Stephan Urban，海德堡，德国——HBV相关实验室及人物介绍系列

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1 简要信息：

Stephan Urban
Head of Hepatitis B Research Group
Development of envelope protein-derived entry inhibitors for the treatment of acute and chronic HBV and HDV infection
主页：
http://www.klinikum.uni-heidelberg.de/AG-Urban.104952.0.html

2 简介：
他是HepRG细胞株的发现者之一，原始文献是：
Gripon, P., et al. , Infection of a human hepatoma cell line by hepatitis B virus. Proc Natl Acad Sci U S A, 2002. 99(24): p. 15655-60.

3 课题方向：
Characterization of viral and cellular determinants essential for HBV entry

Unlike other pathogenic viruses, the initial steps of the hepatitis B virus (HBV) infection, including attachment and receptor recognition, but also subsequent processes, such as the endocytosis pathway, the mechanism of membrane fusion or the intracellular transport events are still unknown. The objectives of this project comprise the deciphering and functional characterization of viral and cellular components that participate in the establishment of the viral infection. A particular focus is related to the identification of host cell factors that function as primary attachment receptors. In this respect we scrutinize the dependence of HBV binding and infection on cell-surface associated glycosaminoglycans. In addition to the visualization of HBV-entry using fluorescently labeld HBV-particles we are trying to dissect the dependence of intracellular sorting and trafficking events on the host cell cytoskeleton.

Identification and analysis of molecular components involved DHBV entry

Early steps in the duck hepatitis B virus (DHBV) life cycle were the subject of many studies in the past. This led to the identification of one receptor component, duck carboxypeptidase D (dCPD/gp180). Although a crucial function for this glycoprotein at some stage of DHBV entry is well established, some features (e.g. preferential TGN localization, binding to the most inner, transmembrane adjacent C-domain, lack of liver specific expression) question the role of CPD as a simple attachment receptor. In this project we aim to elucidate the exact role of CPD during viral binding to hepatocytes and e.g. fusion of viral and cellular membranes. Moreover we are trying to identify components other than dCPD which participate in viral entry. Finally, we are interested in the resolution of the structure of the dCPD domain interacting with the large DHBV surface protein.

Ultrastructural dynamics of Hepatitis B Virus maturation

In the infected cell, newly synthesized genomic RNA is packaged together with the viral Pol-protein and a cellular chaperone complex into capsids. The RNA is then reverse transcribed into a partially double-stranded DNA within the capsid. Under natural conditions, almost exclusively mature capsids containing DNA are enveloped to be secreted as virions. The selectivity of this process depends on a still undefined maturation signal that communicates the RNA-DNA-transition from the interior to the capsid exterior.

The large envelope protein of HBV (L-protein) adopts two transmembrane topologies: At first, its preS-region is located on the cytosolic side of the ER-membrane and acts as a matrix-domain that binds the mature capsid to enable its envelopment. The same region is thereafter required on the viron surface as a receptor binding domain to initiate the next round of infection. So far, it is unknown if the dual topology is already pre-formed prior to viral budding and therefore stably present in the viral particles. Alternatively, the topological switch could proceed mainly in free virions after secretion as a second, distinct maturation step. Using highly purified viral particles, we are characterising the determinants and dynamics of the two proposed maturation steps with biochemical and imaging techniques.

Development of novel entry inhibitors for the treatment of acute and chronic Hepatitis B and Hepatitis D virus infections

Specific inhibition of virus entry into host cells is an established and powerful therapeutic concept to come across acute infections. For chronic infections e.g. HIV, Enfurvitide/Fuzeon®, a peptide inhibitor which prevents fusion of the viral membrane with the lymphocyte membrane has successfully been approved recently. The remarkable in vitro and in vivo inhibitory activity of acylated HBV-preS-derived peptides implies possible clinical applications like prophylaxis of HBV-reinfection after liver transplantation or the protection of newborns from vertical HBV-transmission from their infected mothers. Since it is still unclear to which degree the maintenance of a chronic HBV carrier state depends on constantly ongoing new infection events in the liver (whilst simultaneously infected cells are eliminated by the immune system), entry inhibition might also be an intriguing concept for the treatment of chronic HBV infections.

We are currently performing preclinical studies including the improvement of knowledge on production, efficacy, stability, bioavailability of HBV envelope protein-derived lipopeptides. We are aiming to optimize their in vivo application form and investigate to what extend the immunologic responses contribute to the therapeutic outcome. Aim of these studies is the subsequent accomplishment of a Phase I clinical trial in the near future.

4 代表论文
详细参见：http://www.klinikum.uni-heidelbe ... tions.110121.0.html

Urban S.New insights into hepatitis B and hepatitis delta virus entry.Future Virology, May 2008, Vol. 3, No. 3, Pages 253-264

Petersen J, Dandri M, Mier W, Lütgehetmann M, Volz T, von Weizsäcker F, Haberkorn U, Fischer L, Pollok JM, Erbes B, Seitz S, Urban S.
Prevention of hepatitis B virus infection in vivo by entry inhibitors derived from the large envelope protein.
Nat Biotechnol. 2008 Mar;26(3):335-41. Epub 2008 Feb 24

Schulze A, Gripon P, Urban S.
Hepatitis B virus infection initiates with a large surface protein-dependent binding to heparan sulfate proteoglycans.
Hepatology. 2007 Dec;46(6):1759-68.


