Efficient, fratricide free non-viral engineering of CD70-targeted CAR NK cells for hematologic and solid tumor immunotherapy

Jae Woong Chang; Joshua Krueger; Timothy D. Folsom; Joseph G. Skeate; Erin M. Stelljes; Walker S. Lahr; Nicholas J. Slipek; Emily J. Pomeroy; Beau R. Webber; Branden S. Moriarity

doi:10.1016/j.omta.2026.201734

Efficient, fratricide free non-viral engineering of CD70-targeted CAR NK cells for hematologic and solid tumor immunotherapy

Pediatric Hematology/Oncology

Research output: Contribution to journal › Article › peer-review

Abstract

Natural killer (NK) cells are a component of the innate immune system with potential safety advantages over T cells for adoptive cell therapy (ACT). However, NK cells lack the potency and persistence needed for tumor clearance. Here, we report an efficient and cost-effective non-viral method for engineering CD70 targeted CAR NK cells using a hyperactive TcBuster DNA transposon system while simultaneously knocking out CD70 via CRISPR-Cas9 to prevent fratricide. CD70 CAR NK cells engineered in this manner display improved efficacy in killing acute myeloid lymphoma (AML) cells and renal cell carcinoma (RCC) cells in vitro and in vivo. Incorporating soluble IL-15 cytokine armoring into the transposon cassette further improves survival and expansion in vivo. Our findings demonstrate a novel application of non-viral transposon-based cell engineering combined with CRISPR-Cas9 knockout to produce a clinically scalable product for ACT against multiple cancer types.

Original language	English (US)
Article number	201734
Journal	Molecular Therapy Advances
Volume	34
Issue number	2
DOIs	https://doi.org/10.1016/j.omta.2026.201734
State	Published - Jun 11 2026

Bibliographical note

Publisher Copyright:
© 2026 University of Minnesota. Published by Elsevier Inc. on behalf of The American Society of Gene and Cell Therapy. This is an open access article under the CC BY-NC-ND license. http://creativecommons.org/licenses/by-nc-nd/4.0/

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

CRISPR-Cas9 CD70
TcBuster
acute myeloid leukemia
chimeric antigen receptor
fratricide
natural killer cells
renal cell carcinoma

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10.1016/j.omta.2026.201734

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Chang, J. W., Krueger, J., Folsom, T. D., Skeate, J. G., Stelljes, E. M., Lahr, W. S., Slipek, N. J., Pomeroy, E. J., Webber, B. R., & Moriarity, B. S. (2026). Efficient, fratricide free non-viral engineering of CD70-targeted CAR NK cells for hematologic and solid tumor immunotherapy. Molecular Therapy Advances, 34(2), Article 201734. https://doi.org/10.1016/j.omta.2026.201734

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abstract = "Natural killer (NK) cells are a component of the innate immune system with potential safety advantages over T cells for adoptive cell therapy (ACT). However, NK cells lack the potency and persistence needed for tumor clearance. Here, we report an efficient and cost-effective non-viral method for engineering CD70 targeted CAR NK cells using a hyperactive TcBuster DNA transposon system while simultaneously knocking out CD70 via CRISPR-Cas9 to prevent fratricide. CD70 CAR NK cells engineered in this manner display improved efficacy in killing acute myeloid lymphoma (AML) cells and renal cell carcinoma (RCC) cells in vitro and in vivo. Incorporating soluble IL-15 cytokine armoring into the transposon cassette further improves survival and expansion in vivo. Our findings demonstrate a novel application of non-viral transposon-based cell engineering combined with CRISPR-Cas9 knockout to produce a clinically scalable product for ACT against multiple cancer types.",

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AU - Chang, Jae Woong

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AU - Folsom, Timothy D.

AU - Skeate, Joseph G.

AU - Stelljes, Erin M.

AU - Lahr, Walker S.

AU - Slipek, Nicholas J.

AU - Pomeroy, Emily J.

AU - Webber, Beau R.

AU - Moriarity, Branden S.

N1 - Publisher Copyright: © 2026 University of Minnesota. Published by Elsevier Inc. on behalf of The American Society of Gene and Cell Therapy. This is an open access article under the CC BY-NC-ND license. http://creativecommons.org/licenses/by-nc-nd/4.0/

PY - 2026/6/11

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N2 - Natural killer (NK) cells are a component of the innate immune system with potential safety advantages over T cells for adoptive cell therapy (ACT). However, NK cells lack the potency and persistence needed for tumor clearance. Here, we report an efficient and cost-effective non-viral method for engineering CD70 targeted CAR NK cells using a hyperactive TcBuster DNA transposon system while simultaneously knocking out CD70 via CRISPR-Cas9 to prevent fratricide. CD70 CAR NK cells engineered in this manner display improved efficacy in killing acute myeloid lymphoma (AML) cells and renal cell carcinoma (RCC) cells in vitro and in vivo. Incorporating soluble IL-15 cytokine armoring into the transposon cassette further improves survival and expansion in vivo. Our findings demonstrate a novel application of non-viral transposon-based cell engineering combined with CRISPR-Cas9 knockout to produce a clinically scalable product for ACT against multiple cancer types.

AB - Natural killer (NK) cells are a component of the innate immune system with potential safety advantages over T cells for adoptive cell therapy (ACT). However, NK cells lack the potency and persistence needed for tumor clearance. Here, we report an efficient and cost-effective non-viral method for engineering CD70 targeted CAR NK cells using a hyperactive TcBuster DNA transposon system while simultaneously knocking out CD70 via CRISPR-Cas9 to prevent fratricide. CD70 CAR NK cells engineered in this manner display improved efficacy in killing acute myeloid lymphoma (AML) cells and renal cell carcinoma (RCC) cells in vitro and in vivo. Incorporating soluble IL-15 cytokine armoring into the transposon cassette further improves survival and expansion in vivo. Our findings demonstrate a novel application of non-viral transposon-based cell engineering combined with CRISPR-Cas9 knockout to produce a clinically scalable product for ACT against multiple cancer types.

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KW - acute myeloid leukemia

KW - chimeric antigen receptor

KW - fratricide

KW - natural killer cells

KW - renal cell carcinoma

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Efficient, fratricide free non-viral engineering of CD70-targeted CAR NK cells for hematologic and solid tumor immunotherapy

Abstract

Bibliographical note

UN SDGs

Keywords

Publisher link

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