Liver-specific transgenic expression of human NTCP in rhesus macaques confers HBV susceptibility on primary hepatocytes

L Lauren N. Rust (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) J Jochen M. Wettengel (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) S Sreya Biswas (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) J Junghyun Ryu (Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University) N Nadine Piekarski (Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University) S Sofiya Yusova (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) S Savannah S. Lutz (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) S Spandana Naldiga (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) B Brayden J. Hinrichs (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) M Michelle N. Sullivan (Division of Comparative Medicine, Oregon National Primate Research Center, Oregon Health and Science University) J Jamie O. Lo (Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University) U Ulrike Protzer (Institute of Virology, Technical University of Munich/Helmholtz Munich) J Jeremy V. Smedley (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) J Jonah B. Sacha (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University) C Carol B. Hanna (Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University) B Benjamin N. Bimber (Division of Genetics, Oregon National Primate Research Center, Oregon Health and Science University) J Jon D. Hennebold (Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center) B Benjamin J. Burwitz (Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University)

Abstract

Hepatitis B virus (HBV) poses a significant global health challenge, necessitating the urgent development of curative therapeutics. However, this progress is impeded by the lack of robust, immunocompetent preclinical animal models due to HBV’s strict species specificity. We previously showed that vector-mediated expression of the HBV entry receptor, human sodium-taurocholate cotransporting polypeptide (hNTCP), renders macaques fully susceptible to HBV infection. In this study, we have generated transgenic macaques expressing hNTCP, marking the creation of the first transgenic nonhuman primate model for infectious disease research. We used PiggyBac (PB) transposon technology to insert a liver-specific hNTCP expression cassette into rhesus macaque zygotes and transferred the resulting embryos into surrogate females, resulting in two healthy transgenic offspring. In both animals, hNTCP is highly and selectively expressed in the liver. Most importantly, we show that isolated hepatocytes from these monkeys are susceptible to HBV infection. These findings lay the foundation for the development of a nonhuman primate HBV model, facilitating the advancement and validation of curative HBV therapies.

Article Details

Volume / Issue Vol. 122, Issue 7
Published February 18, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (18)

L

Lauren N. Rust

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

J

Jochen M. Wettengel

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

S

Sreya Biswas

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

J

Junghyun Ryu

Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University

N

Nadine Piekarski

Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University

S

Sofiya Yusova

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

S

Savannah S. Lutz

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

S

Spandana Naldiga

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

B

Brayden J. Hinrichs

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

M

Michelle N. Sullivan

Division of Comparative Medicine, Oregon National Primate Research Center, Oregon Health and Science University

J

Jamie O. Lo

Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University

U

Ulrike Protzer

Institute of Virology, Technical University of Munich/Helmholtz Munich

J

Jeremy V. Smedley

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

J

Jonah B. Sacha

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University

C

Carol B. Hanna

Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health and Science University

B

Benjamin N. Bimber

Division of Genetics, Oregon National Primate Research Center, Oregon Health and Science University

J

Jon D. Hennebold

Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center

B

Benjamin J. Burwitz

Division of Pathobiology and Immunology, Oregon National Primate Research Center, Oregon Health and Science University