Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice

Kiourtis, C. , Wilczynska, A., Nixon, C., Clark, W., May, S. and Bird, T. G. (2021) Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice. Biology Open, 10(9), bio058678. (doi: 10.1242/bio.058678) (PMID:34435198) (PMCID:PMC8487635)

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Abstract

Mice are a widely used pre-clinical model system in large part due to their potential for genetic manipulation. The ability to manipulate gene expression in specific cells under temporal control is a powerful experimental tool. The liver is central to metabolic homeostasis and a site of many diseases, making the targeting of hepatocytes attractive. Adeno-Associated Virus 8 (AAV8) vectors are valuable instruments for the manipulation of hepatocellular gene expression. However, their off-target effects in mice have not been thoroughly explored. Here, we sought to identify the short-term off-target effects of AAV8 administration in mice. To do this, we injected C57BL/6J Wild-Type mice with either recombinant AAV8 vectors expressing Cre recombinase or control AAV8 vectors and characterised the changes in general health and in liver physiology, histology and transcriptomics compared to uninjected controls. We observed an acute and transient trend for reduction in homeostatic liver proliferation together with induction of the DNA damage marker γH2AX following AAV8 administration. The latter was enhanced upon Cre recombinase expression by the vector. Furthermore, we observed transcriptional changes in genes involved in circadian rhythm and response to infection. Notably, there were no additional transcriptomic changes upon expression of Cre recombinase by the AAV8 vector. Overall, there was no evidence of liver injury, and only mild T-cell infiltration was observed 28 days following AAV8 infection. These data advance the technique of hepatocellular genome editing through Cre-Lox recombination using Cre expressing AAV vectors, demonstrating their minimal effects on murine physiology and highlight the more subtle off target effects of these systems.

Item Type:Articles
Additional Information:CK, AW, CN, WC and SM were funded by Cancer Research UK (Grant number: A17196). TGB was funded by the Wellcome Trust (Grant number: WT107492Z).
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:May, Dr Steph and Nixon, Mr Colin and Bird, Dr Thomas and Clark, Mr William and Wilczynska, Dr Ania and Kiourtis, Christos
Authors: Kiourtis, C., Wilczynska, A., Nixon, C., Clark, W., May, S., and Bird, T. G.
College/School:College of Medical Veterinary and Life Sciences > School of Cancer Sciences
Journal Name:Biology Open
Publisher:Company of Biologists
ISSN:2046-6390
ISSN (Online):2046-6390
Published Online:26 August 2021
Copyright Holders:Copyright © 2021 The Company of Biologists Ltd
First Published:First published in Biology Open 10(9): bio058678
Publisher Policy:Reproduced under a Creative Commons License

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