Mantle cell lymphoma (MCL) is a hematologic neoplasm characterised by the t(11;14)(q13;q32) translocation leading to aberrant cyclin D1 expression. The cell functions of cyclin D1 depend on its partners and/or subcellular distribution, resulting in different oncogenic properties. We observed the accumulation of cyclin D1 in the cytoplasm of a subset of MCL cell lines and primary cells. In primary cells, this cytoplasmic distribution was correlated with a more frequent blastoid phenotype. We performed immunoprecipitation assays and mass spectrometry on enriched cytosolic fractions from two cell lines. The cyclin D1 interactome was found to include several factors involved in adhesion, migration and invasion. We found that the accumulation of cyclin D1 in the cytoplasm was associated with higher levels of migration and invasiveness. We also showed that MCL cells with high cytoplasmic levels of cyclin D1 engrafted more rapidly into the bone marrow, spleen, and brain in immunodeficient mice. Both migration and invasion processes, both in vivo and in vitro, were counteracted by the exportin 1 inhibitor KPT-330, which retains cyclin D1 in the nucleus. Our data reveal a role of cytoplasmic cyclin D1 in the control of MCL cell migration and invasion, and as a true operator of MCL pathogenesis.

Constitutive activation of the chemokine receptor CXCR4 has been associated with tumor progression, invasion, and chemotherapy resistance in different cancer subtypes. Although the CXCR4 pathway has recently been suggested as an adverse prognostic marker in diffuse large B-cell lymphoma, its biological relevance in this disease remains underexplored. In a homogeneous cohort of 52 patient biopsies, an antibody-based cytokine array showed that tissue levels of CXCL12 correlated with high microvessel density and bone marrow involvement at diagnosis, supporting a role for CXCL12-CXCR4 axis in disease progression. We then identified the tetra-amine IQS-01.01RS as a potent inverse agonist of the receptor, preventing CXCL12-mediated chemotaxis and triggering apoptosis in a panel of 18 cell lines and primary cultures, with superior mobilizing properties in vivo than standard agent. IQS-01.01RS activity was associated with downregulation of p-AKT, p-ERK1/2 and destabilization of MYC, allowing to a synergistic interaction with the bromodomain and extra-terminal domain inhibitor, CPI203. In a xenotransplant model of diffuse large B-cell lymphoma, IQS-01.01RS/CPI203 combination decreased tumor burden through MYC and p-AKT downregulation, and enhanced apoptosis induction. Thus, our results point out an emerging role of CXCL12-CXCR4 in the pathogenesis of diffuse large B-cell lymphoma and support the simultaneous targeting of CXCR4 and bromodomain proteins as a promising, rationale-based strategy for this disease.

Staphylococcus aureus is a commensal bacterium that causes severe infections in soft tissue and the bloodstream. During infection, S. aureus manipulates host cell response to facilitate its own replication and dissemination. Here, we show that S. aureus significantly decreases the level of SUMOylation, an essential post-translational modification, in infected macrophages 24 h post-phagocytosis. The reduced level of SUMOylation correlates with a decrease in the SUMO-conjugating enzyme Ubc9. The over-expression of SUMO proteins in macrophages impaired bacterial intracellular proliferation and the inhibition of SUMOylation with ML-792 increased it. Together, these findings demonstrated for the first time the role of host SUMOylation response toward S. aureus infection.

A significant proportion of diffuse large B cell lymphoma (DLBCL) and follicular lymphoma (FL) patients harbor a gain-of-function, heterozygous somatic mutations of the methyltransferase gene EZH2. Despite acceptable safety profile and early signs of activity in clinical trials, single agent treatment with EZH2 inhibitors is unlikely to be curative in aggressive lymphomas. In an effort to established novel rational combinations, we have evaluated the activity and mechanism of action of the EZH2 small molecule inhibitor CPI169 as single agent and in combination with BET bromodomain inhibition, using preclinical models of DLBCL and FL with distinct EZH2 mutational status. CPI169 anti-tumor activity and specificity was assessed in vitro in 10 DLBCL and FL cell lines, including cells expressing basal or ectopic EZH2mut. Molecular bases of its activity were determined by gene expression profiling (GEP), qPCR and western blot, followed by automated exploratory data analysis. Biomarkers validation was made in vitro and in vivo, using a mouse xenotransplant model of EZH2mut DLBCL, considering both exposure to CPI169 single agent and/or combination treatment with a BRD4 inhibitor, CPI203. CPI169 induced dose-dependent proliferation blockade in EZH2mut, but not EZH2wt DLBCL and FL cell lines, independently of EZH2 expression level or basal methyltransferase activity. Loss of H3K27me3 mark upon CPI169 treatment was associated with upregulation of gene sets related to G1 cell cycle blockade, mTOR and P53 pathways, and MYC signaling. Accordingly, combination with the MYC-interfering drug, BET inhibitor CPI203, achieved a synergistic anti-proliferating activity in EZH2 mutated cases and in mice bearing EZH2mut DLBCL tumors. Activity of EZHi/BRD4i combo was characterized by lower mitotic index, increased loss of H3K27me3 mark, in association with MYC downregulation. GEP analysis, followed by automated exploratory data analysis and validation by a siRNA screening, further identified the PI3K/AKT-regulated gene and mitosis regulator, YPEL2, and the regulator of innate-like B lymphocyte maturation, KLHL14, as crucial factors involved in the efficacy of MYC/EZH2 dual targeting. In conclusion, CPI169 shows significant activity and safety as single agent in EZH2 mutated-DLBCL and FL cases and displays synergistic interaction in vitro and in vivo with BRD4 inhibition, mediated by the modulation of a limited set of EZH2-regulated genes. Citation Format: Aranzazu Chamorro-Jorganes, Marcelo L. Ribeiro, Nuria Profitos-Peleja, Diana Reyes-Garau, Clara Recasens-Zorzo, Juan G. Valero, Marc Armengol, Patricia Perez-Galan, Ray Butler, Antonio Postigo, Francesc Bosch, Gael Roue. Safety and efficacy of EZH2 and BRD4 dual targeting in EZH2Y641mut germinal centre-derived lymphoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2925.