{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,25]],"date-time":"2026-10-25T11:57:55Z","timestamp":1792929475760,"version":"4.5.0"},"reference-count":33,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2023,8,29]],"date-time":"2023-08-29T00:00:00Z","timestamp":1693267200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Promab Biotechnologies"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Natural killer (NK) cells are cytotoxic lymphocytes that are critical for the innate immune system. Engineering NK cells with chimeric antigen receptors (CARs) allows CAR-NK cells to target tumor antigens more effectively. In this report, we present novel CAR mRNA-LNP (lipid nanoparticle) technology to effectively transfect NK cells expanded from primary PBMCs and to generate functional CAR-NK cells. CD19-CAR mRNA and BCMA-CAR mRNA were embedded into LNPs that resulted in 78% and 95% CAR expression in NK cells, respectively. BCMA-CAR-NK cells after transfection with CAR mRNA-LNPs killed multiple myeloma RPMI8226 and MM1S cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner in vitro. In addition, CD19-CAR-NK cells generated with CAR mRNA-LNPs killed Daudi and Nalm-6 cells and secreted IFN-gamma and Granzyme B in a dose-dependent manner. Both BCMA-CAR-NK and CD19-CAR-NK cells showed significantly higher cytotoxicity, IFN-gamma, and Granzyme B secretion compared with normal NK cells. Moreover, CD19-CAR-NK cells significantly blocked Nalm-6 tumor growth in vivo. Thus, non-viral delivery of CAR mRNA-LNPs can be used to generate functional CAR-NK cells with high anti-tumor activity.<\/jats:p>","DOI":"10.3390\/ijms241713364","type":"journal-article","created":{"date-parts":[[2023,8,29]],"date-time":"2023-08-29T08:26:52Z","timestamp":1693297612000},"page":"13364","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":67,"title":["CAR-NK Cells Generated with mRNA-LNPs Kill Tumor Target Cells In Vitro and In Vivo"],"prefix":"10.3390","volume":"24","author":[{"given":"Vita","family":"Golubovskaya","sequence":"first","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Sienkiewicz","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinying","family":"Sun","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-6874-6541","authenticated-orcid":false,"given":"Shiming","family":"Zhang","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanwei","family":"Huang","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hua","family":"Zhou","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hizkia","family":"Harto","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shirley","family":"Xu","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Berahovich","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lijun","family":"Wu","sequence":"additional","affiliation":[{"name":"Promab Biotechnologies, 2600 Hilltop Drive, Richmond, CA 94806, USA"},{"name":"Laboratory for Critical Quality Attributes of Cell Therapy Products, Forevertek Biotechnology, Janshan Road, Changsha Hi-Tech Industrial Development Zone, Changsha 410205, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1002\/1873-3468.14270","article-title":"CAR-Ts: New perspectives in cancer therapy","volume":"596","author":"Abrantes","year":"2022","journal-title":"FEBS Lett."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1038\/s41408-021-00459-7","article-title":"CAR-T cell therapy: Current limitations and potential strategies","volume":"11","author":"Sterner","year":"2021","journal-title":"Blood Cancer J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3899","DOI":"10.1158\/1078-0432.CCR-14-0721","article-title":"CARTs on the road for myeloma","volume":"20","author":"Maus","year":"2014","journal-title":"Clin. 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