In vivo Model of Follicular Lymphoma Resistant to Rituximab

Purpose: Follicular lymphoma (FL) is the most common subtype of indolent lymphomas. Rituximab is widely used alone or in combination therapy for the treatment of FL. Despite its well-established clinical efficacy, a subpopulation of patients does not respond to rituximab and most patients will relapse after therapy. The mechanisms of action and resistance to rituximab are not fully understood. Experimental Design: To study these mechanisms we developed an in vivo model of FL resistant to rituximab. This model was developed using the human RL line, isolated from a patient with FL, grown as xenotransplants in severe combined immunodeficient mice, exposed weekly to rituximab in vivo, followed by serial reimplantation and reexposure to rituximab, until a resistant phenotype was obtained. Results: RL-derived tumors unexposed to rituximab were grown as controls and compared with the resistant tumors. Although the expression of CD46 and CD55 antigens were not differently expressed in the resistant cells, the complement inhibitor CD59 was overexpressed in a subpopulation and CD20 was found to be expressed at a lower level in a minority of cells. Bcl-XL and YY1 were also found more highly expressed in rituximab-resistant cells. Conclusion: This model provides insight on potential in vivo resistance mechanisms to rituximab and could help contribute to the development of novel therapies in rituximab-refractory diseases.

[1]  J. Wilde,et al.  Rituxan (anti‐CD20 antibody)‐induced translocation of CD20 into lipid rafts is crucial for calcium influx and apoptosis , 2005, Clinical and experimental immunology.

[2]  C. Dumontet,et al.  A20/TNFAIP3, a new estrogen-regulated gene that confers tamoxifen resistance in breast cancer cells , 2007, Oncogene.

[3]  N. Van Rooijen,et al.  The role of complement in the therapeutic activity of rituximab in a murine B lymphoma model homing in lymph nodes. , 2006, Haematologica.

[4]  J. Hainsworth,et al.  Rituximab as first-line and maintenance therapy for patients with indolent non-hodgkin's lymphoma. , 2002, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[5]  N. Schmitz,et al.  Frontline therapy with rituximab added to the combination of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) significantly improves the outcome for patients with advanced-stage follicular lymphoma compared with therapy with CHOP alone: results of a prospective randomized study of t , 2005, Blood.

[6]  Valeria Grieco,et al.  Complement Activation Determines the Therapeutic Activity of Rituximab In Vivo 1 , 2003, The Journal of Immunology.

[7]  R. Gressin,et al.  In vitro mechanisms of action of rituximab on primary non-Hodgkin lymphomas. , 2003, Blood.

[8]  R. Levy,et al.  Rituximab chimeric anti-CD20 monoclonal antibody therapy for relapsed indolent lymphoma: half of patients respond to a four-dose treatment program. , 1998, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[9]  D. Longo,et al.  Phorbol ester-induced, cell-cycle-specific, growth inhibition of human B-lymphoma cell lines. , 1990, Journal of the National Cancer Institute.

[10]  A. Ho,et al.  Level of CD 20-expression and Efficacy of Rituximab Treatment in Patients with Resistant or Relapsing B-cell Prolymphocytic Leukemia and B-cell Chronic Lymphocytic Leukemia , 2002, Leukemia & lymphoma.

[11]  A. López-Guillermo,et al.  Complement-mediated cell death induced by rituximab in B-cell lymphoproliferative disorders is mediated in vitro by a caspase-independent mechanism involving the generation of reactive oxygen species. , 2001, Blood.

[12]  B. Coiffier.  Rituximab therapy in malignant lymphoma , 2007, Oncogene.

[13]  B. Bonavida,et al.  ‘Rituximab-induced inhibition of antiapoptotic cell survival pathways: implications in chemo/immunoresistance, rituximab unresponsiveness, prognostic and novel therapeutic interventions’ , 2007, Oncogene.

