Binding of hepatitis B virus to its cellular receptor alters the expression profile of genes of bile acid metabolism

Chronic hepatitis B virus (HBV) infection has been associated with alterations in lipid metabolism. Moreover, the Na+‐taurocholate cotransporting polypeptide (NTCP), responsible for bile acid (BA) uptake into hepatocytes, was identified as the functional cellular receptor mediating HBV entry. The aim of the study was to determine whether HBV alters the liver metabolic profile by employing HBV‐infected and uninfected human liver chimeric mice. Humanized urokinase plasminogen activator/severe combined immunodeficiency mice were used to establish chronic HBV infection. Gene expression profiles were determined by real‐time polymerase chain reaction using primers specifically recognizing transcripts of either human or murine origin. Liver biopsy samples obtained from HBV‐chronic individuals were used to validate changes determined in mice. Besides modest changes in lipid metabolism, HBV‐infected mice displayed a significant enhancement of human cholesterol 7α‐hydroxylase (human [h]CYP7A1; median 12‐fold induction; P < 0.0001), the rate‐limiting enzyme promoting the conversion of cholesterol to BAs, and of genes involved in transcriptional regulation, biosynthesis, and uptake of cholesterol (human sterol‐regulatory element‐binding protein 2, human 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase, and human low‐density lipoprotein receptor), compared to uninfected controls. Significant hCYP7A1 induction and reduction of human small heterodimer partner, the corepressor of hCYP7A1 transcription, was also confirmed in liver biopsies from HBV‐infected patients. Notably, administration of Myrcludex‐B, an entry inhibitor derived from the pre‐S1 domain of the HBV envelope, provoked a comparable murine CYP7A1 induction in uninfected mice, thus designating the pre‐S1 domain as the viral component triggering such metabolic alterations. Conclusion: Binding of HBV to NTCP limits its function, thus promoting compensatory BA synthesis and cholesterol provision. The intimate link determined between HBV and liver metabolism underlines the importance to exploit further metabolic pathways, as well as possible NTCP‐related viral‐drug interactions. (Hepatology 2014;60:1483–1493)

[1]  U. Beuers,et al.  Impaired uptake of conjugated bile acids and hepatitis b virus pres1-binding in na+-taurocholate cotransporting polypeptide knockout mice , 2015, Hepatology.

[2]  H. Chan,et al.  Hepatitis B virus infection , 2014, The Lancet.

[3]  A. Geier.  Hepatitis B virus: The “metabolovirus” highjacks cholesterol and bile acid metabolism , 2014, Hepatology.

[4]  A. Geipel,et al.  Kinetics of the bile acid transporter and hepatitis B virus receptor Na+/taurocholate cotransporting polypeptide (NTCP) in hepatocytes. , 2014, Journal of hepatology.

[5]  P. Parini,et al.  Erratum: Mice with chimeric livers are an improved model for human lipoprotein metabolism (PLoS ONE (2013) 8, 11 (e78550) DOI: 10.1371/journal.pone.0078550) , 2014 .

[6]  R. Bartenschlager,et al.  Strategies to inhibit entry of HBV and HDV into hepatocytes. , 2014, Gastroenterology.

[7]  G. Patman.  Hepatitis: HBV infection alters bile acid metabolism gene profile , 2014, Nature Reviews Gastroenterology &Hepatology.

[8]  V. Lohmann,et al.  Cyclosporin A inhibits hepatitis B and hepatitis D virus entry by cyclophilin-independent interference with the NTCP receptor. , 2014, Journal of hepatology.

[9]  H. Kusuhara,et al.  Cyclosporin A and its analogs inhibit hepatitis B virus entry into cultured hepatocytes through targeting a membrane transporter, sodium taurocholate cotransporting polypeptide (NTCP) , 2014, Hepatology.

[10]  M. Fälth,et al.  Hepatitis B and D viruses exploit sodium taurocholate co-transporting polypeptide for species-specific entry into hepatocytes. , 2014, Gastroenterology.

[11]  S. Strom,et al.  Mice with Chimeric Livers Are an Improved Model for Human Lipoprotein Metabolism , 2013, PloS one.

[12]  T. Weiss,et al.  Myristoylated PreS1‐domain of the hepatitis B virus L‐protein mediates specific binding to differentiated hepatocytes , 2013, Hepatology.

[13]  U. Haberkorn,et al.  Hepatitis B virus hepatotropism is mediated by specific receptor recognition in the liver and not restricted to susceptible hosts , 2013, Hepatology.

[14]  Wenhui Li,et al.  Molecular Determinants of Hepatitis B and D Virus Entry Restriction in Mouse Sodium Taurocholate Cotransporting Polypeptide , 2013, Journal of Virology.

[15]  J. M. Suh,et al.  PPARγ signaling and metabolism: the good, the bad and the future , 2013, Nature Medicine.

[16]  A. Lohse,et al.  The entry inhibitor Myrcludex-B efficiently blocks intrahepatic virus spreading in humanized mice previously infected with hepatitis B virus. , 2013, Journal of hepatology.

[17]  Sean Ekins,et al.  Structure-activity relationship for FDA approved drugs as inhibitors of the human sodium taurocholate cotransporting polypeptide (NTCP). , 2013, Molecular pharmaceutics.

[18]  Wenhui Li,et al.  Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus , 2012, eLife.

[19]  M. Dandri,et al.  New insight in the pathobiology of hepatitis B virus infection , 2012, Gut.

