3D-organoid culture supports differentiation of human CAR+ iPSCs into highly functional CAR T cells

• DOI: 10.1016/j.stem.2022.02.009
• Publication Date: 2022-03-11T15:32:03Z
• Authors: Zhiqiang Wang; Helen P. McWillia...; Hernan Reza; Julie R. Ostberg; Wuyang Chen; Xiuli Wang; Christian Huynh; Vibhuti Vyas; Wen-Chung Chang; Renate Starr; Jamie R. Wagner; Brenda Aguilar; Xin Yang; Xiwei Wu; Jinhui Wang; Wei Chen; Ellery Koelker-Wolfe; Christopher S. Seet; Amélie Montel-Hagen; Gay M. Crooks; Stephen J. Forman; Christine E. Brown
• Keywords: clone (Java method)

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.