Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity
Abstract Background Studies of chimeric antigen receptor (CAR)-T and -Natural killer (NK) cells have shown promising results in treating hematological malignancies. However, there are still obstacles to effectively treating solid tumors. These include the challenges of CAR-T cell homing and infiltra...
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BMC
2025-07-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-025-04461-9 |
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| author | Yuma Fukutani Kenji Kurachi Yu-suke Torisawa Kotoko Miyata Makoto Hayashi Kaoru Sasaki Kodai Saitoh Sono Watanabe Yudai Hasegawa Yoichi Naritomi Yuka Igarashi Kumiko Goto Yuka Sato Noriko Uesugi Hidetaka Murai Tetsuya Sakurai Toru Ozaki Norihiro Tsuneyoshi Masashi Yamada Yuriko Takeno Tomonori Hosoya Fusako Nishigaki Hironobu Kimura Kouichi Tamura |
| author_facet | Yuma Fukutani Kenji Kurachi Yu-suke Torisawa Kotoko Miyata Makoto Hayashi Kaoru Sasaki Kodai Saitoh Sono Watanabe Yudai Hasegawa Yoichi Naritomi Yuka Igarashi Kumiko Goto Yuka Sato Noriko Uesugi Hidetaka Murai Tetsuya Sakurai Toru Ozaki Norihiro Tsuneyoshi Masashi Yamada Yuriko Takeno Tomonori Hosoya Fusako Nishigaki Hironobu Kimura Kouichi Tamura |
| author_sort | Yuma Fukutani |
| collection | DOAJ |
| description | Abstract Background Studies of chimeric antigen receptor (CAR)-T and -Natural killer (NK) cells have shown promising results in treating hematological malignancies. However, there are still obstacles to effectively treating solid tumors. These include the challenges of CAR-T cell homing and infiltration, the presence of immunosuppressive microenvironments, and the potential for antigen escape in solid tumors. To overcome the known limitations of immune cell therapy, we engineered human induced pluripotent stem cell (hiPSC)-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D-DAP10 complex. Methods We introduced the six genes, CCL19, CCR2B, FCGR3A (CD16), IL-15, KLRK1 (NKG2D), and HCST (DAP10), which were controlled under human EF1a promoter, into hiPSCs using the piggyBac system and differentiated them into NK cells. We evaluate the antitumor function, including killing activity, antibody-dependent cytotoxicity, migration ability, and recruitment of dendritic cells. In addition, in vivo antitumor activity was determined by using an orthotopic lung cancer mouse model. Results The gene-engineered hiPSCs expressed all six transgenes, showed normal karyotypes, and were able to differentiate into CD56+ NK cells. The gene-engineered hiPSC-derived NK (eNK) cells showed improvement in viability without additional cytokine supplement in vitro and in vivo. Overexpression of NKG2D complex and high-affinity CD16 enhanced the antitumor function of the eNK cells. Forced expression of CCR2B enhanced eNK cell tumor infiltration. Forced expression of CCL19 endowed the eNK cells with the ability to recruit dendritic cells. We found that the eNK cells were able to lyse HLA-E-expressing tumor cells, but not normal human cells. Moreover, eNK cells demonstrated superior anti-tumor activity in an orthotropic lung cancer mouse model. Conclusion These proof-of-concept studies demonstrate the promise of our eNK cells as a novel adoptive cell therapy product for the treatment of solid tumors. Graphical abstract |
| format | Article |
| id | doaj-art-b62eab7bdc6a43dc94d0cb1b3342e391 |
| institution | Kabale University |
| issn | 1757-6512 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
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| series | Stem Cell Research & Therapy |
