Automated manufacturing of clinical-grade BDCA2 CAR NK cells in a closed system for the treatment of blastic plasmacytoid dendritic cell neoplasm.

Recent progress in chimeric antigen receptor (CAR) natural killer (NK) cell therapy has demonstrated their promising potential in cancer immunotherapy. However, most current CAR NK cell manufacturing processes utilize open systems with multiple manual steps, making it challenging to maintain consistent therapeutic quality and regulatory compliance for clinical applications. We specifically developed blood dendritic cell antigen 2 (BDCA2)-targeting CAR NK cells for treating blastic plasmacytoid dendritic cell neoplasm (BPDCN). Here, we present an automated, current good manufacturing practice (cGMP)-compliant Natural Killer Cell Transduction (NKCT) process for producing clinical-grade CAR NK cells on the CliniMACS Prodigy® platform. This closed system integrates cell separation, activation, transduction, expansion, and harvest, thereby reducing contamination risks and ensuring cell product quality. The NKCT process achieved high transduction efficiency using baboon envelope pseudotyped lentiviral vectors (BaEV-LV) produced under cGMP conditions combined with Vectofusin®-1, yielding CAR NK cells with high viability and purity. Both in vitro and in vivo studies demonstrated the potent antitumor activity of CliniMACS Prodigy-manufactured BDCA2 CAR NK cells, highlighting a promising treatment strategy for BPDCN. In summary, this automated NKCT process enables both centralized and decentralized CAR NK manufacturing and facilitates the efficient clinical translation of CAR NK cell therapies.