Published OnlineFirst January 22, 2020; DOI: 10.1158/1078-0432.CCR-19-2299 CLINICAL CANCER RESEARCH | TRANSLATIONAL CANCER MECHANISMS AND THERAPY Preclinical Activity of JNJ-7957, a Novel BCMAÂCD3 Bispecific Antibody for the Treatment of Multiple Myeloma, Is Potentiated by Daratumumab Kristine A. Frerichs1, Marloes E.C. Broekmans1, Jhon A. Marin Soto1, Berris van Kessel1, Martijn W. Heymans2, Lisa C. Holthof1, Christie P.M. Verkleij1, Rengasamy Boominathan3, Bhavesh Vaidya3, Jocelyn Sendecki3, Amy Axel3, Francois Gaudet3, Kodandaram Pillarisetti3, Sonja Zweegman1, Homer C. Adams III3, Tuna Mutis1, and Niels W.C.J. van de Donk1 ABSTRACT ◥ Purpose: Multiple myeloma (MM) patients with disease refrac- expression did not affect activity of JNJ-7957. High T-cell tory to all available drugs have a poor outcome, indicating the need frequencies and high effector:target ratios were associated with for new agents with novel mechanisms of action. improved JNJ-7957–mediated lysis of MM cells. The PD-1/ Experimental design: We evaluated the anti-MM activity of the PD-L1 axis had a modest negative impact on JNJ-7957 activity fully human BCMAÂCD3 bispecific antibody JNJ-7957 in cell lines against tumor cells from daratumumab-na€ve MM patients. and bone marrow (BM) samples. The impact of several tumor- and Soluble BCMA impaired the ability of JNJ-7957 to kill MM host-related factors on sensitivity to JNJ-7957 therapy was also cells, although higher concentrations were able to overcome evaluated. this negative effect. Results: We show that JNJ-7957 has potent activity against Conclusions: JNJ-7957 effectively kills MM cells ex vivo, 4 MM cell lines, against tumor cells in 48 of 49 BM samples including those from heavily pretreated MM patients, whereby obtained from MM patients, and in 5 of 6 BM samples several components of the immunosuppressive BM microenvi- obtained from primary plasma cell leukemia patients. JNJ- ronment had only modest effects on its killing capacity. Our 7957 activity was significantly enhanced in patients with prior findings support the ongoing trial with JNJ-7957 as single agent daratumumab treatment, which was partially due to enhanced and provide the preclinical rationale for evaluating JNJ-7957 in killing capacity of daratumumab-exposed effector cells. BCMA combination with daratumumab in MM. Introduction small subset of normal mature B cells, as well as normal plasma cells and MM cells. BCMA is involved in differentiation of normal B In the last decade, survival of multiple myeloma (MM) patients has cells to plasma cells (3, 4). In addition, binding of a proliferation- markedly improved due to the introduction of immunomodulatory inducing ligand (APRIL) or B-cell activating factor (BAFF), which drugs (IMiD), proteasome inhibitors (PI), and the CD38-targeting has 1,000-fold lower affinity than APRIL, to BCMA promotes MM antibody daratumumab. However, patients with disease refractory to growth and induces immunosuppression in the bone marrow (BM) these agents have a very poor outcome (1). Also, newly diagnosed MM microenvironment (5–7). The selective expression of BCMA on patients with high-risk cytogenetic abnormalities have a reduced normal and malignant plasmablasts and plasma cells renders it an survival, when compared with patients with standard-risk disease (2). attractive target for novel MM treatment strategies (8, 9). Indeed, This indicates that there is still a need for new agents with novel BCMA antibody–drug conjugates and BCMA chimeric antigen recep- mechanisms of action. tor (CAR) T cells show much promise in patients with advanced B-cell maturation antigen (BCMA) is a cell membrane–bound MM (10–14). Furthermore, preclinical studies have demonstrated that tumor necrosis factor receptor family member and expressed on a BCMAÂCD3 bispecific antibodies and bispecific T-cell engagers (BiTE) have marked anti-MM activity (9, 15, 16). In addition, pre- liminary results from a phase I study show encouraging single-agent 1Amsterdam UMC, Vrije Universiteit Amsterdam, Department of Hematology, fi 2 activity of AMG420, a BCMA-targeting BiTE (17). These bispeci c Cancer Center Amsterdam, Amsterdam, the Netherlands. Amsterdam UMC, antibodies and BiTEs redirect T cells to the BCMA-positive tumor cell, Vrije Universiteit Amsterdam, Department of Epidemiology and Biostatistics, Amsterdam, the Netherlands. 