@article{745e3458187a4641ac708f6b167907b0,
title = "AAV integration in human hepatocytes",
abstract = "Recombinant adeno-associated viral (rAAV) vectors are considered promising tools for gene therapy directed at the liver. Whereas rAAV is thought to be an episomal vector, its single-stranded DNA genome is prone to intra- and inter-molecular recombination leading to rearrangements and integration into the host cell genome. Here, we ascertained the integration frequency of rAAV in human hepatocytes transduced either ex vivo or in vivo and subsequently expanded in a mouse model of xenogeneic liver regeneration. Chromosomal rAAV integration events and vector integrity were determined using the capture-PacBio sequencing approach, a long-read next-generation sequencing method that has not previously been used for this purpose. Chromosomal integrations were found at a surprisingly high frequency of 1%–3% both in vitro and in vivo. Importantly, most of the inserted rAAV sequences were heavily rearranged and were accompanied by deletions of the host genomic sequence at the integration site.",
keywords = "FRGN, capture sequencing, genotoxicity, rAAV, random integration",
author = "Dalwadi, {Dhwanil A.} and Andrea Calabria and Amita Tiyaboonchai and Jeffrey Posey and Naugler, {Willscott E.} and Eugenio Montini and Markus Grompe",
note = "Funding Information: We thank Carlo Cipriani for the help on bioinformatics revisions. This research was funded by the National Institutes of Health grant R01CA190144 to W.E.N. and M.G. D.A.D. and A.T. designed and performed experiments, analyzed data, and helped write the manuscript. A.C. designed and performed bioinformatics analysis and helped write the manuscript. J.P. produced and quantified AAV. M.G. and W.E.N. conceived the project, helped design experiments, and helped write the manuscript. E.M. analyzed bioinformatics and helped write the manuscript. M.G. has a financial interest in Yecuris Corp. (Tigard, Oregon), a company that has commercialized the humanized mouse liver model used in this work. MG also is a consultant for LogicBio Therapeutics. Other authors declare no competing interests. Funding Information: We thank Carlo Cipriani for the help on bioinformatics revisions. This research was funded by the National Institutes of Health grant R01CA190144 to W.E.N. and M.G. Publisher Copyright: {\textcopyright} 2021 The American Society of Gene and Cell Therapy",
year = "2021",
month = oct,
day = "6",
doi = "10.1016/j.ymthe.2021.08.031",
language = "English (US)",
volume = "29",
pages = "2898--2909",
journal = "Molecular Therapy",
issn = "1525-0016",
publisher = "Nature Publishing Group",
number = "10",
}