Intra-tumour heterogeneity of diffuse large B-cell lymphoma involves the induction of diversified stroma-tumour interfaces
Sabina Sangaletti,
Fabio Iannelli,
Federica Zanardi,
Valeria Cancila,
Paola Portararo,
Laura Botti,
Davide Vacca,
Claudia Chiodoni,
Arianna Di Napoli,
Cesare Valenti,
Celeste Rizzello,
Maria Carmela Vegliante,
Federica Pisati,
Alessandro Gulino,
Maurilio Ponzoni,
Mario Paolo Colombo,
Claudio Tripodo
Affiliations
Sabina Sangaletti
Molecular Immunology Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy
Fabio Iannelli
Bioinformatics Core Unit, IFOM, FIRC Institute of Molecular Oncology, Milan, Italy
Federica Zanardi
Bioinformatics Core Unit, IFOM, FIRC Institute of Molecular Oncology, Milan, Italy
Valeria Cancila
Tumor Immunology Unit, University of Palermo, Palermo, Italy
Paola Portararo
Molecular Immunology Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy
Laura Botti
Molecular Immunology Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy
Davide Vacca
Tumor Immunology Unit, University of Palermo, Palermo, Italy
Claudia Chiodoni
Molecular Immunology Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy
Arianna Di Napoli
Pathology Unit, Sapienza University of Rome, Sant'Andrea Hospital, Rome, Italy
Cesare Valenti
Department of Mathematics and Informatics, University of Palermo, Palermo, Italy
Celeste Rizzello
Molecular Immunology Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy
Maria Carmela Vegliante
Hematology and Cell Therapy Unit, IRCCS-Istituto Tumori 'Giovanni Paolo II', Bari, Italy
Federica Pisati
Tumor and Microenvironment Histopathology Unit, IFOM, FIRC Institute of Molecular Oncology, Milan, Italy
Alessandro Gulino
Tumor Immunology Unit, University of Palermo, Palermo, Italy
Maurilio Ponzoni
Pathology Unit, IRCCS San Raffaele Scientific Institute, Vita-Salute San Raffaele University Milan, Milan, Italy
Mario Paolo Colombo
Molecular Immunology Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milan, Italy; Corresponding author.
Claudio Tripodo
Tumor Immunology Unit, University of Palermo, Palermo, Italy; Tumor and Microenvironment Histopathology Unit, IFOM, FIRC Institute of Molecular Oncology, Milan, Italy; Corresponding author at: Tumor Immunology Unit, University of Palermo, Palermo, Italy.
Background: Intra-tumour heterogeneity in lymphoid malignancies encompasses selection of genetic events and epigenetic regulation of transcriptional programs. Clonal-related neoplastic cell populations are unsteadily subjected to immune editing and metabolic adaptations within different tissue microenvironments. How tissue-specific mesenchymal cells impact on the diversification of aggressive lymphoma clones is still unknown. Methods: Combining in situ quantitative immunophenotypical analyses and RNA sequencing we investigated the intra-tumour heterogeneity and the specific mesenchymal modifications that are associated with A20 diffuse large B-cell lymphoma (DLBCL) cells seeding of different tissue microenvironments. Furthermore, we characterized features of lymphoma-associated stromatogenesis in human DLBCL samples using Digital Spatial Profiling, and established their relationship with prognostically relevant variables, such as MYC. Findings: We found that the tissue microenvironment casts a relevant influence over A20 transcriptional landscape also impacting on Myc and DNA damage response programs. Extending the investigation to mice deficient for the matricellular protein SPARC, a stromal prognostic factor in human DLBCL, we demonstrated a different immune imprint on A20 cells according to stromal Sparc proficiency. Through Digital Spatial Profiling of 87 immune and stromal genes on human nodal DLBCL regions characterized by different mesenchymal composition, we demonstrate intra-lesional heterogeneity arising from diversified mesenchymal contextures and impacting on the stromal and immune milieu. Interpretation: Our study provides experimental evidence that stromal microenvironment generates topological determinants of intra-tumour heterogeneity in DLBCL involving key transcriptional pathways such as Myc expression, damage response programs and immune checkpoints. Funding: This study has been supported by the Italian Foundation for Cancer Research (AIRC) (grants 15999 and 22145 to C. Tripodo) and by the University of Palermo.
