Journal
CANCER IMMUNOLOGY RESEARCH
Volume 6, Issue 4, Pages 467-480Publisher
AMER ASSOC CANCER RESEARCH
DOI: 10.1158/2326-6066.CIR-17-0207
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Funding
- Swedish Research Council
- Swedish Children's Cancer Society
- Swedish Cancer Society
- Karolinska Institutet
- Swedish Foundation for Strategic Research
- Norwegian Cancer Society
- Norwegian Research Council
- South-Eastern Norway Regional Health Authority
- European Commission Horizon Program [692180-STREAMH2020-TWINN-2015]
- National Science Center, Poland [2014/13/N/NZ6/02081]
- Stiftelsen Kristian Gerhard Jebsen
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Natural killer (NK) cells hold potential as a source of allogeneic cytotoxic effector cells for chimeric antigen receptor (CAR)-mediated therapies. Here, we explored the feasibility of transfecting CAR-encoding mRNA into primary NK cells and investigated how the intrinsic potential of discrete NK-cell subsets affects retargeting efficiency. After screening five second- and third-generation anti-CD19 CAR constructs with different signaling domains and spacer regions, a third-generation CAR with the CH2-domain removed was selected based on its expression and functional profiles. Kinetics experiments revealed that CAR expression was optimal after 3 days of IL15 stimulation prior to transfection, consistently achieving over 80% expression. CAR-engineered NK cells acquired increased degranulation toward CD19(+) targets, and maintained their intrinsic degranulation response toward CD19(-) K562 cells. The response of redirected NK-cell subsets against CD19(+) targets was dependent on their intrinsic thresholds for activation determined through both differentiation and education by killer cell immunoglobulin-like receptors (KIR) and/or CD94/NKG2A binding to self HLA class I and HLA-E, respectively. Redirected primary NK cells were insensitive to inhibition through NKG2A/HLA-E interactions but remained sensitive to inhibition through KIR depending on the amount of HLA class I expressed on target cells. Adaptive NK cells, expressing NKG2C, CD57, and self-HLA-specific KIR(s), displayed superior ability to kill CD19(+), HLA low, or mismatched tumor cells. These findings support the feasibility of primary allogeneic NK cells for CAR engineering and highlight a need to consider NK-cell diversity when optimizing efficacy of cancer immunotherapies based on CAR-expressing NK cells. (C) 2018 AACR.
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