The Therapeutic CD38 Monoclonal Antibody Daratumumab Induces Programmed Cell Death via Fcγ Receptor–Mediated Cross-Linking

Emerging evidence suggests that FcγR-mediated cross-linking of tumor-bound mAbs may induce signaling in tumor cells that contributes to their therapeutic activity. In this study, we show that daratumumab (DARA), a therapeutic human CD38 mAb with a broad-spectrum killing activity, is able to induce programmed cell death (PCD) of CD38+ multiple myeloma tumor cell lines when cross-linked in vitro by secondary Abs or via an FcγR. By comparing DARA efficacy in a syngeneic in vivo tumor model using FcRγ-chain knockout or NOTAM mice carrying a signaling-inactive FcRγ-chain, we found that the inhibitory FcγRIIb as well as activating FcγRs induce DARA cross-linking–mediated PCD. In conclusion, our in vitro and in vivo data show that FcγR-mediated cross-linking of DARA induces PCD of CD38-expressing multiple myeloma tumor cells, which potentially contributes to the depth of response observed in DARA-treated patients and the drug’s multifaceted mechanisms of action.

[1]  Bie M. P. Verbist,et al.  Daratumumab depletes CD38+ immune regulatory cells, promotes T-cell expansion, and skews T-cell repertoire in multiple myeloma. , 2016, Blood.

[2]  A. Jakubowiak,et al.  Daratumumab monotherapy in patients with treatment-refractory multiple myeloma (SIRIUS): an open-label, randomised, phase 2 trial , 2016, The Lancet.

[3]  Hao Jiang,et al.  SAR650984 directly induces multiple myeloma cell death via lysosomal-associated and apoptotic pathways, which is further enhanced by pomalidomide , 2016, Leukemia.

[4]  S. Lonial,et al.  Clinical efficacy of daratumumab monotherapy in patients with heavily pretreated relapsed or refractory multiple myeloma. , 2015, Blood.

[5]  A. Palumbo,et al.  Targeting CD38 with Daratumumab Monotherapy in Multiple Myeloma. , 2015, The New England journal of medicine.

[6]  P. Parren,et al.  Antibody-mediated phagocytosis contributes to the anti-tumor activity of the therapeutic antibody daratumumab in lymphoma and multiple myeloma , 2015, mAbs.

[7]  P. Parren,et al.  Direct in Vitro Comparison of Daratumumab with Surrogate Analogs of CD38 Antibodies MOR03087, SAR650984 and Ab79 , 2014 .

[8]  R. Groen,et al.  Modulation of CD38 Expression Levels on Multiple Myeloma Tumor Cells By All-Trans Retinoic Acid Improves the Efficacy of the Anti-CD38 Monoclonal Antibody Daratumumab , 2014 .

[9]  J. Deckert,et al.  SAR650984, A Novel Humanized CD38-Targeting Antibody, Demonstrates Potent Antitumor Activity in Models of Multiple Myeloma and Other CD38+ Hematologic Malignancies , 2014, Clinical Cancer Research.

[10]  A. Mackensen,et al.  A humanized mouse identifies the bone marrow as a niche with low therapeutic IgG activity. , 2014, Cell reports.

[11]  M. Cragg,et al.  Inhibitory FcγRIIb (CD32b) becomes activated by therapeutic mAb in both cis and trans and drives internalization according to antibody specificity. , 2014, Blood.

[12]  J. Ravetch,et al.  Antitumor activities of agonistic anti-TNFR antibodies require differential FcγRIIB coengagement in vivo , 2013, Proceedings of the National Academy of Sciences.

[13]  F. Malavasi,et al.  A CD38/CD203a/CD73 ectoenzymatic pathway independent of CD39 drives a novel adenosinergic loop in human T lymphocytes , 2013, Oncoimmunology.

[14]  M. Cragg,et al.  FcγRΙΙB controls the potency of agonistic anti-TNFR mAbs , 2013, Cancer Immunology, Immunotherapy.

[15]  P. Parren,et al.  Crosstalk between Human IgG Isotypes and Murine Effector Cells , 2012, The Journal of Immunology.

[16]  Y. S. Shin,et al.  An agonistic antibody to human death receptor 4 induces apoptotic cell death in head and neck cancer cells through mitochondrial ROS generation. , 2012, Cancer letters.

[17]  J. Ravetch,et al.  A general requirement for FcγRIIB co-engagement of agonistic anti-TNFR antibodies , 2012, Cell cycle.

[18]  H. Pelicano,et al.  Antibody-induced nonapoptotic cell death in human lymphoma and leukemia cells is mediated through a novel reactive oxygen species-dependent pathway. , 2012, Blood.

[19]  P. Parren,et al.  Exhaustion of Cytotoxic Effector Systems May Limit Monoclonal Antibody-Based Immunotherapy in Cancer Patients , 2012, The Journal of Immunology.

[20]  P. Parren,et al.  Epidermal Growth Factor Receptor (EGFR) Antibody-Induced Antibody-Dependent Cellular Cytotoxicity Plays a Prominent Role in Inhibiting Tumorigenesis, Even of Tumor Cells Insensitive to EGFR Signaling Inhibition , 2011, The Journal of Immunology.

[21]  T. Illidge,et al.  Novel type II anti-CD20 monoclonal antibody (GA101) evokes homotypic adhesion and actin-dependent, lysosome-mediated cell death in B-cell malignancies. , 2011, Blood.

