Molecular Therapy: Methods & Clinical Development (Mar 2021)
CRISPR-Cas9 gene editing of hepatitis B virus in chronically infected humanized mice
- Daniel Stone,
- Kelly R. Long,
- Michelle A. Loprieno,
- Harshana S. De Silva Feelixge,
- Elizabeth J. Kenkel,
- R. Matt Liley,
- Stephen Rapp,
- Pavitra Roychoudhury,
- Thuy Nguyen,
- Laurence Stensland,
- Rossana Colón-Thillet,
- Lindsay M. Klouser,
- Nicholas D. Weber,
- Connie Le,
- Jessica Wagoner,
- Erin A. Goecker,
- Alvason Zhenhua Li,
- Karsten Eichholz,
- Lawrence Corey,
- D. Lorne Tyrrell,
- Alexander L. Greninger,
- Meei-Li Huang,
- Stephen J. Polyak,
- Martine Aubert,
- John E. Sagartz,
- Keith R. Jerome
Affiliations
- Daniel Stone
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Kelly R. Long
- Seventh Wave Laboratories, LLC, 19 Worthington Access Drive, Maryland Heights, MO 63043, USA
- Michelle A. Loprieno
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Harshana S. De Silva Feelixge
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Elizabeth J. Kenkel
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- R. Matt Liley
- Seventh Wave Laboratories, LLC, 19 Worthington Access Drive, Maryland Heights, MO 63043, USA
- Stephen Rapp
- Seventh Wave Laboratories, LLC, 19 Worthington Access Drive, Maryland Heights, MO 63043, USA
- Pavitra Roychoudhury
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Thuy Nguyen
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Laurence Stensland
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Rossana Colón-Thillet
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Lindsay M. Klouser
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Nicholas D. Weber
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Connie Le
- Li Ka Shing Institute of Virology, Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2E1, Canada
- Jessica Wagoner
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Erin A. Goecker
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Alvason Zhenhua Li
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Karsten Eichholz
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- Lawrence Corey
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- D. Lorne Tyrrell
- Li Ka Shing Institute of Virology, Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2E1, Canada
- Alexander L. Greninger
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Meei-Li Huang
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Stephen J. Polyak
- Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA
- Martine Aubert
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
- John E. Sagartz
- Seventh Wave Laboratories, LLC, 19 Worthington Access Drive, Maryland Heights, MO 63043, USA
- Keith R. Jerome
- Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA; Department of Laboratory Medicine, University of Washington, Seattle, WA 98195, USA; Corresponding author: Keith R. Jerome, MD, PhD, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue N, Seattle, WA 98109, USA.
- Journal volume & issue
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Vol. 20
pp. 258 – 275
Abstract
Chronic hepatitis B virus (HBV) infection is a major public health problem. New treatment approaches are needed because current treatments do not target covalently closed circular DNA (cccDNA), the template for HBV replication, and rarely clear the virus. We harnessed adeno-associated virus (AAV) vectors and CRISPR-Staphylococcus aureus (Sa)Cas9 to edit the HBV genome in liver-humanized FRG mice chronically infected with HBV and receiving entecavir. Gene editing was detected in livers of five of eight HBV-specific AAV-SaCas9-treated mice, but not control mice, and mice with detectable HBV gene editing showed higher levels of SaCas9 delivery to HBV+ human hepatocytes than those without gene editing. HBV-specific AAV-SaCas9 therapy significantly improved survival of human hepatocytes, showed a trend toward decreasing total liver HBV DNA and cccDNA, and was well tolerated. This work provides evidence for the feasibility and safety of in vivo gene editing for chronic HBV infections, and it suggests that with further optimization, this approach may offer a plausible way to treat or even cure chronic HBV infections.
