1. An improved approach to generate IL-15+/+/TGFβR2-/- iPSC-derived natural killer cells using TALEN.
- Author
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Chen, An-Ping, Gao, Peng, Lin, Liang, Ashok, Preeti, He, Hongzhi, Ma, Chao, Zou, David, Allain, Vincent, Boyne, Alex, Juillerat, Alexandre, Duchateau, Philippe, Rath, Armin, Teper, Daniel, Arulanandam, Antonio, Chang, Hao-Ming, Eyquem, Justin, and Li, Wei
- Subjects
B2M ,CP: cancer biology ,GUIDE-seq ,TALEN ,TGFβR2 ,anti-tumor ,beta-2-microglobulin ,gene editing ,human induced pluripotent stem cells ,iPSC-derived cell therapies ,iPSCs ,interleukin-15 ,natural killer cells ,Induced Pluripotent Stem Cells ,Killer Cells ,Natural ,Interleukin-15 ,Humans ,Receptor ,Transforming Growth Factor-beta Type II ,Cell Differentiation ,Transcription Activator-Like Effector Nucleases ,Gene Editing - Abstract
We present a TALEN-based workflow to generate and maintain dual-edited (IL-15+/+/TGFβR2-/-) iPSCs that produce enhanced iPSC-derived natural killer (iNK) cells for cancer immunotherapy. It involves using a cell lineage promoter for knocking in (KI) gene(s) to minimize the potential effects of expression of any exogenous genes on iPSCs. As a proof-of-principle, we KI IL-15 under the endogenous B2M promoter and show that it results in high expression of the sIL-15 in iNK cells but minimal expression in iPSCs. Furthermore, given that it is known that knockout (KO) of TGFβR2 in immune cells can enhance resistance to the suppressive TGF-β signaling in the tumor microenvironment, we develop a customized medium containing Nodal that can maintain the pluripotency of iPSCs with TGFβR2 KO, enabling banking of these iPSC clones. Ultimately, we show that the dual-edited IL-15+/+/TGFβR2-/- iPSCs can be efficiently differentiated into NK cells that show enhanced autonomous growth and are resistant to the suppressive TGF-β signaling.
- Published
- 2024