AAV capsid prioritization in normal and steatotic human livers maintained by machine perfusion
Therapeutic efficacy and safety of adeno-associated virus (AAV) liver gene therapy depend on capsid choice. To predict AAV capsid performance under near-clinical conditions, we established side-by-side comparison at single-cell resolution in human livers maintained by normothermic machine perfusion. AAV-LK03 transduced hepatocytes much more efficiently and specifically than AAV5, AAV8 and AAV6, which are most commonly used clinically, and AAV-NP59, which is better at transducing human hepatocytes engrafted in immune-deficient mice. AAV-LK03 preferentially transduced periportal hepatocytes in normal liver, whereas AAV5 targeted pericentral hepatocytes in steatotic liver. AAV5 and AAV8 transduced liver sinusoidal endothelial cells as efficiently as hepatocytes. AAV capsid and steatosis influenced vector episome formation, which determines gene therapy durability, with AAV5 delaying concatemerization. Our findings inform capsid choice in clinical AAV liver gene therapy, including consideration of disease-relevant hepatocyte zonation and effects of steatosis, and facilitate the development of AAV capsids that transduce hepatocytes or other therapeutically relevant cell types in the human liver with maximum efficiency and specificity.
Authors
- Miguel Nunez
- Holger Willenbring (ORCID: https://orcid.org/0000-0002-7397-2957)
- Julia Driscoll (ORCID: https://orcid.org/0000-0003-4535-8463)
- Sandeep K. Dhillon
- Shareef Syed (ORCID: https://orcid.org/0000-0002-1475-778X)
- Amita Tiyaboonchai (ORCID: https://orcid.org/0000-0002-0116-7393)
- Jacquelyn J. Maher (ORCID: https://orcid.org/0000-0002-2509-8484)
- Abhishek Murti (ORCID: https://orcid.org/0000-0003-0198-9933)
- Aras N. Mattis (ORCID: https://orcid.org/0000-0002-5813-164X)
- Tyler Lunow-Luke (ORCID: https://orcid.org/0000-0001-5920-4313)
- Simone N. T. Kurial (ORCID: https://orcid.org/0000-0003-1175-6076)
- Jan Bartel (ORCID: https://orcid.org/0000-0001-7602-7224)
- Markus Grompe (ORCID: https://orcid.org/0000-0002-6616-4345)
- Pervinder K. Choksi
- W. Yue (ORCID: https://orcid.org/0000-0003-2601-9378)
- Chris Her
- Garrett R. Roll (ORCID: https://orcid.org/0000-0001-6241-7037)
- Tin K. Mao
- Bruce M. Wang
- Jae-Jun Kim (ORCID: https://orcid.org/0000-0003-3222-5506)
- Julian N. Ramos
Institutions
- Oregon Health & Science University (US)
- University of California, San Francisco (US)
- University of California, Irvine (US)
- Pliant (United States) (US)
- Broad Center (US)
Publication Details
- Journal
- Nature Biotechnology
- Published
- 2025-01-29
- DOI
- https://doi.org/10.1038/s41587-024-02523-6
- Citations
- 24
- Primary Topic
- Virus-based gene therapy research
- Type
- article
- Field-Weighted Citation Impact
- 43.61
Funders
- California Institute for Regenerative Medicine
- University of California, San Francisco
- National Institute of Diabetes and Digestive and Kidney Diseases