T Lymphopoiesis from Pluripotent Stem Cells by Defined Transcription Factors at Single Cell Resolution

To date, preferentially regenerating T lymphopoiesis in vivo from pluripotent stem cells (PSC) remains a practical challenge. Here we documented that synergistic expression of Runx1 and Hoxa9 during endothelial to hematopoietic transition stage preferentially generated hematopoietic progenitors capable of homing to thymus and outputting full T lymphopoiesis in primary and secondary recipients. Single-cell transcriptome and functional analyses illustrated the cellular trajectory of T lineage induction from PSC, unveiling the T-lineage specification determined at as early as hemogenic endothelial cell stage and the bona fide pre-thymic progenitors with thymus-homing traits. The mature iT cells distributed normally in central and peripheral lymphoid organs and exhibited abundant TCRαβ repertoire. Furthermore, the TCR-edited PSC produced functional tumor-specific-TCR-T cells in vivo that effectively eradicated tumor cells and transformed to memory cells. This study provides insight into preferential and complete lymphopoiesis from PSC.