Aggressive B-cell lymphomas are clinically and pathologically diverse and reflect multiple pathways of transformation involving, among others, oncogenic signaling, lymphoma-stroma crosstalk and intrinsic protein homeostasis. Alterations in these highly regulated processes play a key role in the progression of the malignant clone and correlate with a high failure rate in treatment protocols. In the last decade, new therapies applied to the treatment of B-cell lymphoma have significantly improved the overall survival of these patients, but at present no drug can cure these diseases. At the origin of the gap between promising preclinical results and failure in clinical phase II/III, conventional preclinical models lack predictive value in the main trials carried out in these cancer subtypes. For the design of new therapies more selective and more suited to the biology of these lymphomas, our team focuses on the characterization of new drug targets and on the bench-to-bedside translation of innovative therapeutic strategies, using innovative experimental models of diffuse large B cell lymphoma (DLBCL), mantle cell lymphoma (MCL) and transformed follicular lymphoma (tFL).
Development of a patient-derived xenografts platform for the evaluation of new targeted therapies in aggressive B-cell lymphomas
Much of the new therapies applied to the treatment of aggressive B-cell lymphomas have not improved the overall survival of patients and there is no drug or drug combinations to cure these diseases. To counteract the resistance and toxicity observed in the clinic, novel and personalized approaches, new preclinical models that allow predicting more effectively the specificity and safety of each treatment, are urgently needed. Current therapeutic targets of highest interest in these entities include distinct signaling pathways that are activated constitutively by over-expression of MYC, BCL6, or CRBN, activation of phosphatidylinositol 3-kinase (PI3K) and B-cell receptor (BCR)-related kinases, or by the deregulation of apoptotic program. Currently, there are several therapeutic agents directed specifically towards these signaling axes, although their efficacy, safety and mode of action are still to be determined in some subtypes of B-cell lymphoma. To warrant the efficacy, safety and translationnality of these studies, we propose to associate standard in vitro assays with innovative in vivo models (PDX) that will allow us 1) to work directly with primary tumor cells, validating the most effective therapies and their biological effects with greater relevance, and taking into account the role of the tumor microenvironment and 2) to guarantee the possible translation of these therapies into clinical trials with the molecular and genetic determination of the factors that condition the response to the best therapies tested in each patient. The collaboration with pharmaceutical companies will allow the transfer of the results obtained to Hematology Departments for the design of new phase I/II clinical trials.
Modulation of lymphoid microenvironment by intrinsic protein homeostasis in aggressive B-cell lymphoma
The lack of efficacy of standard and experimental therapies in the clinical is likely due to the uncontrolled activity of some components of the tumor microenvironment (TME), including tumor-associated macrophages, mesenchymal cells, dendritic cells, NK cells or regulatory T cells. Although major advances have been done in the last decade about the role of accompanying immune effectors in the control of B-NHL tumor growth and resistance to standard and experimental therapies, how MCL and DLBCL malignant B cells modulate their environment to better adapt to adverse conditions, is poorly understood. Accumulating evidences in solid cancers suggest that increased expression and/or activity of the ubiquitin-proteasome system, including Ub and Ub-like protein modifiers, may modulate some TME components indirectly, thus promoting an immunosuppressive, protumoral environment. However, little is known about the relevance of the intrisinc tumoral Ub interactome (ubiquitome) in the bidirectional tumor-stroma crosstalk in MCL and DLBCL. From a therapeutic point of view, this field of research may be of special interest, as a number of new agents directed against proteasome, ubiquitin or ubiquitin-like ligases or deubiquitinase enzymes, are showing promising preclinical and clinical activity in MCL and DLBCL patients. The discovery of new Ub-regulated factors favoring or counteracting the recruitment and/or activation of immune effector cells, may pave the way for the comprehension of tumor-stroma interplay in MCL and DLBCL and for the design of new, rationally-based therapeutic approaches combining post-translational modifying drugs and immunotherapeutic agents.