3Janssen Research and Development, LLC, Spring which is followed by perforin and granzyme release, and ultimately House, Pennsylvania. MM cell death. Killing of tumor cells by redirected T cells is inde- pendent of T-cell receptor specificity. However, MM patients with Note: Supplementary data for this article are available at Clinical Cancer Research Online (http://clincancerres.aacrjournals.org/). advanced disease have a more immunosuppressive BM microenvi- ronment, including increased regulatory T-cell (Treg) counts, as well Prior presentation: Presented at the 23rd annual meeting of the European Hematology Association, Stockholm, 17 June 2018 (oral presentation). as upregulation of inhibitory immune checkpoints such as pro- grammed death ligand-1 (PD-L1) on MM cells and programmed cell Corresponding Author: Niels W.C.J. van de Donk, Vrije Universiteit Medical death-1 (PD-1) on T cells, which may limit the activity of bispecific Center, de Boelelaan 1117, Amsterdam 1081 HV, the Netherlands. Phone: 31-20- 4442604; Fax: 31-20-44442601; E-mail: [email protected] antibodies in these patients (18, 19). In addition, prior immunosup- pressive anti-MM therapies may hamper T-cell function and affect the Clin Cancer Res 2020;XX:XX–XX activity of these T-cell–redirecting therapies (20). doi: 10.1158/1078-0432.CCR-19-2299 Interestingly, next to its classic Fc-dependent immune effector Ó2020 American Association for Cancer Research. functions, daratumumab also has immunomodulatory effects through AACRJournals.org | OF1 Downloaded from clincancerres.aacrjournals.org on September 25, 2021. © 2020 American Association for Cancer Research. Published OnlineFirst January 22, 2020; DOI: 10.1158/1078-0432.CCR-19-2299 Frerichs et al. therapy was given according to the approved dose and schedule Translational Relevance (16 mg/kg weekly for 8 weeks, then every 2 weeks for 16 weeks, and There is a need for new agents with novel mechanisms of action every 4 weeks until progression). in multiple myeloma (MM). B-cell maturation antigen (BCMA) The study site ethics committee approved the protocols, which has been identified as a novel target for MM treatment, due to its were conducted according to the principles of the Declaration of restricted expression on plasma cells. We show that the novel Helsinki, the International Conference on Harmonization, and the BCMAÂCD3 bispecific antibody, JNJ-7957, induces potent T-cell– Guidelines for Good Clinical Practice. All patients gave written mediated killing of MM cell lines. JNJ-7957 also effectively elim- informed consent. inated primary MM cells in 48 of 49 bone marrow (BM) samples obtained from both newly diagnosed and heavily pretreated Antibodies and reagents MM patients. Interestingly, JNJ-7957 activity was significantly JNJ-7957 (JNJ-64007957) and daratumumab were provided by enhanced in patients with prior daratumumab treatment, which Janssen Pharmaceuticals. CNTO7008 (CD3 Â null), BC3B4 (BCMA Â was partially due to enhanced killing capacity of daratumumab- null), and CNTO3930 (IgG isotype control) were used as control exposed effector cells. The immunosuppressive BM microenviron- antibodies and provided by Janssen Pharmaceuticals. Recombinant ment had only limited inhibitory effects on JNJ-7957 activity. human BCMA and recombinant human APRIL were obtained Altogether, these data strengthen the preclinical rationale for the from Sigma-Aldrich. Blinatumomab (Amgen) was obtained via the ongoing clinical trial evaluating JNJ-7957. In addition, our data clinical pharmacy of Amsterdam University Medical Center. indicate that clinical evaluation of the combination of daratumu- mab and JNJ-7957 is warranted. Flow-cytometric analysis of BM and blood samples from MM patients BM-localized MM cells were identified and analyzed for cell- surface marker expression levels by staining 1.0 Â 106 cells/mL with elimination of immune suppressor cells, induction of T-cell expansion, CD138 PE, CD56 PC7, CD45 Krome Orange (all Beckman Coulter), and enhancement of the cytolytic capacity of T cells (21–26), which CD269 (BCMA) APC (BioLegend), CD274 (PD-L1) BV421, and may potentially improve the efficacy of T-cell–redirecting agents to kill CD19 APC-H7 (both Becton Dickinson). To assess CD38 expres- MM cells. sion irrespective of ongoing or recent daratumumab treatment, We therefore evaluated in this study the preclinical activity of the cells were also stained with HuMax-003 FITC, which binds to a novel fully human BCMAÂCD3 bispecific antibody (JNJ-7957), CD38 epitope distinct from the epitope bound by daratumumab which was developed using the Genmab DuoBody technology (Genmab/Janssen Pharmaceuticals). and has silenced Fc function to eliminate Fc-dependent immune Immune cell subsets in whole PB or BM aspirates were identified effector functions (27). The antitumor efficacy was tested in MM and analyzed for cell-surface marker expression levels by staining cell lines and in BM samples obtained from patients with newly 1.0 Â 106 cells/mLwithCD45KromeOrange,CD56PC7(both diagnosed (ND) or relapsed/refractory (RR) MM, including heavily Beckman Coulter), CD14 APC-H7, CD19 APC-H7, CD3 V450, pretreated patients with disease refractory to IMiDs, PIs, and CD38 CD4 APC-H7 or PE, CD8 FITC, CD45-RA APC, CD127 PE.Cy7, antibodies. We analyzed the impact of several host and tumor CD62L PE, CD274 (PD-1) BV421, CD16 APC, HLA-DR APC-H7 characteristics on ex vivo JNJ-7957 response. Furthermore, we (all Becton Dickinson), CD25 PE (Dako), and CD38 HuMax-003 þ þ evaluated the potential ability of daratumumab to improve the FITC. Na€ve T cells were defined as CD45RA CD62L , effector À À efficacy of JNJ-7957 to kill MM cells by virtue of its immunomod- memory (EM) T cells as CD45RA CD62L , central memory (CM) À þ ulatory effects. T cells as CD45RA CD62L , and terminally differentiated EM T þ À cells expressing CD45RA (TEMRA) as CD45RA CD62L (28).