Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties
Extracellular vesicle-associated adeno-associated virus vectors (EV-AAVs) are generated during production in 293 cells. EV-AAV provides desirable gene delivery traits such as greater resistance to antibody neutralization and increased transduction of organs in vivo compared with conventional AAV. De...
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Elsevier
2025-03-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050125000026 |
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| author | Ming Cheng Demitri de la Cruz Adam V. Crain Paula Espinoza Carrie Ng Zachary C. Elmore Aravind Asokan Casey A. Maguire |
| author_facet | Ming Cheng Demitri de la Cruz Adam V. Crain Paula Espinoza Carrie Ng Zachary C. Elmore Aravind Asokan Casey A. Maguire |
| author_sort | Ming Cheng |
| collection | DOAJ |
| description | Extracellular vesicle-associated adeno-associated virus vectors (EV-AAVs) are generated during production in 293 cells. EV-AAV provides desirable gene delivery traits such as greater resistance to antibody neutralization and increased transduction of organs in vivo compared with conventional AAV. Despite these promising data, better characterization of EV-AAV is needed. We used density gradient ultracentrifugation to separate EV-AAV from free AAV to determine the yields and functional activity of EV-AAV. We found that the fraction of EV-AAV to conventional AAV in culture media from six AAV serotypes ranged from 0.5% to 12%. Next, we assessed whether intraluminal EV-AAV9 could mediate functional transduction of cells and observed that a portion of EV-AAV9 are intraluminal and mediated transduction of cultured cells in vitro and in vivo and evade antibodies compared with conventional AAV9. We tested whether trans-expression of membrane-associated accessory protein (MAAP) from AAV8 (MAAP8) or AAV9 (MAAP9) with AAV9 Cap/AAV9 MAAP null would alter yields of EV-AAV9. Trans-expression of MAAP8 or MAAP9 increased yields of EV-AAV9 compared with the cis-expression of AAV9 Cap/AAV9 MAAP. Finally, we found that the capsid was required for efficient transduction of cultured cells by EV-AAV. In sum, these data provide a foundation for the development of EV-AAV vectors. |
| format | Article |
| id | doaj-art-169cfefbba2041acb143632469358155 |
| institution | Kabale University |
| issn | 2329-0501 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
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| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj-art-169cfefbba2041acb1436324693581552025-02-08T05:00:33ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012025-03-01331101407Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive propertiesMing Cheng0Demitri de la Cruz1Adam V. Crain2Paula Espinoza3Carrie Ng4Zachary C. Elmore5Aravind Asokan6Casey A. Maguire7Department of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USADepartment of Surgery, Duke University School of Medicine, Durham, NC 27710, USADepartment of Surgery, Duke University School of Medicine, Durham, NC 27710, USA; Department of Molecular Genetics & Microbiology, Duke University School of Medicine, Durham, NC 27710, USA; Department of Biomedical Engineering, Duke University, Durham, NC 27710, USADepartment of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA; Molecular Neurogenetics Unit, Massachusetts General Hospital, Charlestown, MA 02129, USA; Harvard Medical School, Boston, MA 02115, USA; Corresponding author: Casey A. Maguire, Department of Neurology, Massachusetts General Hospital, Boston, MA 02114, USA.Extracellular vesicle-associated adeno-associated virus vectors (EV-AAVs) are generated during production in 293 cells. EV-AAV provides desirable gene delivery traits such as greater resistance to antibody neutralization and increased transduction of organs in vivo compared with conventional AAV. Despite these promising data, better characterization of EV-AAV is needed. We used density gradient ultracentrifugation to separate EV-AAV from free AAV to determine the yields and functional activity of EV-AAV. We found that the fraction of EV-AAV to conventional AAV in culture media from six AAV serotypes ranged from 0.5% to 12%. Next, we assessed whether intraluminal EV-AAV9 could mediate functional transduction of cells and observed that a portion of EV-AAV9 are intraluminal and mediated transduction of cultured cells in vitro and in vivo and evade antibodies compared with conventional AAV9. We tested whether trans-expression of membrane-associated accessory protein (MAAP) from AAV8 (MAAP8) or AAV9 (MAAP9) with AAV9 Cap/AAV9 MAAP null would alter yields of EV-AAV9. Trans-expression of MAAP8 or MAAP9 increased yields of EV-AAV9 compared with the cis-expression of AAV9 Cap/AAV9 MAAP. Finally, we found that the capsid was required for efficient transduction of cultured cells by EV-AAV. In sum, these data provide a foundation for the development of EV-AAV vectors.http://www.sciencedirect.com/science/article/pii/S2329050125000026adeno-associated virus vectorsAAV vectorsextracellular vesiclesEVsexosomesgene delivery |
| spellingShingle | Ming Cheng Demitri de la Cruz Adam V. Crain Paula Espinoza Carrie Ng Zachary C. Elmore Aravind Asokan Casey A. Maguire Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties Molecular Therapy: Methods & Clinical Development adeno-associated virus vectors AAV vectors extracellular vesicles EVs exosomes gene delivery |
| title | Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties |
| title_full | Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties |
| title_fullStr | Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties |
| title_full_unstemmed | Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties |
| title_short | Probing aspects of extracellular vesicle associated AAV allows increased vector yield and insight into its transduction and immune-evasive properties |
| title_sort | probing aspects of extracellular vesicle associated aav allows increased vector yield and insight into its transduction and immune evasive properties |
| topic | adeno-associated virus vectors AAV vectors extracellular vesicles EVs exosomes gene delivery |
| url | http://www.sciencedirect.com/science/article/pii/S2329050125000026 |
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