An improved approach to generate IL-15+/+/TGFßR2-/- iPSC-derived natural killer cells using TALEN.
Cell Rep Methods
; 4(9): 100857, 2024 Sep 16.
Article
in En
| MEDLINE
| ID: mdl-39260365
ABSTRACT
We present a TALEN-based workflow to generate and maintain dual-edited (IL-15+/+/TGFßR2-/-) 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 TGFßR2 in immune cells can enhance resistance to the suppressive TGF-ß signaling in the tumor microenvironment, we develop a customized medium containing Nodal that can maintain the pluripotency of iPSCs with TGFßR2 KO, enabling banking of these iPSC clones. Ultimately, we show that the dual-edited IL-15+/+/TGFßR2-/- iPSCs can be efficiently differentiated into NK cells that show enhanced autonomous growth and are resistant to the suppressive TGF-ß signaling.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Killer Cells, Natural
/
Interleukin-15
/
Induced Pluripotent Stem Cells
/
Receptor, Transforming Growth Factor-beta Type II
Limits:
Humans
Language:
En
Journal:
Cell Rep Methods
Year:
2024
Document type:
Article
Country of publication: