TY - JOUR
T1 - Molecular clones of genetically distinct hepatitis B virus genotypes reveal distinct host and drug treatment responses
AU - Liu, Yongzhen
AU - Park, Debby
AU - Cafiero, Thomas R.
AU - Bram, Yaron
AU - Chandar, Vasuretha
AU - Tseng, Anna
AU - Gertje, Hans P.
AU - Crossland, Nicholas A.
AU - Su, Lishan
AU - Schwartz, Robert E.
AU - Ploss, Alexander
N1 - Funding Information:
We thank Dr. Ju-Tao Guo (The Blumberg Institute) and Dr. Christoph Seeger (Fox Chase Cancer Center) for providing the rabbit anti-HBc polyclonal antibody and HepG2.2.15 cells, respectively. Dr. Peter Revill kindly provided a plasmid encoding the 1x HBV genotype B genome. We thank all members of the Ploss lab, in particular Drs. Lei Wei, Glenn Hogan, Emily Mesev and Robert LeDesma for critical discussion of the data and manuscript. This study was funded in part by grants from the National Institutes of Health R01AI138797 to L.S. and A.P.), ( R01AI107301 , R01AI146917 , R01AI153236 , R01AI168048 to A.P.) and ( R01DK121072 to R.E.S.), a Research Scholar Award from the American Cancer Society (RSG-15-048-01-MPC to A.P.), an Irma Hirschl Trust Research Scholar Award (R.E.S), a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (101539 A.P.), and funds from Princeton University . This work also utilized instruments acquired from NIH SIG grants ( S10OD026983 & S10OD030269 to N.A.C.).
Funding Information:
This study was funded in part by grants from the National Institutes of Health ( R01AI138797 to L.S. and A.P.), ( R01AI107301 , R01AI146917 , R01AI153236, R01AI168048 to A.P.) and ( R01DK121072 all to R.E.S.), a Research Scholar Award from the American Cancer Society (RSG-15-048-01-MPC to A.P.), a Irma Hirschl Trust Research Scholar Award (R.E.S), a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (101539 A.P.), and funds from Princeton University . This work also utilized an instrument acquired from a NIH SIG grants ( S10OD026983 & S10OD030269 to N.A.C.).
Funding Information:
This study was funded in part by grants from the National Institutes of Health (R01AI138797 to L.S. and A.P.), (R01AI107301, R01AI146917, R01AI153236, R01AI168048 to A.P.) and (R01DK121072 all to R.E.S.), a Research Scholar Award from the American Cancer Society (RSG-15-048-01-MPC to A.P.), a Irma Hirschl Trust Research Scholar Award (R.E.S), a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (101539 A.P.), and funds from Princeton University. This work also utilized an instrument acquired from a NIH SIG grants (S10OD026983 & S10OD030269 to N.A.C.).We thank Dr. Ju-Tao Guo (The Blumberg Institute) and Dr. Christoph Seeger (Fox Chase Cancer Center) for providing the rabbit anti-HBc polyclonal antibody and HepG2.2.15 cells, respectively. Dr. Peter Revill kindly provided a plasmid encoding the 1x HBV genotype B genome. We thank all members of the Ploss lab, in particular Drs. Lei Wei, Glenn Hogan, Emily Mesev and Robert LeDesma for critical discussion of the data and manuscript. This study was funded in part by grants from the National Institutes of Health R01AI138797 to L.S. and A.P.), (R01AI107301, R01AI146917, R01AI153236, R01AI168048 to A.P.) and (R01DK121072 to R.E.S.), a Research Scholar Award from the American Cancer Society (RSG-15-048-01-MPC to A.P.), an Irma Hirschl Trust Research Scholar Award (R.E.S), a Burroughs Wellcome Fund Award for Investigators in Pathogenesis (101539 A.P.), and funds from Princeton University. This work also utilized instruments acquired from NIH SIG grants (S10OD026983 & S10OD030269 to N.A.C.).
