- Wang, Zhiqiang;
- McWilliams-Koeppen, Helen P;
- Reza, Hernan;
- Ostberg, Julie R;
- Chen, Wuyang;
- Wang, Xiuli;
- Huynh, Christian;
- Vyas, Vibhuti;
- Chang, Wen-Chung;
- Starr, Renate;
- Wagner, Jamie R;
- Aguilar, Brenda;
- Yang, Xin;
- Wu, Xiwei;
- Wang, Jinhui;
- Chen, Wei;
- Koelker-Wolfe, Ellery;
- Seet, Christopher S;
- Montel-Hagen, Amélie;
- Crooks, Gay M;
- Forman, Stephen J;
- Brown, Christine E
Unlimited generation of chimeric antigen receptor (CAR) T cells from human-induced pluripotent stem cells (iPSCs) is an attractive approach for "off-the-shelf" CAR T cell immunotherapy. Approaches to efficiently differentiate iPSCs into canonical αβ T cell lineages, while maintaining CAR expression and functionality, however, have been challenging. We report that iPSCs reprogramed from CD62L+ naive and memory T cells followed by CD19-CAR engineering and 3D-organoid system differentiation confers products with conventional CD8αβ-positive CAR T cell characteristics. Expanded iPSC CD19-CAR T cells showed comparable antigen-specific activation, degranulation, cytotoxicity, and cytokine secretion compared with conventional CD19-CAR T cells and maintained homogeneous expression of the TCR derived from the initial clone. iPSC CD19-CAR T cells also mediated potent antitumor activity in vivo, prolonging survival of mice with CD19+ human tumor xenografts. Our study establishes feasible methodologies to generate highly functional CAR T cells from iPSCs to support the development of "off-the-shelf" manufacturing strategies.