Nanobody-based CD38-specific heavy chain antibodies induce killing of multiple myeloma and other hematological malignancies

Levin Schriewer^1,2*, Kerstin Schütze^1,2*, Katharina Petry^1,2*, Julia Hambach^1,2, William Fumey^1,2, Julia Koenigsdorf^1,2, Natalie Baum^1,2, Stephan Menzel², Björn Rissiek³, Kristoffer Riecken^4,6, Boris Fehse^4,6, Jana Larissa Röckendorf^1,2, Joanna Schmid^1,2, Birte Albrecht^1,2, Hans Pinnschmidt⁵, Francis Ayuk⁶, Nicolaus Kröger⁶, Mascha Binder^7,8, Gunter Schuch⁹, Timon Hansen⁹, Friedrich Haag², Gerhard Adam¹, Friedrich Koch-Nolte^2#, Peter Bannas^1#✉

1. Department of Diagnostic and Interventional Radiology and Nuclear medicine, University Medical Center, Hamburg-Eppendorf, Germany.
2. Institute of Immunology, University Medical Center, Hamburg-Eppendorf, Germany.
3. Department of Neurology, University Medical Center, Hamburg-Eppendorf, Germany.
4. Research Department Cell and Gene Therapy, University Medical Center, Hamburg-Eppendorf, Germany.
5. Institute of Medical Biometry and Epidemiology, University Medical Center, Hamburg-Eppendorf, Germany.
6. Department of Stem Cell Transplantation, University Medical Center, Hamburg-Eppendorf, Germany.
7. Department of Oncology and Hematology, University Medical Center, Hamburg-Eppendorf, Germany.
8. Department of Haematology and Oncology, University Hospital Halle, Halle, Germany.
9. Hematology and Oncology Center Altona (HOPA), Hamburg, Germany.
Nanobody® is a trademark of Ablynx. In this paper we use nanobody as the generic term for the recombinant VHH domain of a llama heavy chain antibody.
* LS, KS, and KP contributed equally
# FK-N and PB share senior authorship

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Citation:
Schriewer L, Schütze K, Petry K, Hambach J, Fumey W, Koenigsdorf J, Baum N, Menzel S, Rissiek B, Riecken K, Fehse B, Röckendorf JL, Schmid J, Albrecht B, Pinnschmidt H, Ayuk F, Kröger N, Binder M, Schuch G, Hansen T, Haag F, Adam G, Koch-Nolte F, Bannas P. Nanobody-based CD38-specific heavy chain antibodies induce killing of multiple myeloma and other hematological malignancies. Theranostics 2020; 10(6):2645-2658. doi:10.7150/thno.38533. Available from http://www.thno.org/v10p2645.htm

Abstract

Rationale: CD38 is a target for the therapy of multiple myeloma (MM) with monoclonal antibodies such as daratumumab and isatuximab. Since MM patients exhibit a high rate of relapse, the development of new biologics targeting alternative CD38 epitopes is desirable. The discovery of single-domain antibodies (nanobodies) has opened the way for a new generation of antitumor therapeutics. We report the generation of nanobody-based humanized IgG1 heavy chain antibodies (hcAbs) with a high specificity and affinity that recognize three different and non-overlapping epitopes of CD38 and compare their cytotoxicity against CD38-expressing hematological cancer cells in vitro, ex vivo and in vivo.

Methods: We generated three humanized hcAbs (WF211-hcAb, MU1067-hcAb, JK36-hcAb) that recognize three different non-overlapping epitopes (E1, E2, E3) of CD38 by fusion of llama-derived nanobodies to the hinge- and Fc-domains of human IgG1. WF211-hcAb shares the binding epitope E1 with daratumumab. We compared the capacity of these CD38-specific hcAbs and daratumumab to induce CDC and ADCC in CD38-expressing tumor cell lines in vitro and in patient MM cells ex vivo as well as effects on xenograft tumor growth and survival in vivo.

Results: CD38-specific heavy chain antibodies (WF211-hcAb, MU1067-hcAb, JK36-hcAb) potently induced antibody-dependent cellular cytotoxicity (ADCC) in CD38-expressing tumor cell lines and in primary patient MM cells, but only little if any complement-dependent cytotoxicity (CDC). In vivo, CD38-specific heavy chain antibodies significantly reduced the growth of systemic lymphomas and prolonged survival of tumor bearing SCID mice.

Conclusions: CD38-specific nanobody-based humanized IgG1 heavy chain antibodies mediate cytotoxicity against CD38-expressing hematological cancer cells in vitro, ex vivo and in vivo. These promising results of our study indicate that CD38-specific hcAbs warrant further clinical development as therapeutics for multiple myeloma and other hematological malignancies.