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Mol Ther. 2016 Feb;24(1):53-65. doi: 10.1038/mt.2015.134. Epub 2015 Jul 29.

Development of Patient-specific AAV Vectors After Neutralizing Antibody Selection for Enhanced Muscle Gene Transfer.

Author information

1
Gene Therapy Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
2
Department of Pediatrics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
3
China National Academy of Nanotechnology & Engineering, Tianjin, China.
4
Institute of Pathogen Biology, Chinese Academy of Medical Sciences, Beijing, China.
5
Department of Biochemistry and Molecular Biology, Center for Structural Biology, McKnight Brain Institute, University of Florida, Gainesville, Florida, USA.
6
Asklepios BioPharmaceutical Inc., Chapel Hill, North Carolina, USA.
7
Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.
8
Department of Pharmacology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, USA.

Abstract

A major hindrance in gene therapy trials with adeno-associated virus (AAV) vectors is the presence of neutralizing antibodies (NAbs) that inhibit AAV transduction. In this study, we used directed evolution techniques in vitro and in mouse muscle to select novel NAb escape AAV chimeric capsid mutants in the presence of individual patient serum. AAV mutants isolated in vitro escaped broad patient-specific NAb activity but had poor transduction ability in vivo. AAV mutants isolated in vivo had enhanced NAb evasion from cognate serum and had high muscle transduction ability. More importantly, structural modeling identified a 100 amino acid motif from AAV6 in variable region (VR) III that confers this enhanced muscle tropism. In addition, a predominantly AAV8 capsid beta barrel template with a specific preference for AAV1/AAV9 in VR VII located at threefold symmetry axis facilitates NAb escape. Our data strongly support that chimeric AAV capsids composed of modular and nonoverlapping domains from various serotypes are capable of evading patient-specific NAbs and have enhanced muscle transduction.

PMID:
26220272
PMCID:
PMC4754536
DOI:
10.1038/mt.2015.134
[Indexed for MEDLINE]
Free PMC Article

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