ErbB2/HER2-targeted CAR-NK cells eliminate breast cancer cells in an organoid model that recapitulates tumor progression

Chimeric antigen receptor-engineered NK cells hold promise for adoptive cancer immunotherapy. In one such approach, the ErbB2 (HER2)-specific CAR-NK cell line NK-92/5.28.z is under investigation as an off-the-shelf therapy in a phase I trial in glioblastoma patients. To evaluate activity of NK-92/5.28.z cells against ErbB2-positive breast cancer, here we developed an organoid model derived from CKP mice that allows conditional activation of oncogenic driver mutations. Expression of ErbB2 and Cre recombinase in CKP mammary epithelial cells induced malignant transformation, with the resulting EC-CKP cells characterized by neoplastic morphology, loss of p53, and constitutive activation of the MAP kinase pathway. NK-92/5.28.z cells demonstrated potent CAR-mediated cytotoxicity against EC-CKP organoids, with tumor cell lysis dependent on exposure time and organoid size. In vivo passaging of EC-CKP organoids revealed cellular plasticity and induced an EMT phenotype associated with increased resistance to standard therapies. Importantly, NK-92/5.28.z cells retained high and specific cytotoxicity against these breast cancer cells in vitro and in an aggressive organoid-based in vivo mouse model that reflects advanced-stage disease. Our data highlight the therapeutic potential of NK-92/5.28.z cells against ErbB2-positive breast cancer, supporting their further development toward clinical application.