The main objective of this project is the identification of new mechanisms related to intrinsic protein homeostasis that may regulate the complex interplay between MCL and DLBCL cells and their specific TME, using experimental models with the capacity to preserve the spatial architecture of the original tumor. The specific objectives are 1) to generate a collection of in vitro (3D organoid) and in vivo (PDX) models from MCL/DLBCL fresh samples able to maintain the spatial organization and the cell-cell interaction established in the original tumor. These experimental models will be fully characterized immunophenotypically for their content in immune effector cells; 2) to characterize the ubiquitome of malignant B cells by proteomic profiling and to correlate these intracellular complexes with the immunological pattern of each tumor model; 3) to validate the impact of tumor protein homeostasis on the development of B-cell lymphoma and on the intratumoral infiltration of immune cell, by automatic data analysis and CRISPR-Cas9-mediated gene silencing in DLBCL and MCL 3D organoid models.
- Manuel S. Rodriguez, ITAV CNRS USR3505, Toulouse, France
- Rosa Barrio, Ubiquitin-likes And Development Lab, CICBioGUNE, Derio, Spain
- Laurent Poulain, Inserm U1086 « ANTICIPE », Université de Caen Normandie, Caen, France
- Ray G. Butler, Butler Scientifics, Barcelona, Spain
Show all publications
Pharmacological modulation of CXCR4 cooperates with BET bromodomain inhibition in diffuse large B-cell lymphoma.Haematologica Abr 2019, 104 (4) 778-788. Epub 28 Jun 2018
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 set of 52 biopsies from patients, 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 the 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 those of the standard agent. IQS-01.01RS activity was associated with downregulation of p-AKT, p-ERK1/2 and destabilization of MYC, allowing a synergistic interaction with the bromodomain and extra-terminal domain inhibitor, CPI203. In a xenotransplant model of diffuse large B-cell lymphoma, the combination of IQS-01.01RS and CPI203 decreased tumor burden through MYC and p-AKT downregulation, and enhanced the induction of apoptosis. 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 the treatment of this disease.Més informació
The BET bromodomain inhibitor CPI203 overcomes resistance to ABT-199 (venetoclax) by downregulation of BFL-1/A1 in in vitro and in vivo models of MYC+/BCL2+ double hit lymphoma.Oncogene Abr 2018, 37 (14) 1830-1844. Epub 22 Gen 2018
High-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements, mostly known as double-hit lymphoma (DHL), is a rare entity characterized by morphologic and molecular features between Burkitt lymphoma and the clinically manageable diffuse large B-cell lymphoma (DLBCL). DHL patients usually undergo a rapidly progressing clinical course associated with resistance to standard chemo-immunotherapy. As a consequence, the prognosis of this entity is particularly poor with a median overall survival inferior to 1 year. ABT-199 (venetoclax) is a potent and selective small-molecule antagonist of BCL-2 recently approved for the treatment of a specific subtype of lymphoid neoplasm. In this study, we demonstrate that single-agent ABT-199 efficiently displaces BAX from BCL-2 complexes but fails to maintain a significant antitumor activity over time in most MYC+/BCL2+DHL cell lines and primary cultures, as well as in a xenograft mouse model of the disease. We further identify the accumulation of the BCL2-like protein BFL-1 to be a major mechanism involved in acquired resistance to ABT-199. Noteworthy, this phenomenon can be counteracted by the BET bromodomain inhibitor CPI203, since gene expression profiling identifies BCL2A1, the BFL-1 coding gene, as one of the top apoptosis-related gene modulated by this compound. Upon CPI203 treatment, simultaneous downregulation of MYC and BFL-1 further overcomes resistance to ABT-199 both in vitro and in vivo, engaging synergistic caspase-mediated apoptosis in DHL cultures and tumor xenografts. Together, these findings highlight the relevance of BFL-1 in DH lymphoma-associated drug resistance and support the combined use of a BCL-2 antagonist and a BET inhibitor as a promising therapeutic strategy for patients with aggressive DHL.Més informació
Activity of the novel BCR kinase inhibitor IQS019 in preclinical models of B-cell non-Hodgkin lymphoma.J Hematol Oncol 31 Mar 2017, 10 (1) 80. Epub 31 Mar 2017
