Human Hepatocyte Transduction with Adeno-Associated Virus Vector

Zhenwei Song; Wenwei Shao; Liujiang Song; Xieolei Pei; Chengwen Li

doi:10.1007/978-1-0716-2557-6_5

Human Hepatocyte Transduction with Adeno-Associated Virus Vector

Methods Mol Biol. 2022:2544:83-93. doi: 10.1007/978-1-0716-2557-6_5.

Authors

Zhenwei Song¹, Wenwei Shao², Liujiang Song¹, Xieolei Pei¹, Chengwen Li³

Affiliations

¹ Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
² Academy of Medical Engineering and Translational Medicine, Tianjin University, Tianjin, China.
³ Gene Therapy Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. chengwen_li@med.unc.edu.

PMID: 36125711
DOI: 10.1007/978-1-0716-2557-6_5

Abstract

As the adeno-associated virus (AAV) vectors hold unique advantages over other viral vectors, AAV gene therapy has accumulated rapid progress and development. Liver-targeted gene therapy by AAV vectors has been successfully applied in clinical trials for many diseases. Low transduction efficiency and high prevalence of neutralizing antibodies (Nabs), however, are the major obstacles to further translate this therapeutic strategy into clinical trials. Pre-clinical evaluation on hepatocytes could help to elucidate the tropism of AAV serotypes for liver-targeted gene therapy, and could also provide a test model to develop novel AAV mutants with Nabs evasion and high liver tropism. Here, we described the basic laboratory procedure to apply the AAV vector to transduce human hepatocytes in vitro and in vivo with some tips gained from our own experience.

Keywords: Adeno-associated virus vector (AAV); Liver chimeric mice; Primary human hepatocyte (PHH); Transduction.

MeSH terms

Antibodies, Neutralizing
Dependovirus* / genetics
Genetic Therapy / methods
Genetic Vectors* / genetics
Hepatocytes
Humans

Substances

Antibodies, Neutralizing