{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T02:34:51Z","timestamp":1792722891828,"version":"4.5.0"},"reference-count":49,"publisher":"National Academy of Sciences","issue":"29","license":[{"start":{"date-parts":[[2025,7,14]],"date-time":"2025-07-14T00:00:00Z","timestamp":1752451200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"MOST | National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82373287 82172762"],"award-info":[{"award-number":["82373287 82172762"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"MOST | National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["82222067 82404531"],"award-info":[{"award-number":["82222067 82404531"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2025,7,22]]},"abstract":"<jats:p>Glioblastoma multiforme (GBM) is one of the most aggressive intracranial tumors for which there is no effective treatment. Chimeric antigen receptor macrophage (CAR-M) therapies have demonstrated impressive therapeutic efficacy in solid tumors; however, the cost and rigor associated with manufacturing engineered macrophages ex vivo can be prohibitive. Here, we utilized enucleated mesenchymal stem cells (MSCs) as vehicles for the targeted delivery of CAR-encoding plasmid to reprogram glioma-associated microglia\/macrophages (GAM), thereby achieving CAR-M preparation in vivo. Specifically, we observed that the enucleated cells retained the key organelle function and membrane integrity, and actively homed to glioma tissue. Interestingly, enucleated MSCs underwent intrinsic apoptosis due to the absence of the nucleus, which subsequently triggered macrophage-specific endocytosis, thereby achieving precise delivery of CAR-plasmids to GAM. Compared with lipid nanoparticles, this strategy specifically generated sufficient numbers of CAR-M in glioma situ to achieve GBM therapy. Moreover, this process altered the immune cell profiles within the tumor by increasing cytotoxic T cells and M1-like macrophages with antitumor activity. When combined with CD47-blocking therapies, tumor growth was completely suppressed in the GBM orthotopic mouse model, resulting in a 90-d survival rate of 83%. Collectively, our strategy provides a viable platform technology for CAR-M generation in vivo, which is expected to provide an approach for GBM therapy.<\/jats:p>","DOI":"10.1073\/pnas.2426724122","type":"journal-article","created":{"date-parts":[[2025,7,14]],"date-time":"2025-07-14T19:01:31Z","timestamp":1752519691000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":36,"title":["In vivo generation of CAR macrophages via the enucleated mesenchymal stem cell delivery system for glioblastoma therapy"],"prefix":"10.1073","volume":"122","author":[{"given":"Lei","family":"Zhou","sequence":"first","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qingying","family":"Song","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengnian","family":"Cao","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dayu","family":"Xue","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiwen","family":"Sun","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Meihua","family":"Mao","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinling","family":"Li","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhenzhong","family":"Zhang","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]},{"name":"Key Laboratory of Targeting Therapy and Diagnosis for Critical Diseases","place":["Zhengzhou, China"]},{"name":"School of Pharmaceutical Sciences, State Key Laboratory of Antiviral Drugs, Pingyuan Laboratory, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9404-050X","authenticated-orcid":false,"given":"Junjie","family":"Liu","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1531-7265","authenticated-orcid":false,"given":"Jinjin","family":"Shi","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences, Zhengzhou University","place":["Zhengzhou, China"]},{"name":"Key Laboratory of Targeting Therapy and Diagnosis for Critical Diseases","place":["Zhengzhou, China"]},{"name":"School of Pharmaceutical Sciences, State Key Laboratory of Antiviral Drugs, Pingyuan Laboratory, Zhengzhou University","place":["Zhengzhou, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2025,7,14]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-022-00894-3"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s43018-021-00238-0"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-022-31799-y"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41565-022-01189-y"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0462-y"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.ebiom.2022.103873"},{"key":"e_1_3_4_7_2","doi-asserted-by":"crossref","unstructured":"K. A. Reiss A phase 1 first-in-human (FIH) study of adenovirally transduced autologous macrophages engineered to contain an anti-HER2 chimeric antigen receptor (CAR) in participants with HER2 overexpressing solid tumors. J. Clin. 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Zhou Transcriptome sequencing of BMDM from mus musculus (males) treated with enucleated MSC delivery system. NCBI. https:\/\/www.ncbi.nlm.nih.gov\/bioproject\/PRJNA1194746. Deposited 6 December 2024."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2426724122","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,7,14]],"date-time":"2025-07-14T19:02:42Z","timestamp":1752519762000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2426724122"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7,14]]},"references-count":49,"journal-issue":{"issue":"29","published-print":{"date-parts":[[2025,7,22]]}},"alternative-id":["10.1073\/pnas.2426724122"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2426724122","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7,14]]},"assertion":[{"value":"2024-12-25","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-05-13","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-14","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2426724122"}}