Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice

• Published in: Molecular therapy. Methods & clinical development 2020-09, Vol.18, p.189-198
• Main Authors: Zou, Chenhui, Vercauteren, Koen O.A., Michailidis, Eleftherios, Kabbani, Mohammad, Zoluthkin, Irene, Quirk, Corrine, Chiriboga, Luis, Yazicioglu, Mustafa, Anguela, Xavier M., Meuleman, Philip, High, Katherine A., Herzog, Roland W., de Jong, Ype P.
• Format: Article
• Language: English
• Subjects: AAV; donor variability; gene therapy; hemophilia; human hepatocytes; humanized mice; liver
• ISSN: 2329-0501; 2329-0501
• DOI: 10.1016/j.omtm.2020.05.033

Adeno-associated virus (AAV) vector serotypes vary in their ability to transduce hepatocytes from different species. Chimeric mouse models harboring human hepatocytes have shown translational promise for liver-directed gene therapies. However, many variables that influence human hepatocyte transduction and transgene expression in such models remain poorly defined. Here, we aimed to test whether three experimental conditions influence AAV transgene expression in immunodeficient, fumaryl-acetoactetate-hydrolase-deficient (Fah−/−) chimeric mice repopulated with primary human hepatocytes. We examined the effects of the murine liver injury cycle, human donor variability, and vector doses on hepatocyte transduction with various AAV serotypes expressing a green fluorescent protein (GFP). We determined that the timing of AAV vector challenge in the liver injury cycle resulted in up to 7-fold differences in the percentage of GFP expressing human hepatocytes. The GFP+ hepatocyte frequency varied 7-fold between human donors without, however, changing the relative transduction efficiency between serotypes for an individual donor. There was also a clear relationship between AAV vector doses and human hepatocyte transduction and transgene expression. We conclude that several experimental variables substantially affect human hepatocyte transduction in the Fah−/− chimera model, attention to which may improve reproducibility between findings from different laboratories. [Display omitted] Liver chimeric mice are widely used to model human hepatocyte susceptibility to adeno-associated virus (AAV) gene vector transduction with inconsistent finding between laboratories. Zou et al. quantify to what extent three experimental conditions—namely, mouse liver injury, human donor variability, and AAV doses—affect human hepatocyte transduction in chimeric mice.