Tcf-1 protects anti-tumor TCR-engineered CD8(+) T-cells from GzmB mediated self-destruction

BACKGROUND: T-cell longevity is undermined by antigen-driven differentiation programs that render cells prone to attrition through several mechanisms. CD8 (+ )T cells that express the Tcf-1 transcription factor have undergone limited differentiation and exhibit stem-cell-like replenishment functions that facilitate persistence. We engineered human CD8 (+ )T cells to constitutively express Tcf-1 and a TCR specific for the NY-ESO-1 cancer-associated antigen. Co-engineered cells were assessed for their potential for adoptive cellular immunotherapy. METHODS: Tcf-1 mRNA encoding TCF-1B and TCF-1E isoforms, along with GzmB expression were assessed in CD62L (+ )CD57 (−), CD62L (− )CD57 (−), and CD62L (− )CD57 (+ )CD8 (+ )T cells derived from normal donor lymphocytes. The impact of stable Tcf-1B expression on CD8 (+ )T-cell phenotype, anti-tumor activity, and cell-cycle activity was assessed in vitro and in an in vivo tumor xenograft model. RESULTS: TCF-1B and TCF-1E were dynamically regulated during self-renewal, with progeny of recently activated naïve T cells more enriched for TCF-1B mRNA. Constitutive TCF-1B expression improved the survival of TCR-engineered CD8 (+ )T cells upon engagement with tumor cells. Tcf-1B prohibited the acquisition of a GzmB (High) state, and protected T cells from apoptosis associated with elicitation of effector function, and promoted stem cell-like characteristics. CONCLUSIONS: Tcf-1 protects TCR-engineered CD8 (+ )T cells from activation induced cell death by restricting GzmB expression. Our study presents constitutive Tcf-1B expression as a potential means to impart therapeutic T cells with attributes of persistence for durable anti-tumor activity. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s00262-022-03197-2.