Rituximab (RTX) plus chemotherapy (R-CHOP) applied as a first-line therapy for lymphoma leads to a relapse in approximately 40% of the patients. Therefore, novel approaches to treat aggressive lymphomas are being intensively investigated. Several RTX-resistant (RR) cell lines have been established as surrogate models to study resistance to R-CHOP. Our study reveals that RR cells are characterized by a major downregulation of CD37, a molecule currently explored as a target for immunotherapy. Using CD20 knockout (KO) cell lines, we demonstrate that CD20 and CD37 form a complex, and hypothesize that the presence of CD20 stabilizes CD37 in the cell membrane. Consequently, we observe a diminished cytotoxicity of anti-CD37 monoclonal antibody (mAb) in complement-dependent cytotoxicity in both RR and CD20 KO cells that can be partially restored upon lysosome inhibition. On the other hand, the internalization rate of anti-CD37 mAb in CD20 KO cells is increased when compared to controls, suggesting unhampered efficacy of antibody drug conjugates (ADCs). Importantly, even a major downregulation in CD37 levels does not hamper the efficacy of CD37-directed chimeric antigen receptor (CAR) T cells. In summary, we present here a novel mechanism of CD37 regulation with further implications for the use of anti-CD37 immunotherapies.

Biobank Maria Sklodowska Curie National Research Institute of Oncology Warsaw Poland

Central European Institute of Technology Masaryk University Brno Czech Republic

Department of Biology Faculty of Medicine Masaryk University Brno Czech Republic

Department of General Oncological and Functional Urology Medical University of Warsaw Warsaw Poland

Department of Hematology Institute of Hematology and Transfusion Medicine Warsaw Poland

Department of Immunology Medical University of Warsaw Warsaw Poland

Department of Pathology Maria Sklodowska Curie National Research Institute of Oncology Warsaw Poland

Division of Antibody Based Immunotherapy Department of Medicine 2 Kiel University and University Medical Center Schleswig Holstein Kiel Germany

Doctoral School Medical University of Warsaw Warsaw Poland

Doctoral School of Translational Medicine Mossakowski Medical Research Institute Polish Academy of Sciences Warsaw Poland

Faculty of Chemistry Wroclaw University of Science and Technology Wroclaw Poland

Faculty of Medicine Lazarski University Warsaw Poland

Faculty of Medicine Wroclaw University of Science and Technology Wroclaw Poland

Laboratory of Immunology Mossakowski Medical Research Institute Polish Academy of Sciences Warsaw Poland

Translational Research Unit Department of Cellular Therapy Department of Oncology Oslo University Hospital Oslo Norway

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