Targeted Replacement of the NKG2A Checkpoint with CARs at the KLRC1 Locus Enhances NK Cell Immunotherapy in AML
Asunción Borrego Borrego
Centro de Investigación del Cáncer (Universidad de Salamanca-CSIC)
Natural killer (NK) cells represent a promising platform for immunotherapy in acute myeloid leukaemia (AML), where NK cell dysfunction and inhibitory signaling contribute to disease relapse. The inhibitory receptor NKG2A, encoded by KLRC1, is inducibly expressed in response to cytokine stimulation and it has been descrie upon tumour recognition, making it an attractive genomic target for engineering. In this study, we develop a “2-in-1” NK strategy by knocking out KLRC1 and inserting cargo (e.g., CARs or EGFR-based constructs) in-frame into its locus. This approach simultaneously eliminates an inhibitory checkpoint pathway and restricts transgene expression to contexts in which KLRC1 would normally be induced, enabling functional interrogation of NKG2A biology while replacing it with an activating receptor such as a CAR. Targeted integration enhances NK cell anti-leukaemic activity while limiting off-target or tonic activation. In addition, a cis-targeted IL-2 strategy supports selective expansion and persistence of edited NK cells. Together, this dual-editing and selection platform constitutes a next-generation NK cell immunotherapy with improved safety and efficacy for AML.