iPSC-derived Trimodal T cells engineered with CAR, TCR, and hnCD16 modalities can overcome antigen escape in heterogenous tumors

Although chimeric antigen receptor (CAR) T cells have demonstrated remarkable therapeutic activity in hematopoietic malignancies, antigen escape and tumor heterogeneity have in part impeded the durable efficacy of CAR T cells and their extension into successful solid tumor treatment. To address these challenges, induced pluripotent stem cell (iPSC) - derived T (iT) cells were specifically engineered to uniformly express both CAR and T cell receptors (TCR), enabling targeting of both surface and intracellular antigens, respectively, along with a high-affinity, non-cleavable variant of CD16a (hnCD16) to support antibody-dependent cell cytotoxicity (ADCC) when combined with therapeutic antibodies. Co-expression of each of these unique anti-tumor targeting strategies on engineered iT cells enabled independent and antigen-specific tumor cell killing across a diverse set of liquid and solid tumors. In heterogenous tumor models, coactivation of two or more of these effector modalities was required for measurable anti-tumor efficacy, with all three displaying maximal efficacy. These data highlight the therapeutic potential of an off-the-shelf engineered iPSC-derived Trimodal T cell expressing CAR, TCR, and hnCD16 to combat difficult to treat heterogeneous tumors. RNA were isolated from cryo vials of human Neutrophils, CD56+ NK cells, Monocytes, Macrophages, CD8+ T cells (commercially available) and human TiPSC, iTs with 3 different engineering designs (generated in house as described in M&M).