Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells
Natural killer (NK) cells are pivotal in immunosurveillance and hold great potential for immunotherapy due to their ability to target malignant cells. Their low risk of causing graft-versus-host disease (GvHD) post-allogenic transplantation underscores their potential as an off-the shelf cellular th...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-06-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050125000683 |
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| author | Taha B. Hayal Aman A. Mulla David S.J. Allan Brynn B. Duncan Saanika Joshi So Gun Hong Rafet Basar Katayoun Rezvani Richard W. Childs Chuanfeng Wu Cynthia E. Dunbar |
| author_facet | Taha B. Hayal Aman A. Mulla David S.J. Allan Brynn B. Duncan Saanika Joshi So Gun Hong Rafet Basar Katayoun Rezvani Richard W. Childs Chuanfeng Wu Cynthia E. Dunbar |
| author_sort | Taha B. Hayal |
| collection | DOAJ |
| description | Natural killer (NK) cells are pivotal in immunosurveillance and hold great potential for immunotherapy due to their ability to target malignant cells. Their low risk of causing graft-versus-host disease (GvHD) post-allogenic transplantation underscores their potential as an off-the shelf cellular therapy tool. Advances in genetic engineering focus on improving NK targeting, persistence, and fitness. However, NK cells pose challenges for lentiviral transduction, which are clinically relevant and safe. In this study, we identified Poloxamer 407 (P407) as a novel transduction enhancer for rhesus macaque (RM) and human NK cells. We found that P407 significantly improved transduction efficiency, achieving up to 60% in expanded RM NK cells, without compromising cell viability or functionality. Additionally, P407 facilitated the expression of anti-CD20 chimeric antigen receptors (CARs) with or without interleukin (IL)-15. In a xenograft mouse model, CAR-IL15 NK cells demonstrated superior anti-tumor activity, and maintained higher clonal diversity tracked by genetic barcoding compared to CAR-NK cells lacking IL-15 in vivo. Additionally, in human NK cells, P407 combined with the TBK1/IKKε inhibitor, BX795, further improved lentivirus-mediated transduction. This study is the first to engineer NK cells from a clinically relevant rhesus macaque model in an adaptive cell therapy context and highlights P407’s potential as a transduction enhancer. |
| format | Article |
| id | doaj-art-8efa3540a1a04e0189e1ccd2aa2169c0 |
| institution | OA Journals |
| issn | 2329-0501 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj-art-8efa3540a1a04e0189e1ccd2aa2169c02025-08-20T02:27:15ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-06-0133210147310.1016/j.omtm.2025.101473Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cellsTaha B. Hayal0Aman A. Mulla1David S.J. Allan2Brynn B. Duncan3Saanika Joshi4So Gun Hong5Rafet Basar6Katayoun Rezvani7Richard W. Childs8Chuanfeng Wu9Cynthia E. Dunbar10Translational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USATranslational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USALaboratory of Transplantation Immunotherapy, Cellular and Molecular Therapeutics Branch, NHLBI, NIH, Bethesda, MD 20892, USATranslational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USATranslational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USATranslational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USADepartment of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USADepartment of Stem Cell Transplantation and Cellular Therapy, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USALaboratory of Transplantation Immunotherapy, Cellular and Molecular Therapeutics Branch, NHLBI, NIH, Bethesda, MD 20892, USATranslational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USA; Corresponding author: Chuanfeng Wu, Translational Stem Cell Biology Branch, National Heart, Lung, and Blood Institute, NIH Building 10 CRC, Room 5E-3288, 10 Center Drive, Bethesda, MD 20892, USA.Translational Stem Cell Biology Branch, NHLBI, NIH, Bethesda, MD 20892, USA; Corresponding author: Cynthia E. Dunbar, Translational Stem Cell Biology Branch, National Heart, Lung, and Blood Institute, NIH Building 10 CRC, Room 5E-3332, 10 Center Drive, Bethesda, MD 20892, USA.Natural killer (NK) cells are pivotal in immunosurveillance and hold great potential for immunotherapy due to their ability to target malignant cells. Their low risk of causing graft-versus-host disease (GvHD) post-allogenic transplantation underscores their potential as an off-the shelf cellular therapy tool. Advances in genetic engineering focus on improving NK targeting, persistence, and fitness. However, NK cells pose challenges for lentiviral transduction, which are clinically relevant and safe. In this study, we identified Poloxamer 407 (P407) as a novel transduction enhancer for rhesus macaque (RM) and human NK cells. We found that P407 significantly improved transduction efficiency, achieving up to 60% in expanded RM NK cells, without compromising cell viability or functionality. Additionally, P407 facilitated the expression of anti-CD20 chimeric antigen receptors (CARs) with or without interleukin (IL)-15. In a xenograft mouse model, CAR-IL15 NK cells demonstrated superior anti-tumor activity, and maintained higher clonal diversity tracked by genetic barcoding compared to CAR-NK cells lacking IL-15 in vivo. Additionally, in human NK cells, P407 combined with the TBK1/IKKε inhibitor, BX795, further improved lentivirus-mediated transduction. This study is the first to engineer NK cells from a clinically relevant rhesus macaque model in an adaptive cell therapy context and highlights P407’s potential as a transduction enhancer.http://www.sciencedirect.com/science/article/pii/S2329050125000683natural killer cell, NK cellPoloxamer 407lentiviral transductionCAR-NKIL-15barcode clonal tracking |
| spellingShingle | Taha B. Hayal Aman A. Mulla David S.J. Allan Brynn B. Duncan Saanika Joshi So Gun Hong Rafet Basar Katayoun Rezvani Richard W. Childs Chuanfeng Wu Cynthia E. Dunbar Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells Molecular Therapy: Methods & Clinical Development natural killer cell, NK cell Poloxamer 407 lentiviral transduction CAR-NK IL-15 barcode clonal tracking |
| title | Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells |
| title_full | Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells |
| title_fullStr | Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells |
| title_full_unstemmed | Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells |
| title_short | Optimization of lentiviral delivery of barcoded anti-CD20 chimeric antigen receptors into rhesus macaque and human natural killer cells |
| title_sort | optimization of lentiviral delivery of barcoded anti cd20 chimeric antigen receptors into rhesus macaque and human natural killer cells |
| topic | natural killer cell, NK cell Poloxamer 407 lentiviral transduction CAR-NK IL-15 barcode clonal tracking |
| url | http://www.sciencedirect.com/science/article/pii/S2329050125000683 |
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