Pharmacological inhibition of B cell receptor (BCR) signaling has recently emerged as an effective approach in a wide range of B lymphoid neoplasms. However, despite promising clinical activity of the first Bruton's kinase (Btk) and spleen tyrosine kinase (Syk) inhibitors, a small fraction of patients tend to develop progressive disease after initial response to these agents.Més informació
Synergistic antitumor activity of lenalidomide with the BET bromodomain inhibitor CPI203 in bortezomib-resistant mantle cell lymphoma.Leukemia Oct 2014, 28 (10) 2049-59. Epub 18 Mar 2014
Bortezomib therapy has shown promising clinical activity in mantle cell lymphoma (MCL), but the development of resistance to proteasome inhibition may limit its efficacy. To unravel the factors involved in the acquisition of bortezomib resistance in vivo, immunodeficient mice were engrafted with a set of MCL cell lines with different levels of sensitivity to the drug, followed by gene expression profiling of the tumors and functional validation of the identified gene signatures. We observed an increased tumorigenicity of bortezomib-resistant MCL cells in vivo, which was associated with plasmacytic differentiation features, like interferon regulatory factor 4 (IRF4) and Blimp-1 upregulation. Lenalidomide was particularly active in this subgroup of tumors, targeting IRF4 expression and plasmacytic differentiation program, thus overcoming bortezomib resistance. Moreover, repression of the IRF4 target gene MYC in bortezomib-resistant cells by gene knockdown or treatment with CPI203, a BET (bromodomain and extra terminal) bromodomain inhibitor, synergistically induced cell death when combined with lenalidomide. In mice, addition of CPI203 to lenalidomide therapy further decreased tumor burden, involving simultaneous MYC and IRF4 downregulation and apoptosis induction. Together, these results suggest that exacerbated IRF4/MYC signaling is associated to bortezomib resistance in MCL in vivo and warrant clinical evaluation of lenalidomide plus BET inhibitor combination in MCL cases refractory to proteasome inhibition.Més informació
The Hsp90 inhibitor IPI-504 overcomes bortezomib resistance in mantle cell lymphoma in vitro and in vivo by down-regulation of the prosurvival ER chaperone BiP/Grp78.Blood 27 Gen 2011, 117 (4) 1270-9. Epub 24 Nov 2010
Despite the promising introduction of the proteasome inhibitor bortezomib in the treatment of mantle cell lymphoma (MCL), not all patients respond, and resistance often appears after initial treatment. By analyzing a set of 18 MCL samples, including cell lines with constitutive or induced resistance to bortezomib, we found a high correlation between loss of sensitivity to the proteasome inhibitor and up-regulation of the prosurvival chaperone BiP/Grp78. BiP/Grp78 stabilization was ensured at a posttranscriptional level by an increase in the chaperoning activity of heat shock protein of 90 kDa (Hsp90). In bortezomib-resistant cells, both BiP/Grp78 knockdown and cell pretreatment with the Hsp90 inhibitor of the ansamycin class, IPI-504, led to synergistic induction of apoptotic cell death when combined with bortezomib. Cell exposure to the IPI-504-bortezomib combination provoked the dissociation of Hsp90/BiP complexes, leading to BiP/Grp78 depletion, inhibition of unfolded protein response, and promotion of NOXA-mediated mitochondrial depolarization. The IPI-504-bortezomib combination also prevented BiP/Grp78 accumulation, thereby promoting apoptosis and inhibiting the growth of bortezomib-resistant tumors in a mouse model of MCL xenotransplantation. These results suggest that targeting unfolded protein response activation by the inhibition of Hsp90 may be an attractive model for the design of a new bortezomib-based combination therapy for MCL.Més informació
Development of a patient-derived xenografts platform for the evaluation
|Data de finalització:||31/12/2021|
Evaluation of the ”U2” drug combo in association with anti-PD-L1 antibody, BET inhibitor TG-1601, BTK inhibitor TG-1701, in preclinical models of B-cell non-Hodgkin lymphoma (B-NHL)
|Data de finalització:||30/06/2020|
Evaluation of a new class of cereblon modulators in preclinical models of diffuse large B cell lymphoma
|Data de finalització:||31/12/2019|
Modulation of lymphoid microenvironment by intrinsic protein homeostasis in aggressive B-cell lymphoma
|Data de finalització:||31/12/2021|