[14]  Mitchell R. Smith.  Rituximab (monoclonal anti-CD20 antibody): mechanisms of action and resistance , 2003, Oncogene.

[15]  S. de Vos,et al.  Rituximab (anti-CD20) selectively modifies Bcl-xL and apoptosis protease activating factor-1 (Apaf-1) expression and sensitizes human non-Hodgkin's lymphoma B cell lines to paclitaxel-induced apoptosis. , 2003, Molecular cancer therapeutics.

[16]  M. Czuczman,et al.  Acquirement of Rituximab Resistance in Lymphoma Cell Lines Is Associated with Both Global CD20 Gene and Protein Down-Regulation Regulated at the Pretranscriptional and Posttranscriptional Levels , 2008, Clinical Cancer Research.

[17]  B. Bonavida,et al.  Inhibition of interleukin 10 by rituximab results in down-regulation of bcl-2 and sensitization of B-cell non-Hodgkin's lymphoma to apoptosis. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[18]  G. Cheng,et al.  Rituximab (chimeric anti-CD20 monoclonal antibody) inhibits the constitutive nuclear factor-{kappa}B signaling pathway in non-Hodgkin's lymphoma B-cell lines: role in sensitization to chemotherapeutic drug-induced apoptosis. , 2005, Cancer research.

[19]  M. Czuczman,et al.  Acquired Resistance to Rituximab Is Associated with Chemotherapy Resistance Resulting from Decreased Bax and Bak Expression , 2008, Clinical Cancer Research.

[20]  G. Schieven,et al.  Association of tyrosine and serine kinases with the B cell surface antigen CD20. Induction via CD20 of tyrosine phosphorylation and activation of phospholipase C-gamma 1 and PLC phospholipase C-gamma 2. , 1993, Journal of immunology.

[21]  L. Presta,et al.  Inhibitory Fc receptors modulate in vivo cytoxicity against tumor targets , 2000, Nature Medicine.

[22]  J. Byrd,et al.  Rituximab in B-cell chronic lymphocytic leukemia. , 2003, Seminars in oncology.

[23]  G. Salles,et al.  Rituximab (anti-CD20 monoclonal antibody) as single first-line therapy for patients with follicular lymphoma with a low tumor burden: clinical and molecular evaluation. , 2001, Blood.

[24]  B. Dörken,et al.  Anti-CD20- and B-cell receptor-mediated apoptosis: evidence for shared intracellular signaling pathways. , 2000, Cancer research.

[25]  J. Byrd,et al.  The mechanism of tumor cell clearance by rituximab in vivo in patients with B-cell chronic lymphocytic leukemia: evidence of caspase activation and apoptosis induction. , 2002, Blood.

[26]  J. Hainsworth,et al.  Maximizing therapeutic benefit of rituximab: maintenance therapy versus re-treatment at progression in patients with indolent non-Hodgkin's lymphoma--a randomized phase II trial of the Minnie Pearl Cancer Research Network. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[27]  B. Bonavida,et al.  Rituximab inhibits p38 MAPK activity in 2F7 B NHL and decreases IL-10 transcription: Pivotal role of p38 MAPK in drug resistance , 2004, Oncogene.

[28]  M. Ghielmini,et al.  Prolonged treatment with rituximab in patients with follicular lymphoma significantly increases event-free survival and response duration compared with the standard weekly x 4 schedule. , 2004, Blood.

[29]  D. Maloney.  Immunotherapy for non-Hodgkin's lymphoma: monoclonal antibodies and vaccines. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[30]  B. Bonavida,et al.  Rituximab-Induced Inhibition of YY1 and Bcl-xL Expression in Ramos Non-Hodgkin’s Lymphoma Cell Line via Inhibition of NF-κB Activity: Role of YY1 and Bcl-xL in Fas Resistance and Chemoresistance, Respectively1 , 2005, The Journal of Immunology.

