The anti-CD38 monoclonal antibody SAR650984 (SAR) is showing promising clinical activity in treatment of relapsed and refractory multiple myeloma (MM). Besides effector-mediated antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity, we here define molecular mechanisms of SAR-directed MM cell death and enhanced anti-MM activity triggered by SAR with Pomalidomide (Pom). Without Fc-cross-linking agents or effector cells, SAR specifically induces homotypic aggregation (HA)-associated cell death in MM cells dependent on the level of cell surface CD38 expression, actin cytoskeleton and membrane lipid raft. SAR and its F(ab)’2 fragments trigger caspase 3/7-dependent apoptosis in MM cells highly expressing CD38, even with p53 mutation. Importantly, SAR specifically induces lysosome-dependent cell death (LCD) by enlarging lysosomes and increasing lysosomal membrane permeabilization associated with leakage of cathepsin B and LAMP-1, regardless of the presence of interleukin-6 or bone marrow stromal cells. Conversely, the lysosomal vacuolar H+-ATPase inhibitor blocks SAR-induced LCD. SAR further upregulates reactive oxygen species. Pom enhances SAR-induced direct and indirect killing even in MM cells resistant to Pom/Len. Taken together, SAR is the first therapeutic monoclonal antibody mediating direct cytotoxicity against MM cells via multiple mechanisms of action. Our data show that Pom augments both direct and effector cell-mediated MM cytotoxicity of SAR, providing the framework for combination clinical trials.
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We thank Dr Ti Cai (former employee at Sanofi, currently at EMD Serono, Inc.) for helpful input and all clinical and laboratory members of the Jerome Lipper Multiple Myeloma Center of the Dana-Farber Cancer Institute for support and help for this study. Funding: National Institutes of Health Grants RO1050947, PO1-CA078378 and DF/HCC SPORE in Multiple Myeloma P50CA100707; KCA is an American Cancer Society Clinical Research Professor.
HJ, Y-TT and KCA conceptualized research and formed the hypothesis of this paper; HJ, GA, CA, XF, MZ, LW, ND and LQ designed, performed experiments, collected and analyzed data; HJ and LW performed confocal microscopy; ZS, GY and FA provided transfectants, reagents and analytic tools; NCM, PR and KCA provided MM patient samples; HJ and Y-TT wrote the manuscript; Y-TT and KCA critically evaluated and edited the manuscript.
ZS, GY and FA are employees of Sanofi whose product was used in this research. Y-TT is a consultant for Onyx. PR serves on advisory boards to Millennium, Celgene, Novartis, Johnson & Johnson and Bristol-Myers Squibb. NCM serves on advisory boards to Millennium, Celgene and Novartis. KCA serves on advisory boards to Onyx, Celgene, Gilead, Bristol-Myers Squibb and Sanofi-Aventis and is a scientific founder of Acetylon and Oncopep. The remaining authors declare no competing financial interest.
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Jiang, H., Acharya, C., An, G. et al. SAR650984 directly induces multiple myeloma cell death via lysosomal-associated and apoptotic pathways, which is further enhanced by pomalidomide. Leukemia 30, 399–408 (2016) doi:10.1038/leu.2015.240
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