Chimeric synthetic T cell receptor and antigen receptor receptors enable high-sensitivity T cell receptor-negative CD8αβ-CD3-positive T-cell development from human-induced pluripotent stem cells.

Abstract

Induced pluripotent stem cell (iPSC)-derived T cells hold great promise for scalable and standardized allogeneic immunotherapies but face critical challenges in differentiation, antigen sensitivity and inaccessibility to intracellular tumor-associated antigens, particularly when engineered with chimeric antigen receptors (CARs). Here, we introduce iPSC with synthetic T cell receptor (TCR) and antigen receptor (STAR) structure, which fuses a single-chain variable fragment (scFv) to the constant regions of TCRαβ. Using a Wilms tumor 1 (WT1)-specific STAR construct, we demonstrate that STAR-iPSC-derived T cells (STAR-iT cells) restore CD3 expression to levels similar to TCR, thereby supporting efficient CD8αβ⁺ CD3⁺ iPSC-T cell development in the absence of TCR. STAR-iT cells exhibit superior antigen sensitivity compared to CAR-based counterparts. STAR-iT cells display robust cytotoxicity against WT1-expressing tumor cells, enhanced cytokine production and efficient in vitro expansion. Our findings establish STAR as a scFv-based, next-generation chimeric receptor format and advance the development of universal, off-the-shelf iT cell immunotherapies.