Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity
Adeno-associated virus (AAV) vectors have been successfully used in patients with bleeding disorders and blindness. For human liver targeting, two major factors restrict effective AAV transduction after systemic administration of AAV vectors: human hepatocyte tropism and neutralizing antibodies (Nab...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S232905012030125X |
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Article |
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DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Xiaolei Pei Wenwei Shao Allene Xing Charles Askew Xiaojing Chen Caibin Cui Yasmina L. Abajas David A. Gerber Elizabeth P. Merricks Timothy C. Nichols Wuping Li R. Jude Samulski Chengwen Li |
| spellingShingle |
Xiaolei Pei Wenwei Shao Allene Xing Charles Askew Xiaojing Chen Caibin Cui Yasmina L. Abajas David A. Gerber Elizabeth P. Merricks Timothy C. Nichols Wuping Li R. Jude Samulski Chengwen Li Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity Molecular Therapy: Methods & Clinical Development AAV human hepatocyte tropism Nabs chimeric mice |
| author_facet |
Xiaolei Pei Wenwei Shao Allene Xing Charles Askew Xiaojing Chen Caibin Cui Yasmina L. Abajas David A. Gerber Elizabeth P. Merricks Timothy C. Nichols Wuping Li R. Jude Samulski Chengwen Li |
| author_sort |
Xiaolei Pei |
| title |
Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity |
| title_short |
Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity |
| title_full |
Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity |
| title_fullStr |
Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity |
| title_full_unstemmed |
Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape Capacity |
| title_sort |
development of aav variants with human hepatocyte tropism and neutralizing antibody escape capacity |
| publisher |
Elsevier |
| series |
Molecular Therapy: Methods & Clinical Development |
| issn |
2329-0501 |
| publishDate |
2020-09-01 |
| description |
Adeno-associated virus (AAV) vectors have been successfully used in patients with bleeding disorders and blindness. For human liver targeting, two major factors restrict effective AAV transduction after systemic administration of AAV vectors: human hepatocyte tropism and neutralizing antibodies (Nabs). In this study, we attempted to isolate AAV variants with the ability to transduce human hepatocytes and escape Nabs using a directed evolution approach in vivo. After four cycles of selection, 14 AAV capsid mutants were identified from a capsid shuffling library selected in the presence of human Intravenous Immunoglobulin (IVIG) and isolated from human hepatocytes xenografted into chimeric mice. AAV neutralization assays using IVIG showed that most of the mutants showed the Nab escape pattern in a manner similar to that of AAV8 or AAV9 and better than that of other AAV serotypes. Different mutants displayed varying capacities to escape Nab activity from individual serum samples collected from healthy subjects or hemophilia patients. The mutant AAV LP2-10 was found in 12 colonies out of 25, which was composed of capsids from AAV serotypes 2, 6, 8, and 9, with VP3 subunits derived from AAV8 swapped with AAV6 from residues 261 to 272. The mutant AAV LP2-10 manifested a higher ability than that of other serotypes to escape Nabs in IVIG and most human serum samples. After injection of AAV vectors encoding a self-complementary GFP cassette into chimeric mice, LP2-10 transduced human hepatocytes with efficiency similar to that of AAV8. In summary, AAV mutants can be isolated in humanized mice with both human hepatocyte tropism and the ability to evade Nab activity. |
| topic |
AAV human hepatocyte tropism Nabs chimeric mice |
| url |
http://www.sciencedirect.com/science/article/pii/S232905012030125X |
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doaj-ffd1eccee8e6456aa953cf4176aa8c2d2020-11-25T02:54:23ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-09-0118259268Development of AAV Variants with Human Hepatocyte Tropism and Neutralizing Antibody Escape CapacityXiaolei Pei0Wenwei Shao1Allene Xing2Charles Askew3Xiaojing Chen4Caibin Cui5Yasmina L. Abajas6David A. Gerber7Elizabeth P. Merricks8Timothy C. Nichols9Wuping Li10R. Jude Samulski11Chengwen Li12State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China; Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAGene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAGene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAGene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAGene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USADepartment of Surgery, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USADepartment of Pediatrics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USADepartment of Surgery, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USADepartment of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USADepartment of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAInstitute of Pathogen Biology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, ChinaGene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USAGene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Pediatrics, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Carolina Institute for Developmental Disabilities, University of North Carolina at Chapel Hill, NC 27599, USA; Corresponding author: Chengwen Li, Gene Therapy Center, 7007 Thurston-Bowles, CB 7352, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.Adeno-associated virus (AAV) vectors have been successfully used in patients with bleeding disorders and blindness. For human liver targeting, two major factors restrict effective AAV transduction after systemic administration of AAV vectors: human hepatocyte tropism and neutralizing antibodies (Nabs). In this study, we attempted to isolate AAV variants with the ability to transduce human hepatocytes and escape Nabs using a directed evolution approach in vivo. After four cycles of selection, 14 AAV capsid mutants were identified from a capsid shuffling library selected in the presence of human Intravenous Immunoglobulin (IVIG) and isolated from human hepatocytes xenografted into chimeric mice. AAV neutralization assays using IVIG showed that most of the mutants showed the Nab escape pattern in a manner similar to that of AAV8 or AAV9 and better than that of other AAV serotypes. Different mutants displayed varying capacities to escape Nab activity from individual serum samples collected from healthy subjects or hemophilia patients. The mutant AAV LP2-10 was found in 12 colonies out of 25, which was composed of capsids from AAV serotypes 2, 6, 8, and 9, with VP3 subunits derived from AAV8 swapped with AAV6 from residues 261 to 272. The mutant AAV LP2-10 manifested a higher ability than that of other serotypes to escape Nabs in IVIG and most human serum samples. After injection of AAV vectors encoding a self-complementary GFP cassette into chimeric mice, LP2-10 transduced human hepatocytes with efficiency similar to that of AAV8. In summary, AAV mutants can be isolated in humanized mice with both human hepatocyte tropism and the ability to evade Nab activity.http://www.sciencedirect.com/science/article/pii/S232905012030125XAAVhuman hepatocytetropismNabschimeric mice |