{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T12:50:43Z","timestamp":1788958243311,"version":"build-2803163510"},"reference-count":40,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,16]],"date-time":"2024-05-16T00:00:00Z","timestamp":1715817600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Hunan Provincial Natural Science Foundation of China","award":["2021JJ30801"],"award-info":[{"award-number":["2021JJ30801"]}]},{"name":"Hunan Provincial Natural Science Foundation of China","award":["2021zzts0565"],"award-info":[{"award-number":["2021zzts0565"]}]},{"name":"Hunan Provincial Natural Science Foundation of China","award":["kq2202077"],"award-info":[{"award-number":["kq2202077"]}]},{"name":"the Postgraduate Freedom Exploration Project of Central South University","award":["2021JJ30801"],"award-info":[{"award-number":["2021JJ30801"]}]},{"name":"the Postgraduate Freedom Exploration Project of Central South University","award":["2021zzts0565"],"award-info":[{"award-number":["2021zzts0565"]}]},{"name":"the Postgraduate Freedom Exploration Project of Central South University","award":["kq2202077"],"award-info":[{"award-number":["kq2202077"]}]},{"name":"the Natural Science Foundation of Changsha","award":["2021JJ30801"],"award-info":[{"award-number":["2021JJ30801"]}]},{"name":"the Natural Science Foundation of Changsha","award":["2021zzts0565"],"award-info":[{"award-number":["2021zzts0565"]}]},{"name":"the Natural Science Foundation of Changsha","award":["kq2202077"],"award-info":[{"award-number":["kq2202077"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biomolecules"],"abstract":"<jats:p>Small extracellular vesicles (sEVs) have emerged as promising therapeutic agents and drug delivery vehicles. Targeted modification of sEVs and their contents using genetic modification strategies is one of the most popular methods. This study investigated the effects of p53 fusion with arrestin domain-containing protein 1 (ARRDC1) and CD63 on the generation of sEVs, p53 loading efficiency, and therapeutic efficacy. Overexpression of either ARRDC1\u2013p53 (ARP) or CD63\u2013p53 (CDP) significantly elevated p53 mRNA and protein levels. The incorporation of ARRDC1 and CD63 significantly enhanced HEK293T-sEV biogenesis, evidenced by significant increases in sEV-associated proteins TSG101 and LAMP1, resulting in a boost in sEV production. Importantly, fusion with ARRDC1 or CD63 substantially increased the efficiency of loading both p53 fusion proteins and its mRNA into sEVs. sEVs equipped with ARP or CDP significantly enhanced the enrichment of p53 fusion proteins and mRNA in p53-null H1299 cells, resulting in a marked increase in apoptosis and a reduction in cell proliferation, with ARP-sEVs demonstrating greater effectiveness than CDP-sEVs. These findings underscore the enhanced functionality of ARRDC1- and CD63-modified sEVs, emphasizing the potential of genetic modifications in sEV-based therapies for targeted cancer treatment.<\/jats:p>","DOI":"10.3390\/biom14050591","type":"journal-article","created":{"date-parts":[[2024,5,16]],"date-time":"2024-05-16T11:26:17Z","timestamp":1715858777000},"page":"591","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Fusion with ARRDC1 or CD63: A Strategy to Enhance p53 Loading into Extracellular Vesicles for Tumor Suppression"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3965-846X","authenticated-orcid":false,"given":"Min","family":"Liu","sequence":"first","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Zhang","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianfeng","family":"He","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"},{"name":"Department of Cell Biology, School of Life Sciences, Central South University, Changsha 410013, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanxi","family":"Liu","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhexuan","family":"Li","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4245-2879","authenticated-orcid":false,"given":"Yiti","family":"Zhang","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ao","family":"Gu","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingri","family":"Zhao","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1558-1448","authenticated-orcid":false,"given":"Mujun","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, School