Publisher Copyright:
© 2022 The Authors
PY - 2022/9
Y1 - 2022/9
N2 - Background & Aims: HBV exhibits wide genetic diversity with at least 9 genotypes (GTs), which differ in terms of prevalence, geographic distribution, natural history, disease progression, and treatment outcome. However, differences in HBV replicative capacity, gene expression, and infective capability across different GTs remain incompletely understood. Herein, we aimed to study these crucial aspects using newly constructed infectious clones covering the major HBV GTs. Methods: The replicative capacity of infectious clones covering HBV GTs A-E was analyzed in cell lines, primary hepatocytes and humanized mice. Host responses and histopathology induced by the different HBV GTs were characterized in hydrodynamically injected mice. Differences in treatment responses to entecavir and various HBV capsid inhibitors were also quantified across the different genetically defined GTs. Results: Patient-derived HBV infectious clones replicated robustly both in vitro and in vivo. GTs A and D induce more pronounced intrahepatic and proinflammatory cytokine responses which correlated with faster viral clearance. Notably, all 5 HBV clones robustly produced viral particles following transfection into HepG2 cells, and these particles were infectious in HepG2-NTCP cells, primary human hepatocytes and human chimeric mice. Notably, GT D virus exhibited higher infectivity than GTs A, B, C and E in vitro, although it was comparable to GT A and B in the human liver chimeric mice in vivo. HBV capsid inhibitors were more readily capable of suppressing HBV GTs A, B, D and E than C. Conclusions: The infectious clones described here have broad utility as genetic tools that can mechanistically dissect intergenotypic differences in antiviral immunity and pathogenesis and aid in HBV drug development and screening. Lay summary: The hepatitis B virus (HBV) is a major contributor to human morbidity and mortality. HBV can be categorized into a number of genotypes, based on their specific genetic make-up, of which 9 are well known. We isolated and cloned the genomes of 5 of these genotypes and used them to create valuable tools for future research on this clinically important virus.
AB - Background & Aims: HBV exhibits wide genetic diversity with at least 9 genotypes (GTs), which differ in terms of prevalence, geographic distribution, natural history, disease progression, and treatment outcome. However, differences in HBV replicative capacity, gene expression, and infective capability across different GTs remain incompletely understood. Herein, we aimed to study these crucial aspects using newly constructed infectious clones covering the major HBV GTs. Methods: The replicative capacity of infectious clones covering HBV GTs A-E was analyzed in cell lines, primary hepatocytes and humanized mice. Host responses and histopathology induced by the different HBV GTs were characterized in hydrodynamically injected mice. Differences in treatment responses to entecavir and various HBV capsid inhibitors were also quantified across the different genetically defined GTs. Results: Patient-derived HBV infectious clones replicated robustly both in vitro and in vivo. GTs A and D induce more pronounced intrahepatic and proinflammatory cytokine responses which correlated with faster viral clearance. Notably, all 5 HBV clones robustly produced viral particles following transfection into HepG2 cells, and these particles were infectious in HepG2-NTCP cells, primary human hepatocytes and human chimeric mice. Notably, GT D virus exhibited higher infectivity than GTs A, B, C and E in vitro, although it was comparable to GT A and B in the human liver chimeric mice in vivo. HBV capsid inhibitors were more readily capable of suppressing HBV GTs A, B, D and E than C. Conclusions: The infectious clones described here have broad utility as genetic tools that can mechanistically dissect intergenotypic differences in antiviral immunity and pathogenesis and aid in HBV drug development and screening. Lay summary: The hepatitis B virus (HBV) is a major contributor to human morbidity and mortality. HBV can be categorized into a number of genotypes, based on their specific genetic make-up, of which 9 are well known. We isolated and cloned the genomes of 5 of these genotypes and used them to create valuable tools for future research on this clinically important virus.
KW - drug development
KW - genotypes
KW - hepatitis B
KW - hepatitis B virus
KW - host responses
KW - reverse genetics
KW - viral hepatitis
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UR - http://www.scopus.com/inward/citedby.url?scp=85135936382&partnerID=8YFLogxK
U2 - 10.1016/j.jhepr.2022.100535
DO - 10.1016/j.jhepr.2022.100535
M3 - Article
C2 - 36035359
AN - SCOPUS:85135936382
SN - 2589-5559
VL - 4
JO - JHEP Reports
JF - JHEP Reports
IS - 9
M1 - 100535
ER -