[22]  M. Introna,et al.  Mechanism of Action of Type II, Glycoengineered, Anti-CD20 Monoclonal Antibody GA101 in B-Chronic Lymphocytic Leukemia Whole Blood Assays in Comparison with Rituximab and Alemtuzumab , 2011, The Journal of Immunology.

[23]  Kenneth C. Anderson,et al.  Daratumumab, a Novel Therapeutic Human CD38 Monoclonal Antibody, Induces Killing of Multiple Myeloma and Other Hematological Tumors , 2011, The Journal of Immunology.

[24]  A. Yang,et al.  An Fcγ receptor-dependent mechanism drives antibody-mediated target-receptor signaling in cancer cells. , 2011, Cancer cell.

[25]  C. Klein,et al.  Increasing the efficacy of CD20 antibody therapy through the engineering of a new type II anti-CD20 antibody with enhanced direct and immune effector cell-mediated B-cell cytotoxicity. , 2010, Blood.

[26]  P. Parren,et al.  In vivo cytotoxicity of type I CD20 antibodies critically depends on Fc receptor ITAM signaling. , 2010, Cancer research.

[27]  Michael L. Wang,et al.  Macrophages are an abundant component of myeloma microenvironment and protect myeloma cells from chemotherapy drug-induced apoptosis. , 2009, Blood.

[28]  T. Illidge,et al.  Monoclonal antibodies directed to CD20 and HLA-DR can elicit homotypic adhesion followed by lysosome-mediated cell death in human lymphoma and leukemia cells. , 2009, The Journal of clinical investigation.

[29]  D. Goldenberg,et al.  Induction of apoptosis by cross-linking antibodies bound to human B-lymphoma cells: expression of Annexin V binding sites on the antibody cap. , 2009, Cancer biotherapy & radiopharmaceuticals.

[30]  R. Spaapen,et al.  A bioluminescence imaging based in vivo model for preclinical testing of novel cellular immunotherapy strategies to improve the graft-versus-myeloma effect , 2008, Haematologica.

[31]  C. Watzl,et al.  Serial Killing of Tumor Cells by Human Natural Killer Cells – Enhancement by Therapeutic Antibodies , 2007, PloS one.

[32]  L. Diehl,et al.  Importance of Cellular Microenvironment and Circulatory Dynamics in B Cell Immunotherapy1 , 2005, The Journal of Immunology.

[33]  R. Kyle,et al.  Drug therapy: Multiple myeloma , 2004 .

[34]  S. Knuutila,et al.  Molecular mechanisms of CD99-induced caspase-independent cell death and cell–cell adhesion in Ewing's sarcoma cells: actin and zyxin as key intracellular mediators , 2004, Oncogene.

[35]  G. Tricot,et al.  Flow cytometric immunophenotypic analysis of 306 cases of multiple myeloma. , 2004, American journal of clinical pathology.

[36]  M. Cragg,et al.  CD20-induced lymphoma cell death is independent of both caspases and its redistribution into triton X-100 insoluble membrane rafts. , 2003, Cancer research.

[37]  R. Kimberly,et al.  FcγRs Modulate Cytotoxicity of Anti-Fas Antibodies: Implications for Agonistic Antibody-Based Therapeutics1 , 2003, The Journal of Immunology.

[38]  A. Fischer,et al.  Mechanisms of CD47-induced caspase-independent cell death in normal and leukemic cells: link between phosphatidylserine exposure and cytoskeleton organization. , 2002, Blood.

[39]  V. Fadok,et al.  The phosphatidylserine receptor: a crucial molecular switch? , 2001, Nature Reviews Molecular Cell Biology.

[40]  H. Koeppen,et al.  Isotype-Dependent Inhibition of Tumor Growth In Vivo by Monoclonal Antibodies to Death Receptor 4 , 2001, The Journal of Immunology.

[41]  H. Clevers,et al.  Fluorescence in situ hybridization analysis shows the frequent occurrence of 14q32.3 rearrangements with involvement of immunoglobulin switch regions in myeloma cell lines. , 1999, Cancer genetics and cytogenetics.

[42]  M. V. Vugt,et al.  FcγRIa–γ‐chain complexes trigger antibody‐dependent cell‐mediated cytotoxicity (ADCC) in CD5+ B cell/macrophage IIA1.6 cells , 1998 .

[43]  J. Ledbetter,et al.  Apoptosis of malignant human B cells by ligation of CD20 with monoclonal antibodies. , 1998, Blood.

[44]  E. Vitetta,et al.  Homodimerization of tumor-reactive monoclonal antibodies markedly increases their ability to induce growth arrest or apoptosis of tumor cells. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[45]  G. Kroemer,et al.  Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo , 1995, The Journal of experimental medicine.

[46]  C. Leprince,et al.  B cell antigen receptor-mediated apoptosis. Importance of accessory molecules CD19 and CD22, and of surface IgM cross-linking. , 1995, Journal of immunology.

[47]  J. Ravetch,et al.  FcR γ chain deletion results in pleiotrophic effector cell defects , 1994, Cell.

[48]  H. Asaoku,et al.  Heterogeneous expression of CD32 and CD32-mediated growth suppression in human myeloma cells. , 2006, Haematologica.

[49]  O. Cope,et al.  Multiple myeloma. , 1948, The New England journal of medicine.