Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model
A limitation for recombinant adeno-associated virus (rAAV)-mediated gene transfer into the central nervous system (CNS) is the low penetration of vectors across the human blood-brain barrier (BBB). High doses of intravenously delivered vector are required to reach the CNS, which has resulted in vary...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050122001279 |
| _version_ | 1818019947226333184 |
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| author | Ren Song Katja Pekrun Themasap A. Khan Feijie Zhang Sergiu P. Paşca Mark A. Kay |
| author_facet | Ren Song Katja Pekrun Themasap A. Khan Feijie Zhang Sergiu P. Paşca Mark A. Kay |
| author_sort | Ren Song |
| collection | DOAJ |
| description | A limitation for recombinant adeno-associated virus (rAAV)-mediated gene transfer into the central nervous system (CNS) is the low penetration of vectors across the human blood-brain barrier (BBB). High doses of intravenously delivered vector are required to reach the CNS, which has resulted in varying adverse effects. Moreover, selective transduction of various cell types might be important depending on the disorder being treated. To enhance BBB penetration and improve CNS cell selectivity, we screened an AAV capsid-shuffled library using an in vitro transwell BBB system with separate layers of human endothelial cells, primary astrocytes and/or human induced pluripotent stem cell-derived cortical neurons. After multiple passages through the transwell, we identified chimeric AAV capsids with enhanced penetration and improved transduction of astrocytes and/or neurons compared with wild-type capsids. We identified the amino acids (aa) from regions 451–470 of AAV2 associated with the capsids selected for neurons, and a combination of aa from regions 413–496 of AAV-rh10 and 538–598 of AAV3B/LK03 associated with capsids selected for astrocytes. A small interfering RNA screen identified several genes that affect transcytosis of AAV across the BBB. Our work supports the use of a human transwell system for selecting enhanced AAV capsids targeting the CNS and may allow for unraveling the underlying molecular mechanisms of BBB penetration. |
| first_indexed | 2024-04-14T07:58:34Z |
| format | Article |
| id | doaj.art-86db400fe64f437aa8bcb31f674ac266 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-04-14T07:58:34Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-86db400fe64f437aa8bcb31f674ac2662022-12-22T02:04:57ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012022-12-01277388Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell modelRen Song0Katja Pekrun1Themasap A. Khan2Feijie Zhang3Sergiu P. Paşca4Mark A. Kay5Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305, USADepartments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305, USADepartment of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Brain Organogenesis, Wu Tsai Neuroscience Institute and Bio-X, Stanford University, CA 94305, USADepartments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305, USADepartment of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, CA 94305, USA; Stanford Brain Organogenesis, Wu Tsai Neuroscience Institute and Bio-X, Stanford University, CA 94305, USADepartments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA; Corresponding author Mark A. Kay, Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.A limitation for recombinant adeno-associated virus (rAAV)-mediated gene transfer into the central nervous system (CNS) is the low penetration of vectors across the human blood-brain barrier (BBB). High doses of intravenously delivered vector are required to reach the CNS, which has resulted in varying adverse effects. Moreover, selective transduction of various cell types might be important depending on the disorder being treated. To enhance BBB penetration and improve CNS cell selectivity, we screened an AAV capsid-shuffled library using an in vitro transwell BBB system with separate layers of human endothelial cells, primary astrocytes and/or human induced pluripotent stem cell-derived cortical neurons. After multiple passages through the transwell, we identified chimeric AAV capsids with enhanced penetration and improved transduction of astrocytes and/or neurons compared with wild-type capsids. We identified the amino acids (aa) from regions 451–470 of AAV2 associated with the capsids selected for neurons, and a combination of aa from regions 413–496 of AAV-rh10 and 538–598 of AAV3B/LK03 associated with capsids selected for astrocytes. A small interfering RNA screen identified several genes that affect transcytosis of AAV across the BBB. Our work supports the use of a human transwell system for selecting enhanced AAV capsids targeting the CNS and may allow for unraveling the underlying molecular mechanisms of BBB penetration.http://www.sciencedirect.com/science/article/pii/S2329050122001279blood brain barrierCNS capsidstranswellhuman BBBcentral nervous systemchimeric AAV vectors |
| spellingShingle | Ren Song Katja Pekrun Themasap A. Khan Feijie Zhang Sergiu P. Paşca Mark A. Kay Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model Molecular Therapy: Methods & Clinical Development blood brain barrier CNS capsids transwell human BBB central nervous system chimeric AAV vectors |
| title | Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model |
| title_full | Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model |
| title_fullStr | Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model |
| title_full_unstemmed | Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model |
| title_short | Selection of rAAV vectors that cross the human blood-brain barrier and target the central nervous system using a transwell model |
| title_sort | selection of raav vectors that cross the human blood brain barrier and target the central nervous system using a transwell model |
| topic | blood brain barrier CNS capsids transwell human BBB central nervous system chimeric AAV vectors |
| url | http://www.sciencedirect.com/science/article/pii/S2329050122001279 |
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