Abstract 2970: Comparison of anti-CD38 antibodies in vitro mechanisms of action in multiple myeloma
Authors: Kinder Michelle , Ta Mi , Axel Amy , Wong Amy , Rudnick Stephen , Goeij Bart de , Bueren Jeroen Lammerts van , Babich Alex , Mendonca Mark , Sendecki Jocelyn , Chiu Christopher , Bellew Kevin , Kane Colleen 
Published: August 2019

Abstract
Background: The outcome of multiple myeloma (MM) patients has been dramatically improved by the monoclonal antibody (mAb) daratumumab (DARA). DARA is a human IgG1κ mAb specific for CD38 and approved for the treatment of newly diagnosed and relapsed/refractory MM as a monotherapy and in combination with standard of care. Isatuximab and TAK-079 are additional CD38 targeting mAbs in clinical development by Sanofi and Takeda, respectively. These mAbs are reported to bind different epitopes and induce lysis of MM tumor cells via multiple mechanisms. It is unclear how the pleiotropic mechanisms collectively impact tumor cytolysis and exhibit anti-tumor effects in a comprehensive ex vivo immune milieu.
Methods: For research purposes, DARA was compared with representative mAbs based on the published patent sequences for isatuximab and TAK-079, these surrogate analogs will hereafter be referred to as Sanofi anti-CD38 and Takeda anti-CD38. Mechanism of action (MOA) studies were done to compare antibody dependent cell-mediated cytotoxicity (ADCC), antibody dependent cell phagocytosis (ADCP), complement mediated cytotoxicity (CDC), and early and late detection of apoptosis. In addition, fresh whole blood from healthy donors was used to assess the cumulative effect of the MOAs on MM cell lines.
Results: We tested the CDC activity of the anti-CD38s on multiple MM cell lines with a range of CD38 surface expression and CDC sensitivity levels. In LP-1 and MOLP-8 MM cell lines, DARA resulted in higher levels CDC activity as compared to the other anti-CD38 mAbs. In ADCC and ADCP assays, all 3 anti-CD38 mAbs induced similar levels of MM cell death and phagocytosis. Apoptosis was also assessed in the presence and absence of FcR crosslinking. The Annexin V assay replicated published results that only the Sanofi anti-CD38 was able to induce phosphatidylserine translocation to the cell surface without FcR crosslinking. However, in a robust 5-day cytotoxicity assay detecting metabolically active cells, all 3 antibodies elicited comparable high levels of cell death in the presence of the FcR crosslinker and low levels in its absence. We also performed assays with fresh whole blood from healthy volunteers (n=6) and determined the cumulative effect of the anti-CD38 mAbs on LP-1 and MOLP-8 MM cell lines. DARA demonstrated a higher maximal cytotoxicity than the other mAbs. Moreover, DARA had a significantly lower EC50 than Takeda anti-CD38 in both cell lines and lower than Sanofi-anti-CD38 in MOLP-8.
Conclusion: DARA and surrogate analogs of Sanofi anti-CD38 and Takeda anti-CD38 have similar MOAs with the exception of higher CDC demonstrated by DARA which may contribute to the observed improved efficacy of DARA in a comprehensive immune milieu in vitro. As DARA is the only approved CD38 mAb, it remains to be determined in clinical trials if these in vitro differences lead to differences in clinical benefit.
Citation Format: Michelle Kinder, Mi Ta, Amy Axel, Amy Wong, Stephen Rudnick, Bart de Goeij, Jeroen Lammerts van Bueren, Alex Babich, Mark Mendonca, Jocelyn Sendecki, Christopher Chiu, Kevin Bellew, Colleen Kane. Comparison of anti-CD38 antibodies in vitro mechanisms of action in multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2970.