{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T15:10:38Z","timestamp":1788793838139,"version":"build-2803163510"},"reference-count":35,"publisher":"Oxford University Press (OUP)","issue":"6","license":[{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"name":"ChadTough DIPG Foundation and the Purdue Institute for Cancer Research"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,6,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Reproducible and efficient genetic engineering of human NK cells remains a primary challenge to next-generation chimeric antigen receptor (CAR) therapies, specifically for larger constructs with multiple plasmids and transgenes. Lentiviral vectors (LVs) pseudotyped with baboon envelope (BaEV) outperform vesicular stomatitis virus G protein in transducing hematopoietic stem cells, T cells, B cells, and NK cells, but suffer from fusogenic toxicity in producer cells and lower virion yields (BaEV-TR). Here, we engineer an integrated viral receptor platform that overcomes these constraints and enables transduction of freshly isolated, resting NK cells. First, we generated a Sleeping Beauty\u2013engineered HEK293T packaging cell line stably expressing the nonfusogenic BaEV-TR glycoprotein, enabling consistent and scalable production of high titer BaEV-TR\u2013pseudotyped LVs. Using this system, we generated LVs encoding SLC1A5 and a large (\u223c9.6\u2009kb) multifunctional CD73-GD2-NKG2D CAR (mCAR). Second, we augmented SLC1A5 expression in freshly-isolated human NK cells using PDK1 inhibitor BX-795 and BaEV-TR-pseudotyped LVs. Rapid SLC1A5 overexpression in NK cells enabled transduction of resting cells without prior expansion and significantly increased NK cell susceptibility to mCAR delivery, achieving robust expression of the complex CAR cassette and preserving NK cell phenotype, cytokine production, degranulation, proliferation, and cytotoxicity against aggressive solid tumor cell targets. This combined approach directly addresses major bottlenecks in CAR-NK cell manufacturing, providing a modular platform for programming NK cells with challenging and large payloads.<\/jats:p>","DOI":"10.1093\/jimmun\/vkag133","type":"journal-article","created":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T04:59:57Z","timestamp":1782449997000},"source":"Crossref","is-referenced-by-count":0,"title":["SLC1A5 augmentation bypasses NK cell transduction barriers to deliver complex CAR payloads"],"prefix":"10.1093","volume":"215","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-5974-7649","authenticated-orcid":false,"given":"Soumyajit","family":"Das","sequence":"first","affiliation":[{"name":"Department of Industrial and Molecular Pharmaceutics, Purdue University , West Lafayette, IN,","place":["United States"]},{"name":"Institute for Cancer Research, Purdue University , West Lafayette, IN,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khanh Ha","family":"Nguyen","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Purdue University , West Lafayette, IN,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tyler","family":"Fritsch","sequence":"additional","affiliation":[{"name":"Department of Industrial and Molecular Pharmaceutics, Purdue University , West Lafayette, IN,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shambhavi","family":"Borde","sequence":"additional","affiliation":[{"name":"Department of Industrial and Molecular Pharmaceutics, Purdue University , West Lafayette, IN,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Els","family":"Verhoeyen","sequence":"additional","affiliation":[{"name":"INSERM U1111, Universit\u00e9 de Lyon , Lyon,","place":["France"]},{"name":"INSERM U1065 , Lyon,","place":["France"]},{"name":"Universit\u00e9 C\u00f4te d\u2019Azur, INSERM, C3M , Nice,","place":["France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5118-2455","authenticated-orcid":false,"given":"Sandro","family":"Matosevic","sequence":"additional","affiliation":[{"name":"Department of Industrial and Molecular Pharmaceutics, Purdue University , West Lafayette, IN,","place":["United States"]},{"name":"Institute for Cancer Research, Purdue University , West Lafayette, IN,","place":["United States"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"key":"2026090710425929700_vkag133-B1","doi-asserted-by":"crossref","first-page":"286","DOI":"10.3389\/fimmu.2015.00286","article-title":"Present and future of allogeneic natural killer cell therapy","volume":"6","author":"Lim","year":"2015","journal-title":"Front 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Mater"},{"key":"2026090710425929700_vkag133-B5","doi-asserted-by":"crossref","first-page":"2873","DOI":"10.3389\/fimmu.2019.02873","article-title":"Efficient and robust NK-cell transduction with baboon envelope pseudotyped lentivector","volume":"10","author":"Colamartino","year":"2019","journal-title":"Front Immunol"},{"key":"2026090710425929700_vkag133-B6","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1016\/j.omtm.2021.01.008","article-title":"Systematic improvements in lentiviral transduction of primary human natural killer cells undergoing ex vivo expansion","volume":"20","author":"Allan","year":"2021","journal-title":"Mol Ther Methods Clin Dev"},{"key":"2026090710425929700_vkag133-B7","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1038\/s41434-024-00454-0","article-title":"Development of KoRV-pseudotyped lentiviral vectors for efficient gene transfer into freshly isolated immune cells","volume":"31","author":"Renner","year":"2024","journal-title":"Gene 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HSCs","volume":"124","author":"Girard-Gagnepain","year":"2014","journal-title":"Blood"},{"key":"2026090710425929700_vkag133-B11","doi-asserted-by":"publisher","first-page":"12365","DOI":"10.1038\/s41598-023-39597-2","article-title":"Optimized conditions for gene transduction into primary immune cells using viral vectors","volume":"13","author":"Kim","year":"2023","journal-title":"Sci Rep"},{"key":"2026090710425929700_vkag133-B12","doi-asserted-by":"crossref","first-page":"1548630","DOI":"10.3389\/fimmu.2025.1548630","article-title":"Transduction of \u03b3\u03b4 T cells with baboon envelope pseudotyped lentiviral vector encoding chimeric antigen receptors for translational and clinical applications","volume":"16","author":"Pinot","year":"2025","journal-title":"Front Immunol"},{"key":"2026090710425929700_vkag133-B13","doi-asserted-by":"crossref","first-page":"2478","DOI":"10.1111\/jth.13520","article-title":"Baboon envelope pseudotyped lentiviral vectors efficiently transduce human B 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