Daratumumab, a novel human CD38 monoclonal antibody for the treatment of B cell Non Hodgkin Lymphoma

Transcription

1 Daratumumab, a novel human CD38 monoclonal antibody for the treatment of B cell Non Hodgkin Lymphoma Anna Vidal Department of Hemato Oncology IDIBAPS, Barcelona, Spain

2 B cell lymphoid malignancies seen through the lymph node Bone marrow Lymph Node Plasma cell Precursor B cell Naive B cell Mantle Germinal center Marginal zone Multiple Myeloma Lymphoplasmacytic lymphoma Leukemia/ lymphoma lymphoblastic/o B Chronic lymphocytic leukemia del 13q Del 11q Del 17p NOTCH1 SF3B Mantle Cell lymphoma t(11;14) Cyclin D1 P53 ATM P16 BMI Follicular lymphoma t(14; 18) BCL2 Burkitt lymphoma Diffuse Large B cell Lymphoma MALT lymphoma splenic/marginal zone lymphoma Memory B cell Chronic lymphocytic leukemia Precursor B cell malignancies Peripheral B cell malignancies (mature)

3 Biology of CD38 Malavasi, BLOOD 211 CD38 appears to be a global molecular bridge to the environment, promoting survival/proliferation over apoptosis of B cells on their way to and after neoplastic transformation

4 Daratumumab (DARA): Human CD38 mab In development for Multiple Myeloma (MM) Induces potent ADCC (Antibody dependent Cellular Cytotoxicity) in Burkitt lymphoma and MM derived cell lines as well as in patient MM cells (both with autologous and allogeneic effector cells). Strong ability to induce CDC (Complement dependent Cytotoxicity) in patient MM cells. Under evaluation in phase I/II clinical trials in patients with MM Our primary goal is to evaluate the activity of daratumumab against CD38 expressing tumor cells in B cell lymphomas: Mantle cell lymphoma (MCL) Chronic lymphocytic leukemia (CLL) Follicular lympoma (FL) Heterogeneous CD38 levels High CD38 levels

5 Aims 1. To understand the mechanisms of DARA induced cytotoxicity in MCL, FL and CLL cell lines and primary cells. 2. To evaluate the effect of DARA in combination with lenalidomide on ADCC in CLL cells. 3. To analyze the effect of DARA in SDF1 /CXCL12 induced migration in vitro and in vivo homing in CLL models. 4. To explore in vivo antitumoral effect of DARA in MCL, FL and CLL xenograft mouse models.

6 1. Mechanisms of DARA induced cytotoxicity in MCL, FL and CLL cell lines and primary cells: ADCC PBMCs from healthy donors Ratio T:E = 1:5, 4h treatment Calcein release assay NK cells % Lysis Cell lines DARA g/ml HBL2 REC JEKO WSU-FSCCL RL SC1 DOHH2 MEC2 JVM13 DAUDI MCL FL CLL BL (positive control) Monocytes /macrophages % Lysis Primary material MCL2 FL2 CLL DARA g/ml Daratumumab induces ADCC in the presence of external effectors in MCL, FL and CLL

7 2. Effect of lenalidomide on DARA Induced ADCC in primary CLL cells CLL3 * p<.5 * * CLL1 * p<.5 * * % Lysis 4 2 % Lysis antibody concentration g/ml antibody concentration g/ml IgG1 IgG1+LENA DARA DARA+LENA ** p<.1 ** p<.1 PBMCs from healthy donors (pretreated or not with Lena 3uM 72h) Calcein release assay Lenalidomide pretreatment of PBMCs significantly increases Daratumumab ADCC activity

8 3. Effect of DARA in SDF1 /CXCL12 induced migration in vitro and in vivo homing in CLL In vitro * *p<.1 Total number of migrated cells CLL1 CLL3 CLL4 CLL5 CLL8 **p<.1 Isotype ctrl DARA anti-cxcr4 Isotype ctrl DARA anti-cxcr4 SDF1 +SDF1

