Session: 703. Adoptive Immunotherapy: Mechanisms and New Approaches: Poster III
Hematology Disease Topics & Pathways:
Biological, Diseases, iPSCs, Therapies, Biological Processes, Technology and Procedures, immune cells, immunotherapy, Cell Lineage, gene editing, NK cells, flow cytometry, stem cells, NGS, microenvironment
In this study, we used CRISPR/Cas12a to generate edited iPSC lines that were differentiated into CD56+ iNK cells. Specifically, we generated TGFβR2-/-, CISH-/-, and TGFβR2-/-/CISH-/- iPSC clones with bi-allelic gene disruption confirmed by next generation sequencing. Importantly, we also confirmed that the edited clones were pluripotent. In particular, a minimum of 3 clones from each genotype were differentiated to CD56+ iNK cells. After differentiation, >90% of the cells expressed CD56 for all genotypes. Additionally, we observed the expression of canonical natural killer cell markers such as CD16, NKG2A, KIRs, NKp46, NKp44, and NKp30 within this CD56+ population.
We tested the effector function of TGFβR2-/-, CISH-/-, and TGFβR2-/-/CISH-/- iNKs in a variety of molecular and functional assays, including a spheroid killing assay and an in vitro serial killing assay. For example, we utilized a SK-OV-3 spheroid killing assay to determine the intrinsic ability for the iNK cells to kill tumor targets following the differentiation process. TGFβR2-/-, CISH-/-, and TGFβR2-/-/CISH-/- iNKs reduce the size of SK-OV-3 ovarian tumor spheroids more effectively than control iNK cells in the presence of exogenous TGF-β.
In conclusion, we have established an iPSC editing platform that can generate a near infinite supply of natural killer cells with enhanced tumor killing function, paving the way for future off-the-shelf cell therapies for application to broad oncology indications.
Disclosures: Moon: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Chin: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Burden: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Sexton: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Wasko: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Nasser: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Antony: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Wong: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Borges: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Morgan: Editas Medicine: Current Employment, Current equity holder in publicly-traded company. Welstead: Editas Medicine: Current Employment, Current equity holder in publicly-traded company.
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