[31]  N. Mitsiades,et al.  Tumor Cell Expression of CD59 Is Associated With Resistance to CD20 Serotherapy in Patients With B-Cell Malignancies , 2001, Journal of immunotherapy.

[32]  G. Salles,et al.  Second treatment with rituximab in B-cell non-Hodgkin's lymphoma: efficacy and toxicity on 41 patients treated at CHU-Lyon Sud. , 2004, The hematology journal : the official journal of the European Haematology Association.

[33]  G. Salles,et al.  Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor FcgammaRIIIa gene. , 2002, Blood.

[34]  B. Bonavida,et al.  Development of rituximab-resistant lymphoma clones with altered cell signaling and cross-resistance to chemotherapy. , 2007, Cancer research.

[35]  N. Mitsiades,et al.  Tumor Cell Expression of CD59 Is Associated With Resistance to CD20 Serotherapy in Patients With B-Cell Malignancies. , 2001, Journal of immunotherapy : official journal of the Society for Biological Therapy.

[36]  M. Czuczman,et al.  Rituximab anti-CD20 monoclonal antibody therapy in non-Hodgkin's lymphoma: safety and efficacy of re-treatment. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[37]  S. Boehrer,et al.  Anti-CD20 antibody (IDEC-C2B8, rituximab) enhances efficacy of cytotoxic drugs on neoplastic lymphocytes in vitro: role of cytokines, complement, and caspases. , 2002, Haematologica.

[38]  Anton Hagenbeek,et al.  Complement-Induced Cell Death by Rituximab Depends on CD20 Expression Level and Acts Complementary to Antibody-Dependent Cellular Cytotoxicity , 2006, Clinical Cancer Research.

[39]  J. K. Hofmeister,et al.  Clustered CD20 induced apoptosis: src-family kinase, the proximal regulator of tyrosine phosphorylation, calcium influx, and caspase 3-dependent apoptosis. , 2000, Blood cells, molecules & diseases.

[40]  D. R. Anderson,et al.  Targeted anti-cancer therapy using rituximab, a chimaeric anti-CD20 antibody (IDEC-C2B8) in the treatment of non-Hodgkin's B-cell lymphoma. , 1997, Biochemical Society transactions.

[41]  T. Barbui,et al.  CD20 levels determine the in vitro susceptibility to rituximab and complement of B-cell chronic lymphocytic leukemia: further regulation by CD55 and CD59. , 2001, Blood.

[42]  R. Dillman.  Treatment of low-grade B-cell lymphoma with the monoclonal antibody rituximab. , 2003, Seminars in oncology.

[43]  E. Everett,et al.  Reduced expression of CD20 antigen as a characteristic marker for chronic lymphocytic leukemia , 1992, American journal of hematology.

[44]  L. Goodglick,et al.  Inhibition of the Raf–MEK1/2–ERK1/2 Signaling Pathway, Bcl-xL Down-Regulation, and Chemosensitization of Non-Hodgkin’s Lymphoma B Cells by Rituximab , 2004, Cancer Research.

[45]  S. Bernasconi,et al.  Biologic response of B lymphoma cells to anti-CD20 monoclonal antibody rituximab in vitro: CD55 and CD59 regulate complement-mediated cell lysis. , 2000, Blood.

[46]  L. Lanier,et al.  Immune inhibitory receptors. , 2000, Science.

[47]  T. Barbui,et al.  CD 20 levels determine the in vitro susceptibility to rituximab and complement of B-cell chronic lymphocytic leukemia : further regulation by CD 55 and CD 59 , 2001 .

[48]  W. Weng,et al.  Expression of complement inhibitors CD46, CD55, and CD59 on tumor cells does not predict clinical outcome after rituximab treatment in follicular non-Hodgkin lymphoma. , 2001, Blood.

[49]  Signaling events involved in anti-CD20-induced apoptosis of malignant human B cells , 2000, Cancer immunology, immunotherapy : CII.