9 In vivo Day Day 1 Day 2 NSG mice Mice Ab Pretreatment (isotype ctrl, Dara, Anti CXCR4) Fresh CLL iv Injection of 2x1 6 Cell isolation from PB, BM and Spleen Cell count by flow cytometry (CD45+/CD19+/CD5+ cells) spleen Isotype ctrl spleen **p<.1 * p<.1 SSC CD45 Pacific Blue CD5 PeCy5 total number of cells DARA SSC FSC FSC Anti CXCR4 CD45 Pacific Blue FSC FSC CD5 PeCy5 CD19 FITC CD19 FITC Isotype ctrl DARA anti-cxcr4 SSC CD45 Pacific Blue CD5 PeCy5 FSC FSC CD19 FITC Daratumumab interferes with in vitro CLL migration and in vivo homing to spleen

10 4. In vivo antitumoral effect of DARA in MCL, FL and CLL xenograft mouse models FL (RL cell line) Day 1 Day 7 Day 14 Day 21 Day 28 Day 32 SCID mice Tumor Volume (mm 3 ) Isotype ctrl DARA Ab treatment n= 5 ** p<.1 IgG1 DARA time (days) RFU (relative fluorescence units) 6.3x x x x1 6 IRDye8 probe (LI COR, Odyssey)

11 MCL (REC cell line) Day 1 Day 8 Day 14 Day 21 Day 28 Day 31 SCID mice 8 Tumor Volume (mm3) Isotype ctrl n= 6 DARA Ab treatment *** p< time (days) Tumor weight (g) Spleen weight (g) 6 4 2,15,1,5, *** p<.1 IgG1 B12 Daratumumab *** p<.1 IgG1 B12 Daratumumab Daratumumab inhibits tumor growth in FL and MCL mouse models

12 CLL (MEC 2) Day Day 1 Day 7 Day 14 Day 21 Day 28 Day Day 9 SCID mice Ab treatment Daratumumab increases overall survival in a systemic CLL mouse model

13 Conclusions DARA induces ADCC in MCL, FL and CLL primary cells and cell lines in the presence of PBMC effector cells. Lenalidomide pretreatment of effector PBMCs significantly enhances ADCC induced by DARA in CLL primary cells. DARA significantly reduces SDF1 /CXCL12 induced migration in vitro and in vivo homing to spleen of CLL cells. In vivo, DARA inhibits tumor growth in MCL and FL sc mouse xenograft models and improves overall survival of a CLL systemic mouse model.

14 Thanks for your attention! Alba Matas Céspedes Vanina Rodriguez Patricia Pérez Galán Gael Roué Armando López-Guillermo Dolors Colomer Elias Campo Jeroen J. Lammerts van Bueren Paul W.H.I. Parren Joost M. Bakker Wim Bleeker Parul Doshi Adrian Wiestner

15 Daratumumab does not induce CDC in MCL, FL or CLL cells: possible explanations

16 Chronic lymphocytic leukemia (CLL) Models of study & CD38 immunotherapy Limited efficacy of anti CD2 antibodies High CD38 marker ofbad prognosis and disease progression CD38 is upregulated in the proliferating fraction (Damle RN, Blood 27) CD38 identifies CLL with high migratory potential (Deaglio S, Blood 27) CD38 cooperates with CXCR4 induced migration and in sustaining BCR mediated signals (Vaisitti et al, Leukemia 21) CD38 + CLL cell CD38 CLL cell Mantle cell lymphoma (MCL) Blastoid variant shows higher CD38 expression Follicular lymphoma (FL) Bortezomib resistant MCL cells express higher CD38 levels (Pérez Galán, P. Blood 211) Possible role in migration and microenvironment interactions as found in CLL Invariable high CD38 expression (GC cell) Alternative to anti CD2 therapies in relapse setting Possible role in migration and microenvironment interactions as found in CLL