Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice
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 transduct...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-09-01
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| Series: | Molecular Therapy: Methods & Clinical Development |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2329050120301200 |
| _version_ | 1818154679983407104 |
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| author | Chenhui Zou Koen O.A. Vercauteren Eleftherios Michailidis Mohammad Kabbani Irene Zoluthkin Corrine Quirk Luis Chiriboga Mustafa Yazicioglu Xavier M. Anguela Philip Meuleman Katherine A. High Roland W. Herzog Ype P. de Jong |
| author_facet | Chenhui Zou Koen O.A. Vercauteren Eleftherios Michailidis Mohammad Kabbani Irene Zoluthkin Corrine Quirk Luis Chiriboga Mustafa Yazicioglu Xavier M. Anguela Philip Meuleman Katherine A. High Roland W. Herzog Ype P. de Jong |
| author_sort | Chenhui Zou |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-11T14:30:21Z |
| format | Article |
| id | doaj.art-76e8a94fb75547cf982881d7ea650537 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-12-11T14:30:21Z |
| publishDate | 2020-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-76e8a94fb75547cf982881d7ea6505372022-12-22T01:02:27ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012020-09-0118189198Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric MiceChenhui Zou0Koen O.A. Vercauteren1Eleftherios Michailidis2Mohammad Kabbani3Irene Zoluthkin4Corrine Quirk5Luis Chiriboga6Mustafa Yazicioglu7Xavier M. Anguela8Philip Meuleman9Katherine A. High10Roland W. Herzog11Ype P. de Jong12Division of Gastroenterology and Hepatology, Weill Cornell Medicine, New York, NY 10065, USA; Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY 10065, USALaboratory of Virology and Infectious Disease, Rockefeller University, New York, NY 10065, USA; Laboratory of Liver Infectious Diseases, Ghent University, 9000 Ghent, BelgiumLaboratory of Virology and Infectious Disease, Rockefeller University, New York, NY 10065, USALaboratory of Virology and Infectious Disease, Rockefeller University, New York, NY 10065, USADepartment of Pediatrics, University of Florida College of Medicine, Gainesville, FL 32603, USALaboratory of Virology and Infectious Disease, Rockefeller University, New York, NY 10065, USADepartment of Pathology, NYU Langone Health, New York, NY 10016, USASpark Therapeutics, Philadelphia, PA 19104, USASpark Therapeutics, Philadelphia, PA 19104, USALaboratory of Liver Infectious Diseases, Ghent University, 9000 Ghent, BelgiumSpark Therapeutics, Philadelphia, PA 19104, USADepartment of Pediatrics, Indiana University, Indianapolis, IN 46202, USA; Herman B Wells Center for Pediatric Research, IUPUI, Indianapolis, IN 46202, USADivision of Gastroenterology and Hepatology, Weill Cornell Medicine, New York, NY 10065, USA; Laboratory of Virology and Infectious Disease, Rockefeller University, New York, NY 10065, USA; Corresponding author: Ype P. de Jong, Division of Gastroenterology and Hepatology, Weill Cornell Medicine, 1305 York Ave., New York, NY 10065, USA.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.http://www.sciencedirect.com/science/article/pii/S2329050120301200humanized miceAAVgene therapyliverhuman hepatocytesdonor variability |
| spellingShingle | Chenhui Zou Koen O.A. Vercauteren Eleftherios Michailidis Mohammad Kabbani Irene Zoluthkin Corrine Quirk Luis Chiriboga Mustafa Yazicioglu Xavier M. Anguela Philip Meuleman Katherine A. High Roland W. Herzog Ype P. de Jong Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice Molecular Therapy: Methods & Clinical Development humanized mice AAV gene therapy liver human hepatocytes donor variability |
| title | Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice |
| title_full | Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice |
| title_fullStr | Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice |
| title_full_unstemmed | Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice |
| title_short | Experimental Variables that Affect Human Hepatocyte AAV Transduction in Liver Chimeric Mice |
| title_sort | experimental variables that affect human hepatocyte aav transduction in liver chimeric mice |
| topic | humanized mice AAV gene therapy liver human hepatocytes donor variability |
| url | http://www.sciencedirect.com/science/article/pii/S2329050120301200 |
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