FPO, 10060 Torino, Italy., Department of Translation Medicine, Pathology Unit, University of Ferrara, 44100 Ferrara, Italy., Oncology Unit, University and Hospital Trust of Cagliari, 09100 Cagliari, Italy., Pathology Unit, Department of Molecular and Translational Medicine, University of Brescia, 25100 Brescia, Italy., Oncology Unit, Oncology Department, Istituto Oncologico Veneto (IOV), IRCCS, 35128 Padua, Italy., Division of Pathology, Policlinico Universitario Fondazione Agostino Gemelli, 00168 Roma, Italy., Pathology Unit, Department of Clinical Services, Sant'Anna Hospital, 22100 Como, Italy., Department of Pathology, Department of Biomedical Sciences, Humanitas University, IRCCS Humanitas Clinical and Research Hospital, Pieve Emanuele, 20089 Milan, Italy., Division of Urology, University Hospital Careggi, 50100 Firenze, Italy., Division of Urology, Sant'Anna Hospital, 22100 Como, Italy., Department of Urology, Fondazione IRCCS Istituto Nazionale dei Tumori di Milano, 20133 Milano, Italy., Department of Pathology, San Luigi Gonzaga Hospital, University of Turin, 10122 Torino, Italy., Department of Medical and Surgical Sciences for Children and Adults, University of Modena & Reggio Emilia, 41121 Modena, Italy., Medical Oncology, University and Hospital Trust of Bologna, 40138 Bologna, Italy., Division of Translational Oncology, IRCCS Istituti Clinici Scientifici 'Maugeri', 27100 Pavia, Italy., Pathology Unit, Department of Mental and Physical Health and Preventive Medicine, University of Campania 'Luigi Vanvitelli', 81100 Caserta, Italy., Department of Pathology, Cruces University Hospital, University of the Basque Country (UPV/EHU), 48900 Barakaldo, Spain., Department of Pathology, University and Hospital Trust of Padua, 35128 Padova, Italy., Department of Pathology, Uropathology Unit, Vita-Salute San Raffaele University, IRCCS San Raffaele, 20132 Milano, Italy., Department of Pathology and Molecular Pathology, University Hospital Zurich, 8006 Zurich, Switzerland., AstraZeneca, New York, NY 10005, USA., Department of Biomedical Sciences and Human Oncology, University of Bari 'A. Moro', 70121 Bari, Italy., Department of Pathology and Molecular Medicine, Wellington School of Medicine and Health Sciences, University of Otago, 3743 Wellington, New Zealand., Urology Unit, "Santa Maria della Misericordia", University of Udine, 33100 Udine, Italy., Department of Radiotherapy, CRO, 33081 Aviano, Italy., Candiolo Cancer Institute, FPO-IRCCS, Candiolo, 10060 Turin, Italy., Department of Health Sciences (DISSAL), Section of Biostatistics, University of Genova, 16100 Genova, Italy., Medical Oncology Unit 1, IRCCS Ospedale Policlinico San Martino, 16100 Genova, Italy., Department of Biosciences, COMT-Centre for Molecular and Translational Oncology, University of Parma, 43124 Parma, Italy.