Journal of Personalized Medicine (Apr 2022)

Validation of a Novel Three-Dimensional (<i>3D Fusion</i>) Gross Sampling Protocol for Clear Cell Renal Cell Carcinoma to Overcome Intratumoral Heterogeneity: The Meet-Uro 18 Study

  • 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

DOI
https://doi.org/10.3390/jpm12050727
Journal volume & issue
Vol. 12, no. 5
p. 727

Abstract

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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.

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