Abstract 3613: Screening iPSC lines for optimal characteristics of differentiation into immune effector cells for clinical programs

Abstract

Abstract Engineered Induced Pluripotent Stem Cells (iPSCs) are the core foundational technology for Century Therapeutics. In our constant effort to identify iPSC lines with diverse functional activity, we have derived a large panel of clinical-grade peripheral blood mononuclear cell (PBMC)-derived and gamma-delta T cell-derived IPSC lines (PiPSCs and TiPSCs, respectively) from multiple donors. These lines have been screened by multiple criteria to select the top lines for clinical development in our iPSC-derived NK (iNK) and T cell (iT) programs. Initial screening included in-depth genomic and transcriptomic analysis of PiPSC and TiPSC lines to eliminate those with any unwanted genetic abnormalities, as well as to build multi-omics datasets for correlating gene polymorphisms and transcript variation to function. Next, all lines were differentiated to iNK or iT effector cells, and phenotypically characterized throughout the differentiation process. In those studies, 86.5% of PiPSCs and 69.2% of TiPSCs lines were compatible with our protocols and able to be successfully differentiated and acquire NK cell and gamma delta T cell phenotypes respectively. Yield and lineage commitment (%CD56+ for NK cells and %CD3+ for T cells) variability was found to be higher across clones than donors in this initial analysis. Finally, we evaluated in vitro cytotoxicity, cytokine secretion, and persistence for all clones that met differentiation thresholds (>90% lineage commitment). As part of that evaluation, 39 PiPSCs lines were differentiated to iNK cells. Cytotoxicity against tumor lines was observed to be consistently between 85-100% at an effector to target ratio of 1:1. Although no trends in functionality were found to be associated with the PBMC donor, iPSC lines derived from CD34+ enriched PBMCs exhibited higher levels of innate killing as compared to those derived from bulk PBMCs. For this presentation, we will present genetic, transcriptomic, and phenotypic correlations with favorable differentiation and functional efficacy and illustrate how our screening process enables efficient selection of founder lines with the highest therapeutic potential for allogeneic cell therapies. Citation Format: Barry A. Morse, Amarin Cogburn, Alex Chialastri, Matthew S. Hall, Ohad Manor, Andriana Lebid, Dae Hwan Kim, Luis Cocka, Daniel J. Perry, Ciara Budd, Aarti Kolluri, Liam Campion, Toshinobu Nishimura, Michael Naso, Hyam Levitsky. Screening iPSC lines for optimal characteristics of differentiation into immune effector cells for clinical programs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3613.