Type: Oral
Session: 605. Molecular Pharmacology and Drug Resistance: Lymphoid Neoplasms: Mechanisms of Therapy Resistance in Lymphoma
Hematology Disease Topics & Pathways:
Combination therapy, Apoptosis, Lymphomas, Non-Hodgkin lymphoma, B Cell lymphoma, Diseases, Treatment Considerations, Immunology, Lymphoid Malignancies, Biological Processes
Methods: We developed tet-on expression systems of BCL10 gain-of-function mutants (R58Q and S136X). We examined gene-expression profiling (GEP), cytokine arrays, and transcription factor enrichment by ENCODE ChIP-X Enrichment Analysis Version 3 (ChEA3). We employed drug screening to interrogate impacts on a variety of therapeutic classes using epigenetic Library (TargetMol). We examined activities of a drugs alone and in combination in vitro by measuring cell viability and in xenograft models (NOD scid gamma mice) by measuring tumor volume post treatments. We engineered the activating BCL10-S136X truncation as a cre-inducible conditional allele at the murine Rosa26 locus (called RSB mice) and crossed it later with the IµBCL6 transgenic background to generate an accurate immunocompetent BN2/C1 mouse model.
Results: GEP revealed reprogramming converging on NF-kB activation reinforced by cytokines (CCL17, CCL4, IL32, CXCL10, TNFa, TNFb). Cytokines upregulated by the mutants were also significantly upregulated in BCL10-mutant patient tumors including CCL22 (p=0.0011) and IL7 (p=0.0013). Further, ELISA confirmed the activation of TNFb (p<0.01) and TNFa( p<0.01). Drug screening revealed numerous additional classes besides BTKis whose activities are thwarted by BCL10-mutant activities. These include the BCL2 inhibitor venetoclax, an FDA-approved drug with remarkable activities in other B-cell non-Hodgkin lymphomas but largely inactive in DLBCL. We also found in BCL10-mutant samples significantly higher expression of BCL2L1 (S136X p=0.0001, R58Q P=0.0002), BCL2A1(S136X p=0.0011, R58Q p=0.0005), and BCL2 itself (S136X p=0.0001, R58Q p=0.0074). We find the BCL10 mutant-driven cytokine-reinforced positive feedback loop of lymphomagenesis activates multiple additional pathways converging on diffuse activation of oncogenic transcription factors. BCL10 truncation mediated up-regulation of anti-apoptotic genes promotes increased mitochondrial membrane potential, a key factor underlying multidrug, including venetoclax, resistance, but further investigation revealed this is overcome by the addition of the potent non-covalent BTK inhibitor pirtobrutinib. 24 hours of pirtobrutinib treatment drastically reduced the transcript levels of BCL2 family genes in BCL10 mutants comparing to vector; BCL2 (vector: p=0.0325, R58Q: p=0.005, S136X: p=0.0006), BCL2L1 (vector: p=ns, R58Q: p=0.0484, S136X: p=0.0007), BCL2A1 (vector: p=0.0087, R58Q: p=0.0121, S136X: p=0.0005). Venetoclax+pirtobrutinib synergized and potently killed BCL10-mutant tumors in vitro (Bliss synergy score_ RIVA; vector =7.96, R58Q=9.03, S136X=13.82) and in vivo. RSB x MB-1/cre crosses did not impact survival of double-het offspring but promoted an activation block thwarting germinal center responses to antigen. Crossing these mice into the IµBCL6 transgenic background that replicates a defining chromosomal translocation of BN2/C1 cases resulted in tumors resembling DLBCL or marginal zone lymphoma with 100% penetrance and compromised overall survival (p<0.0001, Mantel-Cox test).
Conclusion: The combination of venetoclax and pirtobrutinib synergistically overcomes BCL10-driven resistance and effectively kills BCL10-mutant DLBCL tumors in vitro and in vivo, suggesting BTK retains key roles protecting DLBCL tumors from apoptosis even when dispensable for CBM activation. Genetically accurate mutant-BCL10 mice revealed a novel germinal-center activation deficit and facilitated generation of valuable preclinical models to further interrogate biology of the BN2/C1 molecular subgroup through activation of the BCL6 oncogene.
Disclosures: Landgren: Amgen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees; Merck: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees; Theradex: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees.; Takeda: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees; Janssen: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees; Celgene: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees; Adaptive: Consultancy, Honoraria, Membership on an entity's Board of Directors or advisory committees, Other: Membership on independent data monitoring committees. Maura: Sanofi: Consultancy, Honoraria; Medidata: Consultancy, Honoraria.
See more of: Oral and Poster Abstracts