Scalable Generation of Hematopoietic Stem Cells Engineered Off-the-Shelf Mono-Specific Cytotoxic T Cells Targeting Solid Tumors

However, its clinical application remains constrained by major limitations, including manufacturing complexity in autologous settings and risks of graft-versus-host disease (GvHD), HLA restriction and donor variability in allogeneic contexts. To address these challenges, we present a scalable platform for the generation of hematopoietic stem and progenitor cell (HSPC)-derived, mono-specific cytotoxic T cells targeting the NY-ESO-1 antigen.

Using an ex vivo, feeder-free, serum-free culture system, we differentiate gene-engineered HSPCs into allogeneic T cells (AlloESO-T) with a uniform cytotoxic phenotype and robust antitumor activity. These cells employ dual tumor-targeting strategies by antigen-specific killing via a transgenic TCR, and antigen-independent cytotoxicity via natural killer receptors. Compared to conventional PBMC-derived TCR-T cells, Allogeneic ESO-T cells display superior cytotoxic potency, selective solid tumor homing, durable killing persistence, and resilience against immune evasion.