Integration profile and safety of an adenovirus hybrid-vector utilizing hyperactive Sleeping Beauty transposase for somatic integration

Zhang, W.; Muck-Hausl, M.; Wang, J.; Sun, C.; Gebbing, M.; Miskey, C.; Ivics, Z.; Izsvak, Z.; Ehrhardt, A.

Integration profile and safety of an adenovirus hybrid-vector utilizing hyperactive Sleeping Beauty transposase for somatic integration

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Item Type:	Article
Title:	Integration profile and safety of an adenovirus hybrid-vector utilizing hyperactive Sleeping Beauty transposase for somatic integration
Creators Name:	Zhang, W., Muck-Hausl, M., Wang, J., Sun, C., Gebbing, M., Miskey, C., Ivics, Z., Izsvak, Z. and Ehrhardt, A.
Abstract:	We recently developed adenovirus/transposase hybrid-vectors utilizing the previously described hyperactive Sleeping Beauty (SB) transposase HSB5 for somatic integration and we could show stabilized transgene expression in mice and a canine model for hemophilia B. However, the safety profile of these hybrid-vectors with respect to vector dose and genotoxicity remains to be investigated. Herein, we evaluated this hybrid-vector system in C57Bl/6 mice with escalating vector dose settings. We found that in all mice which received the hyperactive SB transposase, transgene expression levels were stabilized in a dose-dependent manner and that the highest vector dose was accompanied by fatalities in mice. To analyze potential genotoxic side-effects due to somatic integration into host chromosomes, we performed a genome-wide integration site analysis using linker-mediated PCR (LM-PCR) and linear amplification-mediated PCR (LAM-PCR). Analysis of genomic DNA samples obtained from HSB5 treated female and male mice revealed a total of 1327 unique transposition events. Overall the chromosomal distribution pattern was close-to-random and we observed a random integration profile with respect to integration into gene and non-gene areas. Notably, when using the LM-PCR protocol, 27 extra-chromosomal integration events were identified, most likely caused by transposon excision and subsequent transposition into the delivered adenoviral vector genome. In total, this study provides a careful evaluation of the safety profile of adenovirus/Sleeping Beauty transposase hybrid-vectors. The obtained information will be useful when designing future preclinical studies utilizing hybrid-vectors in small and large animal models.
Keywords:	Adenoviridae, Computational Biology, Enzyme-Linked Immunosorbent Assay, Genetic Vectors, HeLa Cells, Transposases, Animals, Mice
Source:	PLoS ONE
ISSN:	1932-6203
Publisher:	Public Library of Science
Volume:	8
Number:	10
Page Range:	e75344
Date:	4 October 2013
Additional Information:	Erratum in: PLoS One. 2020 Feb 4;15(2):e0228902.
Official Publication:	https://doi.org/10.1371/journal.pone.0075344
PubMed:	View item in PubMed

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