Advertisement

AAV-Mediated Gene Delivery to the Mouse Liver

  • Sharon C. Cunningham
  • Ian E. AlexanderEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1937)

Abstract

The liver is an attractive target for gene therapy due to the high incidence of liver disease phenotypes. Adeno-associated viral vectors (AAV) are currently the most popular gene delivery system for targeting the liver, reflecting high transduction efficiency in vivo and the availability of a toolkit of multiple different capsids with high liver tropism. While AAV vectors confer stable gene transfer in the relatively quiescent adult liver, the predominantly episomal nature of AAV vector genomes results in less stable expression in the growing liver as a consequence of episome clearance during hepatocellular replication. This is an important consideration in experimental design involving young animals, particularly mice, where liver growth is rapid. Given the immense value of murine models for dissecting disease pathophysiology, experimental therapeutics and vector development, this technical manuscript focuses on AAV-mediated transduction of the mouse liver. Xenograft models, in which chimeric mouse-human livers can be established, are also amenable to AAV-mediated gene transfer and have proven to be powerful tools for in vivo selection and characterization of novel human-specific capsids. While yet to be confirmed, such models have the potential to more accurately predict transduction efficiency of clinical candidate vectors than nonhuman primate models.

Key words

Mouse Liver Hepatocyte Gene transfer Adeno-associated virus Viral vector Metabolic Hepatocellular Transposon 

Notes

Acknowledgments

This work was supported by NHMRC grants APP1008021 and APP1065053 to I.E.A.

References

  1. 1.
    Alexander IE, Kok C, Dane AP et al (2012) Gene therapy for metabolic disorders: an overview with a focus on urea cycle disorders. J Inherit Metab Dis 35(4):641–645CrossRefGoogle Scholar
  2. 2.
    Laurence JM, Allen RD, McCaughan GW et al (2009) Gene therapy in transplantation. Transplant Rev (Orlando) 23(3):159–170CrossRefGoogle Scholar
  3. 3.
    Gao GP, Alvira MR, Wang L et al (2002) Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc Natl Acad Sci U S A 99:11854–11859CrossRefGoogle Scholar
  4. 4.
    Davidoff AM, Gray JT, Ng CY et al (2005) Comparison of the ability of adeno-associated viral vectors pseudotyped with serotype 2, 5, and 8 capsid proteins to mediate efficient transduction of the liver in murine and nonhuman primate models. Mol Ther 11:875–888CrossRefGoogle Scholar
  5. 5.
    Nathwani AC, Gray JT, Ng CY et al (2006) Self-complementary adeno-associated virus vectors containing a novel liver-specific human factor IX expression cassette enable highly efficient transduction of murine and nonhuman primate liver. Blood 107:2653–2661CrossRefGoogle Scholar
  6. 6.
    Nathwani AC, Reiss UM, Tuddenham EG et al (2014) Long-term safety and efficacy of factor IX gene therapy in haemophilia B. N Engl J Med 371(21):1994–2004CrossRefGoogle Scholar
  7. 7.
    Lisowski L, Dane AP, Chu K et al (2014) Selection and evaluation of clinically relevant AAV variants in a xenograft liver model. Nature 506(7488):382–386CrossRefGoogle Scholar
  8. 8.
    Azuma H, Paulk N, Ranade A et al (2007) Robust expansion of human hepatocytes in Fah−/−/Rag2−/−/Il2rg−/− mice. Nat Biotechnol 25:903–910CrossRefGoogle Scholar
  9. 9.
    Paulk NK, Pekrun K, Zhu E et al (2018) Bioengineered AAV capsids with combined high human liver transduction in vivo and unique humoral seroreactivity. Mol Ther 26(1):289–303CrossRefGoogle Scholar
  10. 10.
    Nakai H, Montini E, Fuess S et al (2003) AAV serotype 2 vectors preferentially integrate into active genes in mice. Nat Genet 34(2):297–302CrossRefGoogle Scholar
  11. 11.
    Cunningham SC, Dane AP, Spinoulas A et al (2008) Gene delivery to the juvenile mouse liver using AAV2/8 vectors. Mol Ther 16(6):1081–1088CrossRefGoogle Scholar
  12. 12.
    Nakai H, Yant SR, Storm TA et al (2001) Extrachromosomal recombinant adeno-associated virus vector genomes are primarily responsible for stable liver transduction in vivo. J Virol 75:6969–6976CrossRefGoogle Scholar
  13. 13.
    Wang Z, Zhu T, Qiao C et al (2005) Adeno-associated virus serotype 8 efficiency delivers genes to muscle and heart. Nat Biotechnol 23:321–328CrossRefGoogle Scholar
  14. 14.
    Cunningham SC, Spinoulas A, Carpenter KH et al (2009) AAV2/8-mediated correction of OTC deficiency is robust in adult but not neonatal spfash mice. Mol Ther 17(8):1340–1346CrossRefGoogle Scholar
  15. 15.
    Cunningham SC, Kok CY, Dane AP et al (2011) Induction and prevention of severe hyperammonemia in the spfash mouse model of ornithine transcarbamylase deficiency using shRNA and rAAV-mediated gene delivery. Mol Ther 19(5):854–859CrossRefGoogle Scholar
  16. 16.
    Cunningham SC, Kok CY, Spinoulas A et al (2013) AAV-encoded OTC activity persisting to adulthood following delivery to newborn spfash mice is insufficient to prevent shRNA-induced hyperammonaemia. Gene Ther 20(12):1184–1187CrossRefGoogle Scholar
  17. 17.
    Kok CY, Cunningham SC, Carpenter KH et al (2013) Adeno-associated virus-mediated rescue of neonatal lethality in argininosuccinate synthetase-deficient mice. Mol Ther 21(10):1823–1831CrossRefGoogle Scholar
  18. 18.
    Cunningham SC, Siew SM, Hallwirth CV et al (2015) Modeling correction of severe urea cycle defects in the growing murine liver using a hybrid recombinant adeno-associated virus/piggyBac transposase gene delivery system. Hepatology 62:417–428CrossRefGoogle Scholar
  19. 19.
    Lukas G, Brindle SD, Greengard P (1971) The route of absorption of intraperitoneally administered compounds. J Pharmacol Exp Ther 178(3):562–564PubMedGoogle Scholar
  20. 20.
    Dane AP, Wowro SJ, Cunningham SC et al (2013) Comparison of gene transfer to the murine liver following intraperitoneal and intraportal delivery of hepatotropic AAV pseudo-serotypes. Gene Ther 20(4):460–464CrossRefGoogle Scholar
  21. 21.
    Vollmar B, Menger MD (2009) The hepatic microcirculation: mechanistic contributions and therapeutic targets in liver injury and repair. Physiol Rev 89:1269–1339CrossRefGoogle Scholar
  22. 22.
    Graham FL, Smiley J, Russell WC et al (1977) Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol 36:59–74CrossRefGoogle Scholar
  23. 23.
    Grieger JC, Choi VW, Samulski RJ (2006) Production and characterization of adeno-associated viral vectors. Nat Protoc 1:1412–1428CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Gene Therapy Research Unit, Children’s Medical Research InstituteThe University of Sydney, Faculty of Medicine and Health and Sydney Children’s Hospitals NetworkWestmeadAustralia
  2. 2.The University of Sydney, Sydney Medical SchoolDiscipline of Child and Adolescent HealthWestmeadAustralia

Personalised recommendations