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453 B Cell Receptor Silencing Reveals Origin and Impacts Treatment of High-Grade B Cell Lymphomas with MYC and BCL2 Rearrangements

Program: Oral and Poster Abstracts
Type: Oral
Session: 621. Lymphomas: Translational – Molecular and Genetic: Single-cell and Spatial Analyses in Aggressive and T Cell Lymphomas
Hematology Disease Topics & Pathways:
Research, Translational Research
Sunday, December 8, 2024: 10:00 AM

Stefano Casola, MD, PhD1,2, Paola Sindaco, MD3, Silvia Lonardi4*, Gabriele Varano, PhD5, Ilaria Pietrini6*, Gaia Morello7*, Piera Balzarini8*, Hadas Neuman9*, Filippo Vit, MSc10*, Giorgio Bertolazzi11*, Hiroshi Arima, MD, PhD6,12*, Marco Chiarini, PhD13*, Luisa Lorenzi8*, Vilma Pellegrini14*, Sabrina Giampaolo6*, Daniel Garzon6*, Cecilia Ranise6*, Mattia Bugatti15*, Chiara Pagani, MD16*, Rosa Daffini, MD17*, Federica Mainoldi6*, Viveka Selvarasa6*, Anojan Sivacegaram6*, Henry Yang18*, Li Ying18*, Valeria Cancila7*, Raoul Bonnal19*, Euplio Visco20*, Cristina Lopez Gonzalez21,22*, Pasquale Capaccio23*, Andrés José María Ferreri, MD24, Alessandra Tucci, MD25, Antonello Cabras26*, Giancarlo Pruneri, MD27*, Arianna Di Napoli, MD28*, Reiner Siebert, PhD29*, Riccardo Bomben, PhD30*, Brunangelo Falini31, Marco Pizzi32*, Joo Y. Song, MD33, Wing Chung Chan, MD34, Maurilio Ponzoni, MD35, Ramit Mehr36*, Claudio Tripodo11* and Fabio Facchetti37*

1IFOM-ETS, Milan, Italy
2Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan, Italy
3Inselspital, Universitätsspital Bern, Department of Hematology and Central Hematology Laboratory., Bern, Switzerland
4Spedali Civili Brescia, Volta Mantovana, MN, ITA
5IFOM-ETS, Milano, Italy
6IFOM ETS-The AIRC Institute of Molecular Oncology, Milano, Italy
7University of Palermo, Palermo, ITA
8Spedali Civili Di Brescia, Brescia, ITA
9Bar Ilan University, The Mina and Everard Goodman Faculty of Life Sciences, Ramat Gan, Israel
10Clinical and Experimental Onco-Haematology Unit, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, Aviano, Italy, Italy
11University of Palermo, Palermo, Italy
12Kyoto University, Kyoto, JPN
13Clinical Chemistry Laboratory, Diagnostic Department, ASST Spedali Civili Di Brescia, Brescia, Italy
14Pathology unit; University of Brescia, Spedali Civili, Department of Molecular and Translational Medicine, Brescia, Italy, Brescia, Italy
15BRESCIA UNIVERSITY, BRESCIA, ITA
16ASST Spedali Civili di Brescia, Brescia, Italy
17Hematology, ASST Spedali Civili di Brescia, Brescia, Italy
18Cancer Science Institute of Singapore, National University of Singapore, Singapore, Singapore
19The AIRC Institute of Molecular Oncology (IFOM), Milan, Italy
20Pathology unit, IRCCS San Raffaele Scientific Institute, Vita-Salute San Raffaele University, Milano, Italy
21University Medical Center JGU Mainz, Mainz, DEU
22Hematopathology Unit, Hospital Clínic Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS),, Barcelona, Spain
23University of Milan, Milano, Italy
24Unit of Lymphoid Malignancies, Department of Onco-Haematology, IRCCS San Raffaele Scientific Institute, Milano, Italy
25UOC Ematologia, Brescia, Italy
26Istituto Nazionale Tumori, Milan, Mi, ITA
27Istituto Nazionale dei Tumori Foundation, Milan, Italy
28Sapienza University of Rome-sant'Andrea Hospital, Rome, ITA
29Institute of Human Genetics, Ulm University and Ulm University Medical Center, Ulm, Ulm, DEU
30Clinical and Experimental Onco-Hematology Unit, Centro di Riferimento Oncologico di Aviano (CRO) IRCCS, Aviano, Italy
31Institute of Hematology and Center for Hemato-Oncology Research, University and Hospital of Perugia, Perugia, Italy
32University of Padua, Padova, ITA
33Department of Hematology and Hematopoietic Cell Transplantation, City of Hope National Medical Center, Duarte, CA
34Department of Pathology, City of Hope National Medical Center, Duarte, CA
35Pathology Unit, IRCCS San Raffaele Scientific Institute and Università Vita-Salute San Raffaele, Milan, Italy
36Bar-Ilan University, Ramat-Gan, ISR
37University of Brescia, Brescia, ITA

High-grade B cell lymphomas with MYC and BCL2 rearrangements, also referred as MYC/BCL2 double-hit lymphomas (HGBCL-DH-BCL2), represent highly aggressive B cell malignancies with dismal prognosis and limited therapeutic options. Beside the suggested origin from germinal center (GC) B cells, pathogenesis of HGBCL-DH-BCL2 remains largely unknown.

