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

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 disease...

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Autores principales:	Christos Kiourtis, Ania Wilczynska, Colin Nixon, William Clark, Stephanie May, Thomas G. Bird
Formato:	article
Lenguaje:	EN
Publicado:	The Company of Biologists 2021
Materias:	adeno-associated virus liver disease mouse model genetic models Science Q Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/20782b67b1f6464d846e370308cf1da4
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spelling	oai:doaj.org-article:20782b67b1f6464d846e370308cf1da42021-11-28T16:01:11ZSpecificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice2046-639010.1242/bio.058678https://doaj.org/article/20782b67b1f6464d846e370308cf1da42021-09-01T00:00:00Zhttp://bio.biologists.org/content/10/9/bio058678https://doaj.org/toc/2046-6390Mice 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 14 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.Christos KiourtisAnia WilczynskaColin NixonWilliam ClarkStephanie MayThomas G. BirdThe Company of Biologistsarticleadeno-associated virusliver diseasemouse modelgenetic modelsScienceQBiology (General)QH301-705.5ENBiology Open, Vol 10, Iss 9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	adeno-associated virus liver disease mouse model genetic models Science Q Biology (General) QH301-705.5
spellingShingle	adeno-associated virus liver disease mouse model genetic models Science Q Biology (General) QH301-705.5 Christos Kiourtis Ania Wilczynska Colin Nixon William Clark Stephanie May Thomas G. Bird Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice
description	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 14 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.
format	article
author	Christos Kiourtis Ania Wilczynska Colin Nixon William Clark Stephanie May Thomas G. Bird
author_facet	Christos Kiourtis Ania Wilczynska Colin Nixon William Clark Stephanie May Thomas G. Bird
author_sort	Christos Kiourtis
title	Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice
title_short	Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice
title_full	Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice
title_fullStr	Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice
title_full_unstemmed	Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice
title_sort	specificity and off-target effects of aav8-tbg viral vectors for the manipulation of hepatocellular gene expression in mice
publisher	The Company of Biologists
publishDate	2021
url	https://doaj.org/article/20782b67b1f6464d846e370308cf1da4
work_keys_str_mv	AT christoskiourtis specificityandofftargeteffectsofaav8tbgviralvectorsforthemanipulationofhepatocellulargeneexpressioninmice AT aniawilczynska specificityandofftargeteffectsofaav8tbgviralvectorsforthemanipulationofhepatocellulargeneexpressioninmice AT colinnixon specificityandofftargeteffectsofaav8tbgviralvectorsforthemanipulationofhepatocellulargeneexpressioninmice AT williamclark specificityandofftargeteffectsofaav8tbgviralvectorsforthemanipulationofhepatocellulargeneexpressioninmice AT stephaniemay specificityandofftargeteffectsofaav8tbgviralvectorsforthemanipulationofhepatocellulargeneexpressioninmice AT thomasgbird specificityandofftargeteffectsofaav8tbgviralvectorsforthemanipulationofhepatocellulargeneexpressioninmice
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Specificity and off-target effects of AAV8-TBG viral vectors for the manipulation of hepatocellular gene expression in mice

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