Challenges in αCD38-chimeric antigen receptor (CAR)-expressing natural killer (NK) cell-based immunotherapy in multiple myeloma: Harnessing the CD38dim phenotype of cytokine-stimulated NK cells as a strategy to prevent fratricide

Background aims: Adoptive cell therapy with chimeric antigen receptor (CAR)-expressing natural killer (NK) cells is an emerging approach that holds promise in multiple myeloma (MM). However, the generation of CAR-NK cells targeting CD38 is met with obstacles due to the expression of CD38 on NK cells. Knock-out of CD38 is currently explored as a strategy, although the consequences of the lack of CD38 expression with regards to engraftment and activity in the bone marrow microenvironment are not fully elucidated. Here, we present an alternative approach by harnessing the CD38dim phenotype occurring during long-term cytokine stimulation of primary NK cells.Methods: Primary NK cells were expanded from peripheral blood mononuclear cells by long-term IL-2 stimu-lation. During expansion, the CD38 expression was monitored in order to identify a time point when intro-duction of a novel affinity-optimized aCD38-CAR confered optimal viability, i.e. prevented fratricide. CD38dim NK cells were trasduced with retroviral vectors encoding for the CAR trasngene and their functional-ity was assessed in in vitro activation and cytotoxicity assays.Results: We verified the functionality of the aCD38-CAR-NK cells against CD38+ cell lines and primary MM cells. Importantly, we demonstrated that aCD38-CAR-NK cells derived from patients with MM have increased activity against autologous MM samples ex vivo. Conclusions: Overall, our results highlight that incorporation of a functional aCD38-CAR construct into a suit-able NK-cell expansion and activation protocol results in a potent and feasible immunotherapeutic strategy for the treatment of patients with MM. & COPY; 2023 International Society for Cell & Gene Therapy. Published by Elsevier Inc.This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)