Nanobody‐Engineered Natural Killer Cell Conjugates for Solid Tumor Adoptive Immunotherapy

Cancer immunotherapy based on natural killer (NK) cells is demonstrated to be a promising strategy. However, NK cells are deficient in ligands that target specific tumors, resulting in limited antitumor efficacy. Here, a glycoengineering approach to imitate the chimeric antigen receptor strategy and decorate NK cells with nanobodies to promote NK‐based immunotherapy in solid tumors is proposed. Nanobody 7D12, which specifically recognizes the human epidermal growth factor receptor (EGFR) that is overexpressed on many solid tumors, is coupled to the chemically synthesized DBCO‐PEG4‐GGG‐NH2 by sortase A‐mediated ligation to generate DBCO‐7D12. The NK92MI cells bearing azide groups are then equipped with DBCO‐7D12 via bioorthogonal click chemistry. The resultant 7D12‐NK92MI cells exhibit high specificity and affinity for EGFR‐overexpressing tumor cells in vitro and in vivo by the 7D12‐EGFR interaction, causing increased cytokine secretion to more effectively kill EGFR‐positive tumor cells, but not EGFR‐negative cancer cells. Importantly, the 7D12‐NK92MI cells also show a wide anticancer spectrum and extensive tumor penetration. Furthermore, mouse experiments reveal that 7D12‐NK92MI treatment achieves excellent therapeutic efficacy and outstanding safety. The authors’ works provide a cell modification strategy using specific protein ligands without genetic manipulation and present a potential novel method for cancer‐targeted immunotherapy by NK cells. A nanobody‐NK cell conjugate is developed using a non‐chimeric antigen receptor (non‐CAR) technique. The 7D12‐engineered NK cells can actively target epidermal growth factor receptor (EGFR)‐positive cancer cells and display remarkably enhanced antitumor immunity toward several types of cancer cells in vitro. The nanobody‐NK cell conjugates gain excellent cancer tissue penetration capability and exhibit improved therapeutic efficacy in vivo.