The B cell receptor (BCR) represents a major signaling hub and preferred therapeutic target for several mature B cell lymphoma/leukemias. We investigated expression of BCR complex components, including immunoglobulin (IG) heavy (H) and light (L) chains, and CD79A and CD79B signaling subunits in 93 HGBCL-DH-BCL2 cases. In 66% of the tumors, undetectable IGM/D/G/A IGH (IGHUND) protein assessed by immunohistochemistry, flow cytometry and proximity ligation assays was linked to CD79B downregulation and intracellular retention. Remaining 34% predominantly expressed surface (s) IGM/CD79B/CD79A+ BCR complexes. Bulk and spatial transcriptomics identify IGHUND and IGH+ HGBCL-DH-BCL2 lymphomas as distinct biological entities, with the former resembling GC DZ B cells limiting T cell recall. Instead, IG+ HGBCL-DH-BCL2 established an immune/stromal cell-rich microenvironment permetated by immunosuppressive cell types and soluble signals, with transcriptional similarities to GC light zone-resident B cells. IGHUND HGBCL-DH-BCL2 selected significantly higher c-MYC expression than their IGH+ counterparts and boosted mitochondrial respiration. IGHUND and IGH+ tumors shared non-synonymous single-nucleotide variants in several genes recurrently mutated in classical Follicular Lymphoma (FL), while selecting subset-specific gene mutations indicative of distinct evolutionary trajectories. IGHUND HGBCL-DH-BCL2 preserved potentially productive, hypermutated IGH variable (V) gene rearrangements, counterselecting structure-damaging aminoacid replacements, while bypassing antigen-driven selection. IGHUND HGBCL-DH-BCL2 originated from IGG/IGA class switched GC B cells, while IGH-expressing tumors were selected for continuous IGM expression. IGH analyses in 13 metachronous tumor pairs reveal that IGH shutdown and class choice are pre-determined in the FL preceding HGBCL-DH-BCL2. Mutational analyses on metachronous cases indicate that HGBCL-DH-BCL2 can either evolve from FL, or arise from a FL/HGBCL-DH-BCL2 common precursor cell (CPC) through branched evolution. IGHUND HGBCL-DH-BCL2 showed recurrent (43%) bi-allelic disruption of the IGH locus, which only occasionally intercepted the MYC locus. Sequencing of the t(8;22) translocation breakpoint in an IGHUND HGBCL-DH-BCL2 case, revealed juxtaposition of MYC to the IG lambda (L) J2 gene segment, replacing recombination signal sequences, consistent with illegitimate RAG1/2-dependent VJ recombination. In support of this, lymphoma cells showed widespread RAG1/2 expression, with similar results confirmed in 26 % of additional HGBCL-DH-BCL2 cases, and in corresponding t(8;22)+ cell lines, which commonly lacked sIG expression. We show that in HGBCL-DH-BCL2, RAG1/2 (re-)expression is facilitated by BCR downregulation, recalling mechanisms of BCR revision/editing promoting secondary IG light chain V gene rearrangements. In line with this, IGHUND HGBCL-DH-BCL2 showed a striking bias for IG lambda light chain usage, at the expense of IG kappa chains, which were predominant among the IGH+ counterparts. Importantly, patient-derived IGHUND HGBCL-DH-BCL2 lymphoma cell lines showed in vitro resistance to the sCD79B-targeting antibody-drug conjugate Polatuzumab-Vedotin (PV). In conclusion, we report, for the first time, the predominant shutdown of sBCR expression in HGBCL-DH-BCL2. BCR silencing, starting in the t(14;18)+ FL/CPC cell triggers RAG1/2-dependent BCR editing, which favors t(8;22), IGL::MYC rearrangements, and, possibly non IG-MYC translocations, ultimately driving transformation into IGHUND HGBCL-DH-BCL2. Our results reconcile with the reduced efficacy of PV in the treatment of HGBCL-DH-BCL2, urging for routine surface CD79B assessment to guide treatment indication.

Disclosures: Arima: Verastem, Inc: Other. Tucci: Kiowa Kyrin: Membership on an entity's Board of Directors or advisory committees; Lilly: Membership on an entity's Board of Directors or advisory committees; Incyte: Membership on an entity's Board of Directors or advisory committees; Regeneron: Membership on an entity's Board of Directors or advisory committees; Beigene: Membership on an entity's Board of Directors or advisory committees; Takeda: Membership on an entity's Board of Directors or advisory committees; Abbvie: Membership on an entity's Board of Directors or advisory committees; Sanofi: Membership on an entity's Board of Directors or advisory committees; Gentili: Membership on an entity's Board of Directors or advisory committees.

*signifies non-member of ASH