of Life Sciences, Central South University, Changsha 410013, China"},{"name":"Hunan Key Laboratory of Basic and Applied Hematology, Central South University, Changsha 410078, China"},{"name":"Hunan Key Laboratory of Animal Models for Human Diseases, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2633-7096","authenticated-orcid":false,"given":"Xionghao","family":"Liu","sequence":"additional","affiliation":[{"name":"Center for Medical Genetics & Hunan Key Laboratory of Medical Genetics, School of Life Sciences, Central South University, Changsha 410078, China"},{"name":"Hunan Key Laboratory of Basic and Applied Hematology, Central South University, Changsha 410078, China"},{"name":"Hunan Key Laboratory of Animal Models for Human Diseases, Central South University, Changsha 410078, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/nrm.2017.125","article-title":"Shedding Light on the Cell Biology of Extracellular Vesicles","volume":"19","author":"Raposo","year":"2018","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1038\/s41580-020-0251-y","article-title":"RNA Delivery by Extracellular Vesicles in Mammalian Cells and Its Applications","volume":"21","author":"Breyne","year":"2020","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1535750","DOI":"10.1080\/20013078.2018.1535750","article-title":"Minimal Information for Studies of Extracellular Vesicles 2018 (MISEV2018): A Position Statement of the International Society for Extracellular Vesicles and Update of the MISEV2014 Guidelines","volume":"7","author":"Witwer","year":"2018","journal-title":"J. Extracell. Vesicles"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"22519","DOI":"10.1038\/srep22519","article-title":"Cells Release Subpopulations of Exosomes with Distinct Molecular and Biological Properties","volume":"6","author":"Willms","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"4146","DOI":"10.1073\/pnas.1200448109","article-title":"Formation and Release of Arrestin Domain-Containing Protein 1-Mediated Microvesicles (ARMMs) at Plasma Membrane by Recruitment of TSG101 Protein","volume":"109","author":"Nabhan","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"498","DOI":"10.1038\/nature22341","article-title":"Exosomes Facilitate Therapeutic Targeting of Oncogenic KRAS in Pancreatic Cancer","volume":"546","author":"Kamerkar","year":"2017","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e12144","DOI":"10.1002\/jev2.12144","article-title":"A Brief History of Nearly EV-Erything\u2014The Rise and Rise of Extracellular Vesicles","volume":"10","author":"Couch","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"237","DOI":"10.7150\/thno.21945","article-title":"Exosome Theranostics: Biology and Translational Medicine","volume":"8","author":"He","year":"2018","journal-title":"Theranostics"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e10250","DOI":"10.7554\/eLife.10250","article-title":"Tumor Microenvironment Derived Exosomes Pleiotropically Modulate Cancer Cell Metabolism","volume":"5","author":"Zhao","year":"2016","journal-title":"Elife"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1769","DOI":"10.1038\/mt.2011.164","article-title":"Treatment of Brain Inflammatory Diseases by Delivering Exosome Encapsulated Anti-Inflammatory Drugs from the Nasal Region to the Brain","volume":"19","author":"Zhuang","year":"2011","journal-title":"Mol. Ther."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1038\/nmeth.4579","article-title":"EVIR: Chimeric Receptors That Enhance Dendritic Cell Cross-Dressing with Tumor Antigens","volume":"15","author":"Squadrito","year":"2018","journal-title":"Nat. Methods"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Gangadaran, P., and Ahn, B.-C. (2020). Extracellular Vesicle-and Extracellular Vesicle Mimetics-Based Drug Delivery Systems: New Perspectives, Challenges, and Clinical Developments. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12050442"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"F906","DOI":"10.1152\/ajprenal.00178.2017","article-title":"HIF-1-Mediated Production of Exosomes during Hypoxia Is Protective in Renal Tubular Cells","volume":"313","author":"Zhang","year":"2017","journal-title":"Am. J. Physiol. Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1038\/nbt.1807","article-title":"Delivery of SiRNA to the Mouse Brain by Systemic Injection of Targeted Exosomes","volume":"29","author":"Seow","year":"2011","journal-title":"Nat. Biotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e02251-16","DOI":"10.1128\/JVI.02251-16","article-title":"CD63 Regulates Epstein-Barr Virus LMP1 Exosomal Packaging, Enhancement of Vesicle Production, and Noncanonical NF-\u039aB Signaling","volume":"91","author":"Hurwitz","year":"2017","journal-title":"J. Virol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e12076","DOI":"10.1002\/jev2.12076","article-title":"Engineered Extracellular Vesicles as Versatile Ribonucleoprotein Delivery Vehicles for Efficient and Safe CRISPR Genome Editing","volume":"10","author":"Yao","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1038\/s41467-018-03390-x","article-title":"ARMMs as a Versatile Platform for Intracellular Delivery of Macromolecules","volume":"9","author":"Wang","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1002\/bit.28322","article-title":"Production Cell Analysis and Compound-Based Boosting of Small Extracellular Vesicle Secretion Using a Generic and Scalable Production Platform","volume":"120","author":"Keysberg","year":"2022","journal-title":"Biotechnol. Bioeng."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Song, Z., Mao, J., Barrero, R.A., Wang, P., Zhang, F., and Wang, T. (2020). Development of a CD63 Aptamer for Efficient Cancer Immunochemistry and Immunoaffinity-Based Exosome Isolation. Molecules, 25.","DOI":"10.3390\/molecules25235585"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"22614","DOI":"10.3402\/jev.v2i0.22614","article-title":"Therapeutic MSC Exosomes Are Derived from Lipid Raft Microdomains in the Plasma Membrane","volume":"2","author":"Tan","year":"2013","journal-title":"J. Extracell. Vesicles"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Zhang, R., Bu, T., Cao, R., Li, Z., Wang, C., Huang, B., Wei, M., Yuan, L., and Yang, G. (2022). An Optimized Exosome Production Strategy for Enhanced Yield While without Sacrificing Cargo Loading Efficiency. J. Nanobiotechnology, 20.","DOI":"10.1186\/s12951-022-01668-3"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1080\/10717544.2020.1748758","article-title":"Prospects and Challenges of Extracellular Vesicle-Based Drug Delivery System: Considering Cell Source","volume":"27","author":"Meng","year":"2020","journal-title":"Drug Deliv."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1098","DOI":"10.1080\/07388551.2020.1805406","article-title":"Improvement, Scaling-up, and Downstream Analysis of Exosome Production","volume":"40","author":"Jafari","year":"2020","journal-title":"Crit. Rev. Biotechnol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Liang, G., Zhu, Y., Ali, D.J., Tian, T., Xu, H., Si, K., Sun, B., Chen, B., and Xiao, Z. (2020). Engineered Exosomes for Targeted Co-Delivery of MiR-21 Inhibitor and Chemotherapeutics to Reverse Drug Resistance in Colon Cancer. J. Nanobiotechnol., 18.","DOI":"10.1186\/s12951-019-0563-2"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1800222","DOI":"10.1080\/20013078.2020.1800222","article-title":"Quantification of Extracellular Vesicles In Vitro and In Vivo Using Sensitive Bioluminescence Imaging","volume":"9","author":"Gupta","year":"2020","journal-title":"J. Extracell. Vesicles"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Wallis, B., Bowman, K.R., Lu, P., and Lim, C.S. (2023). The Challenges and Prospects of P53-Based Therapies in Ovarian Cancer. Biomolecules, 13.","DOI":"10.3390\/biom13010159"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"40791","DOI":"10.18632\/oncotarget.16584","article-title":"Enhanced Tumor Growth Inhibition by Mesenchymal Stem Cells Derived from IPSCs with Targeted Integration of Interleukin24 into RDNA Loci","volume":"8","author":"Liu","year":"2017","journal-title":"Oncotarget"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"163","DOI":"10.7150\/ijbs.53671","article-title":"Regulation of Exosome Production and Cargo Sorting","volume":"17","author":"Wei","year":"2020","journal-title":"Int. J. Biol. Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1038\/nature07961","article-title":"The ESCRT Machinery in Endosomal Sorting of Ubiquitylated Membrane Proteins","volume":"458","author":"Raiborg","year":"2009","journal-title":"Nature"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1038\/s41375-019-0493-x","article-title":"Targeting Histone Deacetylase 3 (HDAC3) in the Bone Marrow Microenvironment Inhibits Multiple Myeloma Proliferation by Modulating Exosomes and IL-6 Trans-Signaling","volume":"34","author":"Ho","year":"2020","journal-title":"Leukemia"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4389","DOI":"10.1038\/s41467-021-24384-2","article-title":"Specificities of Exosome versus Small Ectosome Secretion Revealed by Live Intracellular Tracking of CD63 and CD9","volume":"12","author":"Mathieu","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Nam, E., Lee, Y.-B., Moon, C., and Chang, K.-A. (2020). Serum Tau Proteins as Potential Biomarkers for the Assessment of Alzheimer\u2019s Disease Progression. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21145007"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1285","DOI":"10.1083\/jcb.200902147","article-title":"The Enterocyte Microvillus is a Vesicle-Generating Organelle","volume":"185","author":"McConnell","year":"2009","journal-title":"J. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1038\/ncb2502","article-title":"Syndecan\u2013Syntenin\u2013ALIX Regulates the Biogenesis of Exosomes","volume":"14","author":"Baietti","year":"2012","journal-title":"Nat. Cell Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1038\/s41467-017-00767-2","article-title":"Plasma Membrane-Derived Extracellular Microvesicles Mediate Non-Canonical Intercellular NOTCH Signaling","volume":"8","author":"Wang","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"512","DOI":"10.1016\/j.omtn.2020.06.013","article-title":"Therapeutic Effects of Simultaneous Delivery of Nerve Growth Factor MRNA and Protein via Exosomes on Cerebral Ischemia","volume":"21","author":"Yang","year":"2020","journal-title":"Mol. Ther.\u2014Nucleic Acids"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Li, F., Wu, J., Li, D., Hao, L., Li, Y., Yi, D., Yeung, K.W.K., Chen, D., Lu, W.W., and Pan, H. (2022). Engineering Stem Cells to Produce Exosomes with Enhanced Bone Regeneration Effects: An Alternative Strategy for Gene Therapy. J. Nanobiotechnology, 20.","DOI":"10.1186\/s12951-022-01347-3"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e12053","DOI":"10.1002\/jev2.12053","article-title":"V Mesenchymal Stem Cell Exosome Delivered Zinc Finger Protein Activation of Cystic Fibrosis Transmembrane Conductance Regulator","volume":"10","author":"Villamizar","year":"2021","journal-title":"J. Extracell. Vesicles"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2237","DOI":"10.1038\/s41418-019-0293-x","article-title":"Derepression of LOXL4 Inhibits Liver Cancer Growth by Reactivating Compromised P53","volume":"26","author":"Shao","year":"2019","journal-title":"Cell Death Differ."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1189\/jlb.3A1015-480R","article-title":"CD63 is Tightly Associated with Intracellular, Secretory Events Chaperoning Piecemeal Degranulation and Compound Exocytosis in Human Eosinophils","volume":"100","author":"Carmo","year":"2016","journal-title":"J. Leucoc. Biol."}],"container-title":["Biomolecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2218-273X\/14\/5\/591\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:43:36Z","timestamp":1760107416000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2218-273X\/14\/5\/591"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,16]]},"references-count":40,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["biom14050591"],"URL":"https:\/\/doi.org\/10.3390\/biom14050591","relation":{},"ISSN":["2218-273X"],"issn-type":[{"value":"2218-273X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,16]]}}}