Journal
CYTOTHERAPY
Volume 15, Issue 8, Pages 986-998Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.jcyt.2013.04.003
Keywords
AAV vectors; gene expression; gene transfer; hematopoietic stem cells
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Funding
- Fanconi Anemia Research Fund, Inc
- Natural Science Foundation of China (NSFC) [30971299]
- Public Health Service from the National Institutes of Health [R01 HL-087285, P30 CA-33572, R01 HL-065770, HL-076901, P01 DK-058327, R01 HL-097088]
- Children's Miracle Network
- Department of Biotechnology, Government of India
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Background aims. Although recombinant adeno-associated virus serotype 2 (AAV2) vectors have gained attention because of their safety and efficacy in numerous phase I/II clinical trials, their transduction efficiency in hematopoietic stem cells (HSCs) has been reported to be low. Only a few additional AAV serotype vectors have been evaluated, and comparative analyses of their transduction efficiency in HSCs from different species have not been performed. Methods. We evaluated the transduction efficiency of all available AAV serotype vectors (AAV1 through AAV10) in primary mouse, cynomolgus monkey and human HSCs. The transduction efficiency of the optimized AAV vectors was also evaluated in human HSCs in a murine xenograft model in vivo. Results. We observed that although there are only six amino acid differences between AAV1 and AAV6, AAV1, but not AAV6, transduced mouse HSCs well, whereas AAV6, but not AAV1, transduced human HSCs well. None of the 10 serotypes transduced cynomolgus monkey HSCs in vitro. We also evaluated the transduction efficiency of AAV6 vectors containing mutations in surface-exposed tyrosine residues. We observed that tyrosine (Y) to phenylalanine (F) point mutations in residues 445, 705 and 731 led to a significant increase in transgene expression in human HSCs in vitro and in a mouse xenograft model in vivo. Conclusions. These studies suggest that the tyrosine-mutant AAV6 serotype vectors are the most promising vectors for transducing human HSCs and that it is possible to increase further the transduction efficiency of these vectors for their potential use in HSC-based gene therapy in humans.
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