Multiplex may improve CAR-NK cell potency while raising safety questions about organ toxicity.
Evidence
This commentary summarizes a preclinical CAR-NK cell study using non-viral CAR integration, up to six simultaneous edits, in vitro assays, and xenograft mouse models.
Caveat
Improved potency was accompanied by organ-toxicity signals in mice, so clinical translation remains uncertain.
Simplified
First clinical trials demonstrated the safety of adoptive cell transfer with allogeneic natural killer (NK) cell products from healthy donors, making them an attractive candidate for 'off-the-shelf' chimeric antigen receptor (CAR)-immune cell therapy. However, reduced persistence and inactivation of NK cells by immunosuppressive cues likely limit the performance of CAR-redirected NK cells. Wang and colleagues demonstrate that multiplex allows optimization of the intrinsic functionality of improving their therapeutic potential. In contrast to conventional CRISPR-Cas nucleases, base editing avoided most double-stranded DNA breaks while enabling highly efficient editing at up to six sites simultaneously. The study further demonstrates the feasibility of a non-viral approach to integrate CAR transgene and multiplex base editing of several immune checkpoints in NK cells using a single electroporation. CAR-NK cells harboring up to three base edits demonstrate improved potency over unedited counterparts in vitro. Xenograft mouse models confirmed increased potency, but also indicated signs of organ toxicity - a phenomenon that will require future studies prior to clinical translation. The study demonstrates that CRISPR base editing is a powerful tool to unleash the full cytotoxic potential of NK cells, but it also warrants the question: How many internal breaks can be removed without hurting CAR-NK cell therapy's impeccable safety record?
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Competing interests: TB and DLW have no competing interests with regard to this work. TB is an inventor on patent applications related to gene editing and cell therapies, including non-viral CAR-NK cell approaches. TB was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - 493624332, INDEEP Clinician Scientist Program, and by the MSNZ Frankfurt (German Cancer Aid, Funding ID: 70113301). DLW is an inventor on several patent applications related to gene editing and cell therapies, including CAR-NK cell approaches, and is a co-founder of TCBalance Biopharmaceuticals GmbH.
