1. Engineering Stage-Specific Developmental Cues to Generate iPSC-derived, Non-allogenic, Chimeric Antigen Receptor T cells for Immunotherapy
- Author
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Yoo, Sang Pil, Crooks, Gay M1, Yoo, Sang Pil, Yoo, Sang Pil, Crooks, Gay M1, and Yoo, Sang Pil
- Abstract
Chimeric antigen receptor (CAR) T cell therapy has demonstrated remarkable results in otherwise treatment-refractory diseases, most notably for B cell malignancies. However, there are several challenges to autologous CAR T therapy that limit its accessibility and widespread adoption. First, the de novo generation of each treatment can lead to CAR T cells that exhibit varying clinical responses, in part due to inherent variations in the T cell composition of each product. The variable outcomes are compounded by complex manufacturing processes and T cell expansion procedures that can produce a prematurely exhausted and less efficacious CAR T product. These challenges contribute to cancer relapse rates as high as 57-66% after treatment. Thus, there is a growing interest in generating functionally enhanced CAR T cells in vitro from a self-renewing source of induced pluripotent stem cells (iPSCs). iPSC-engineered T cell therapy would ensure immediate access to treatment that delivers a more durable and consistent clinical response.Unlike peripheral blood T cells, the introduction or the perturbation of biologically active genes can pose serious challenges to the in vitro differentiation of iPSCs. Inappropriate signaling or developmental cues at critical stages of T cell development can result in a block in differentiation or diversion to an undesired lineage. We and others have identified two critical biologic barriers to the generation of iPSC-derived CAR T cells: (1) expression of the CAR in early, T cell precursors diverts differentiation towards the innate lineage, instead of the conventional T cell pathway, and (2) Removal of endogenous T cell receptor (TCR) to address the risk of alloreactivity halts T cell development due to the absence of positive selection. This dissertation introduces a novel CRISPR/Cas9 editing strategy that aim to address both CAR-mediated innate diversion and rescue of positive selection in the absence of endogenous TCR. We report the genera
- Published
- 2024