Spezifische Gene knockouts zur Verbesserung der Proliferation und Persistenz von NK Zellen in der Immuntherapie
Natural killer (NK) cells are potent cytotoxic innate lymphocytes, and the potential of NK cell therapy to treat cancer is currently explored by many clinical trials. However, one of the main drawbacks of such approaches is the limited in vitro expansion capacity of NK cells and, upon transfusion, their insufficient in vivo persistence.
We performed RNA-Seq analysis on NK cells expanded in vitro to determine gene expression changes driven by IL2 and IL15. CRISPR/Cas9-mediated genome editing was used to knockout genes negatively controlling the proliferation and restricting the expansion potential of NK cells.
Primary NK cells were expanded by a factor 500 in vitro using standard pre-clinical protocols. We identified that IL2 and IL15 signaling in these cells involved the induction of negative regulators such as SOCS family genes to limit the proliferation. While meaningful for regular immune reactions, the therapeutic potential of NK cells could be improved by the elimination of inhibitory factors. Thus, we efficiently knocked out CISH, SOCS1 and SOCS2 as shown on the genomic as well as on the protein level. Indeed, these knockout cells had a competitive advantage over unmodified NK cells.
Genome editing of NK cells can be used to increase the proliferative capacity of NK cells. This technical advance will be extended and allow the derivation of NK cells with strongly improved persistence and efficacy in adoptive immunotherapy.
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