[20]  Michael Thomaschewski,et al.  RGB marking with lentiviral vectors for multicolor clonal cell tracking , 2012, Nature Protocols.

[21]  A. Lohse,et al.  Humanized chimeric uPA mouse model for the study of hepatitis B and D virus interactions and preclinical drug evaluation , 2012, Hepatology.

[22]  T. Weiss,et al.  Hepatocyte polarization is essential for the productive entry of the hepatitis B virus , 2012, Hepatology.

[23]  T. Tseng,et al.  Impact of hepatitis B virus infection on metabolic profiles and modifying factors , 2012, Journal of viral hepatitis.

[24]  M. Dandri,et al.  Chimeric mouse model of hepatitis B virus infection. , 2012, Journal of hepatology.

[25]  R. Dwek,et al.  Cholesterol Depletion of Hepatoma Cells Impairs Hepatitis B Virus Envelopment by Altering the Topology of the Large Envelope Protein , 2011, Journal of Virology.

[26]  A. Lohse,et al.  Hepatitis B virus limits response of human hepatocytes to interferon-α in chimeric mice. , 2011, Gastroenterology.

[27]  Hyun Kook Cho,et al.  Oxygenated derivatives of cholesterol promote hepatitis B virus gene expression through nuclear receptor LXRα activation. , 2011, Virus research.

[28]  Y. Shaul,et al.  Hepatocyte metabolic signalling pathways and regulation of hepatitis B virus expression , 2011, Liver international : official journal of the International Association for the Study of the Liver.

[29]  M. Trauner,et al.  Nuclear receptors in liver disease , 2011, Hepatology.

[30]  J. Chiang,et al.  Overexpression of cholesterol 7α‐hydroxylase promotes hepatic bile acid synthesis and secretion and maintains cholesterol homeostasis , 2011, Hepatology.

[31]  Hyun Kook Cho,et al.  Bile acids increase hepatitis B virus gene expression and inhibit interferon‐α activity , 2010, The FEBS journal.

[32]  B. Fehse,et al.  Lentiviral gene ontology (LeGO) vectors equipped with novel drug-selectable fluorescent proteins: new building blocks for cell marking and multi-gene analysis , 2010, Gene Therapy.

[33]  F. Chisari,et al.  Human liver chimeric mice provide a model for hepatitis B and C virus infection and treatment. , 2010, The Journal of clinical investigation.

[34]  Y. Ni,et al.  Fine Mapping of Pre-S Sequence Requirements for Hepatitis B Virus Large Envelope Protein-Mediated Receptor Interaction , 2009, Journal of Virology.

[35]  M. Levrero,et al.  Control of cccDNA function in hepatitis B virus infection. , 2009, Journal of hepatology.

[36]  Y. Shin,et al.  Liver X receptor mediates hepatitis B virus X protein–induced lipogenesis in hepatitis B virus–associated hepatocellular carcinoma , 2009, Hepatology.

[37]  M. Hardt,et al.  Hepatitis B virus infection is dependent on cholesterol in the viral envelope , 2009, Cellular microbiology.

[38]  A. Tall,et al.  HDL, ABC transporters, and cholesterol efflux: implications for the treatment of atherosclerosis. , 2008, Cell metabolism.

[39]  U. Haberkorn,et al.  Prevention of hepatitis B virus infection in vivo by entry inhibitors derived from the large envelope protein , 2008, Nature Biotechnology.

[40]  A. Quaas,et al.  Impaired intrahepatic hepatitis B virus productivity contributes to low viremia in most HBeAg-negative patients. , 2007, Gastroenterology.

[41]  Kook Hwan Kim,et al.  Hepatitis B virus X protein induces hepatic steatosis via transcriptional activation of SREBP1 and PPARgamma. , 2007, Gastroenterology.

[42]  R. Norel,et al.  cDNA microarray analysis of HBV transgenic mouse liver identifies genes in lipid biosynthetic and growth control pathways affected by HBV , 2005, Journal of medical virology.

[43]  M. Westphal,et al.  Oncoretrovirus and Lentivirus Vectors Pseudotyped with Lymphocytic Choriomeningitis Virus Glycoprotein: Generation, Concentration, and Broad Host Range , 2002, Journal of Virology.

[44]  L. Moore,et al.  A regulatory cascade of the nuclear receptors FXR, SHP-1, and LRH-1 represses bile acid biosynthesis. , 2000, Molecular cell.

[45]  T. A. Kerr,et al.  Molecular basis for feedback regulation of bile acid synthesis by nuclear receptors. , 2000, Molecular cell.

[46]  D. Trono,et al.  A Third-Generation Lentivirus Vector with a Conditional Packaging System , 1998, Journal of Virology.

[47]  J. Marin,et al.  Pathophysiological and pharmacological implications of elucidating the molecular bases of the interaction between HBV and the bile acid transporter NTCP. , 2015, Annals of hepatology.

[48]  M. Anwer,et al.  Sodium-dependent bile salt transporters of the SLC10A transporter family: more than solute transporters , 2013, Pflügers Archiv - European Journal of Physiology.

[49]  B. Stieger.  The role of the sodium-taurocholate cotransporting polypeptide (NTCP) and of the bile salt export pump (BSEP) in physiology and pathophysiology of bile formation. , 2011, Handbook of experimental pharmacology.

[50]  S. Strom,et al.  Chimeric mice with humanized liver: tools for the study of drug metabolism, excretion, and toxicity. , 2010, Methods in molecular biology.