| spelling | doaj-art-b62eab7bdc6a43dc94d0cb1b3342e3912025-08-20T03:45:57ZengBMCStem Cell Research & Therapy1757-65122025-07-0116112410.1186/s13287-025-04461-9Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activityYuma Fukutani0Kenji Kurachi1Yu-suke Torisawa2Kotoko Miyata3Makoto Hayashi4Kaoru Sasaki5Kodai Saitoh6Sono Watanabe7Yudai Hasegawa8Yoichi Naritomi9Yuka Igarashi10Kumiko Goto11Yuka Sato12Noriko Uesugi13Hidetaka Murai14Tetsuya Sakurai15Toru Ozaki16Norihiro Tsuneyoshi17Masashi Yamada18Yuriko Takeno19Tomonori Hosoya20Fusako Nishigaki21Hironobu Kimura22Kouichi Tamura23Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Research Division, Kobe Research Institute, HEALIOS K.K.Abstract Background Studies of chimeric antigen receptor (CAR)-T and -Natural killer (NK) cells have shown promising results in treating hematological malignancies. However, there are still obstacles to effectively treating solid tumors. These include the challenges of CAR-T cell homing and infiltration, the presence of immunosuppressive microenvironments, and the potential for antigen escape in solid tumors. To overcome the known limitations of immune cell therapy, we engineered human induced pluripotent stem cell (hiPSC)-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D-DAP10 complex. Methods We introduced the six genes, CCL19, CCR2B, FCGR3A (CD16), IL-15, KLRK1 (NKG2D), and HCST (DAP10), which were controlled under human EF1a promoter, into hiPSCs using the piggyBac system and differentiated them into NK cells. We evaluate the antitumor function, including killing activity, antibody-dependent cytotoxicity, migration ability, and recruitment of dendritic cells. In addition, in vivo antitumor activity was determined by using an orthotopic lung cancer mouse model. Results The gene-engineered hiPSCs expressed all six transgenes, showed normal karyotypes, and were able to differentiate into CD56+ NK cells. The gene-engineered hiPSC-derived NK (eNK) cells showed improvement in viability without additional cytokine supplement in vitro and in vivo. Overexpression of NKG2D complex and high-affinity CD16 enhanced the antitumor function of the eNK cells. Forced expression of CCR2B enhanced eNK cell tumor infiltration. Forced expression of CCL19 endowed the eNK cells with the ability to recruit dendritic cells. We found that the eNK cells were able to lyse HLA-E-expressing tumor cells, but not normal human cells. Moreover, eNK cells demonstrated superior anti-tumor activity in an orthotropic lung cancer mouse model. Conclusion These proof-of-concept studies demonstrate the promise of our eNK cells as a novel adoptive cell therapy product for the treatment of solid tumors. Graphical abstracthttps://doi.org/10.1186/s13287-025-04461-9Human induced pluripotent stem cellGene engineeringNatural killer cellSolid tumorImmunotherapyOff-the-shelf product |
| spellingShingle | Yuma Fukutani Kenji Kurachi Yu-suke Torisawa Kotoko Miyata Makoto Hayashi Kaoru Sasaki Kodai Saitoh Sono Watanabe Yudai Hasegawa Yoichi Naritomi Yuka Igarashi Kumiko Goto Yuka Sato Noriko Uesugi Hidetaka Murai Tetsuya Sakurai Toru Ozaki Norihiro Tsuneyoshi Masashi Yamada Yuriko Takeno Tomonori Hosoya Fusako Nishigaki Hironobu Kimura Kouichi Tamura Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity Stem Cell Research & Therapy Human induced pluripotent stem cell Gene engineering Natural killer cell Solid tumor Immunotherapy Off-the-shelf product |
| title | Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity |
| title_full | Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity |
| title_fullStr | Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity |
| title_full_unstemmed | Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity |
| title_short | Human iPSC-derived NK cells armed with CCL19, CCR2B, high-affinity CD16, IL-15, and NKG2D complex enhance anti-solid tumor activity |
| title_sort | human ipsc derived nk cells armed with ccl19 ccr2b high affinity cd16 il 15 and nkg2d complex enhance anti solid tumor activity |
| topic | Human induced pluripotent stem cell Gene engineering Natural killer cell Solid tumor Immunotherapy Off-the-shelf product |
| url | https://doi.org/10.1186/s13287-025-04461-9 |
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