Engineered exosome-mediated messenger RNA and single-chain variable fragment delivery for human chimeric antigen receptor T-cell engineering |
| |
| Institution: | 1. State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China;2. Department of Obstetrics and Gynecology, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China;1. Department of Cell Biology, Zunyi Medical University, Zunyi, Guizhou Province, China;2. Department of Orthopedic Surgery, Xinhua Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai, China;1. Chronic Disease Research Center, Medical College, Dalian University, Dalian, Liaoning, China;2. Engineering Technology Research Center for the Utilization of Functional Components of Organic Natural Products, Dalian University, Dalian, Liaoning, China;1. Department of Laboratory Medicine, Seoul St. Mary''s Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea;2. Department of Internal Medicine, Seoul St. Mary''s Hospital, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea;3. Research & Development Institute of In Vitro Diagnostic Medical Device of Catholic University of Korea, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea;4. Catholic Genetic Laboratory Center, College of Medicine, Seoul St. Mary''s Hospital, The Catholic University of Korea, Seoul, Republic of Korea;1. Pediatric Hematology-Oncology Fellow, Memorial Sloan Kettering Cancer Center, and Department of Pediatrics, Weill Cornell Medical College of Cornell University, New York, New York, USA;2. Center for Cancer and Immunology Research, CETI, Children''s National Hospital, Washington, District of Columbia, USA;3. Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Department of Pediatrics, Stanford University, Stanford, California, USA;4. Department of Bone Marrow Transplantation and Cellular Therapy, St Jude Children''s Research Hospital, Memphis, Tennessee, USA;5. Pediatric Blood and Marrow Transplantation Division and Pele Pequeno Principe Research Institute, Hospital Pequeno Principe, Curitiba, Brazil;6. Universite de Montreal and Maisonneuve Rosemont Hospital, Montreal, Quebec, Canada;7. Bone Marrow Failure and MDS Program, John Hopkins Medicine Baltimore, Maryland, USA;8. CRISPR Therapeutics Cambridge, Massachusetts, USA;9. Stem Cell Transplantation and Cellular Therapies, Memorial Sloan Kettering Cancer Center, New York, New York, USA;10. Department of Pediatrics, Division of Stem Cell Transplantation and Regenerative Medicine, Redwood City, California, USA;11. Department of Haematology, Fiona Stanley Hospital, Perth, Western Australia, Australia;12. IRCCS Ospedale San Raffaele, Segrate, Milan, Italy;13. Center for Cellular Immunotherapies, University of Pennsylvania, Philadelphia, Pennsylvania, USA;14. Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, UK;15. Stem Cell Transplantation and Cellular Therapies, Memorial Sloan Kettering Cancer Center, and Department of Pediatrics, Weill Cornell Medical College of Cornell University, New York, New York, USA;16. Dana-Farber/Boston Children''s Cancer and Blood Disorders Center, Boston, Massachusetts USA;1. Pediatrics, Nationwide Children''s Hospital, Columbus, Ohio, USA;2. Center for Regenerative Medicine, Mayo Clinic Florida, Jacksonville, Florida, USA;3. Department of Neurosurgery, Duke University School of Medicine, Durham, North Carolina, USA;4. Center for Cellular Immunotherapies, Philadelphia, Pennsylvania, USA;5. Steno Diabetes Center Copenhagen, Copenhagen, Denmark;6. Faculty of Medicine and Health (FML), FML Medical Research Institutes, University of Sydney, Sydney Australia;7. Obsidian Therapeutics, Cambridge, Massachusetts, USA;1. Department of Cardiology, Rostock University Medical Center, Rostock, Germany;2. Department of Cardiac Surgery, Rostock University Medical Center, Rostock, Germany;3. Department of Life, Light and Matter, University of Rostock, Rostock, Germany;4. Institute of Medical Informatics and Biometry, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany |
| |
| Abstract: | Background aimsMost current chimeric antigen receptor (CAR) T cells are generated by viral transduction, which induces persistent expression of CARs and may cause serious undesirable effects. Messenger RNA (mRNA)-based approaches in manufacturing CAR T cells are being developed to overcome these challenges. However, the most common method of delivering mRNA to T cells is electroporation, which can be toxic to cells.MethodsThe authors designed and engineered an exosome delivery platform using the bacteriophage MS2 system in combination with the highly expressed protein lysosome-associated membrane protein 2 isoform B on exosomes.ResultsThe authors’ delivery platform achieved specific loading and delivery of mRNA into target cells and achieved expression of specific proteins, and anti-CD3/CD28 single-chain variable fragments (scFvs) expressed outside the exosomal membrane effectively activated primary T cells in a similar way to commercial magnetic beads.ConclusionsThe delivery of CAR mRNA and anti-CD3/CD28 scFvs via designed exosomes can be used for ex vivo production of CAR T cells with cancer cell killing capacity. The authors’ results indicate the potential applications of the engineered exosome delivery platform for direct conversion of primary T cells to CAR T cells while providing a novel strategy for producing CAR T cells in vivo. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