PS：
这是悬赏活动之“HBV相关实验室及人物介绍系列”模板，具体参见：
http://bbs.virology.com.cn/thread-7594-1-1.html

发表于 2009-2-21 07:33 Bigben

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Zhang, Z. and B. Zehnder, et al. (2016). "Visualization of Hepatitis B Virus Entry: Novel Tools and Approaches to Directly  Follow Virus Entry into Hepatocytes." FEBS Lett.
Blank, A. and C. Markert, et al. (2016). "First-in-human application of the first-in-class hepatitis B and hepatitis D virus entry inhibitor myrcludex B." J Hepatol.
Bogomolov, P. and A. Alexandrov, et al. (2016). "Treatment of chronic hepatitis D with the entry inhibitor myrcludex B - first results of a Phase Ib/IIa study." J Hepatol.
Li, W. and S. Urban (2016). "Entry of hepatitis B and hepatitis D virus into hepatocytes: Basic insights and clinical implications." J Hepatol 64(1 Suppl): S32-40.
Uhl, P. and F. Helm, et al. (2016). "A liposomal formulation for the oral application of the investigational hepatitis B drug Myrcludex B." Eur J Pharm Biopharm 103: 159-66.
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Lempp, F. A. and P. Mutz, et al. (2016). "Evidence that hepatitis B virus replication in mouse cells is limited by the lack of a host cell dependency factor." J Hepatol 64(3): 556-64.
Urban, S. (2016). "Liver capsule: Entry and entry inhibition of hepatitis B virus and hepatitis delta virus into hepatocytes." Hepatology 63(2): 633.
Liang, T. J. and T. M. Block, et al. (2015). "Present and future therapies of hepatitis B: From discovery to cure." Hepatology 62(6): 1893-908.
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Lempp, F. A. and S. Urban (2014). "Inhibitors of hepatitis B virus attachment and entry." Intervirology 57(3-4): 151-7.
Oehler, N. and T. Volz, et al. (2014). "Binding of hepatitis B virus to its cellular receptor alters the expression profile of genes of bile acid metabolism." Hepatology 60(5): 1483-93.
Watashi, K. and S. Urban, et al. (2014). "NTCP and beyond: opening the door to unveil hepatitis B virus entry." Int J Mol Sci 15(2): 2892-905.
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Meier, A. and S. Mehrle, et al. (2013). "Myristoylated PreS1-domain of the hepatitis B virus L-protein mediates specific binding to differentiated hepatocytes." Hepatology 58(1): 31-42.
Schulze, A. and K. Mills, et al. (2012). "Hepatocyte polarization is essential for the productive entry of the hepatitis B  virus." Hepatology 55(2): 373-83.
Lutgehetmann, M. and L. V. Mancke, et al. (2012). "Humanized chimeric uPA mouse model for the study of hepatitis B and D virus interactions and preclinical drug evaluation." Hepatology 55(3): 685-94.
von Hahn, T. and A. Schulze, et al. (2011). "The novel immunosuppressive protein kinase C inhibitor sotrastaurin has no pro-viral effects on the replication cycle of hepatitis B or C virus." PLoS One 6(9): e24142.
Schieck, A. and T. Muller, et al. (2010). "Solid-phase synthesis of the lipopeptide Myr-HBVpreS/2-78, a hepatitis B virus entry inhibitor." Molecules 15(7): 4773-83.
Ni, Y. and J. Sonnabend, et al. (2010). "The pre-s2 domain of the hepatitis B virus is dispensable for infectivity but serves a spacer function for L-protein-connected virus assembly." J Virol 84(8): 3879-88.
Urban, S. and A. Schulze, et al. (2010). "The replication cycle of hepatitis B virus." J Hepatol 52(2): 282-4.
Schulze, A. and A. Schieck, et al. (2010). "Fine mapping of pre-S sequence requirements for hepatitis B virus large envelope  protein-mediated receptor interaction." J Virol 84(4): 1989-2000.
Kim, D. H. and Y. Ni, et al. (2008). "An anti-viral peptide derived from the preS1 surface protein of hepatitis B virus." BMB Rep 41(9): 640-4.
Gudima, S. and Y. He, et al. (2008). "Primary human hepatocytes are susceptible to infection by hepatitis delta virus assembled with envelope proteins of woodchuck hepatitis virus." J Virol 82(15): 7276-83.
Petersen, J. and M. Dandri, et al. (2008). "Prevention of hepatitis B virus infection in vivo by entry inhibitors derived from the large envelope protein." Nat Biotechnol 26(3): 335-41.
Bohne, F. and M. Chmielewski, et al. (2008). "T cells redirected against hepatitis B virus surface proteins eliminate infected  hepatocytes." Gastroenterology 134(1): 239-47.
Tavakoli, S. and I. Mederacke, et al. (2008). "Peripheral blood dendritic cells are phenotypically and functionally intact in chronic hepatitis B virus (HBV) infection." Clin Exp Immunol 151(1): 61-70.
Shin, E. C. and U. Seifert, et al. (2007). "Proteasome activator and antigen-processing aminopeptidases are regulated by virus-induced type I interferon in the hepatitis C virus-infected liver." J Interferon Cytokine Res 27(12): 985-90.
Schulze, A. and P. Gripon, et al. (2007). "Hepatitis B virus infection initiates with a large surface protein-dependent binding to heparan sulfate proteoglycans." Hepatology 46(6): 1759-68.
Seitz, S. and S. Urban, et al. (2007). "Cryo-electron microscopy of hepatitis B virions reveals variability in envelope capsid interactions." EMBO J 26(18): 4160-7.
Engelke, M. and K. Mills, et al. (2006). "Characterization of a hepatitis B and hepatitis delta virus receptor binding site." Hepatology 43(4): 750-60.
Glebe, D. and S. Urban, et al. (2005). "Mapping of the hepatitis B virus attachment site by use of infection-inhibiting preS1 lipopeptides and tupaia hepatocytes." Gastroenterology 129(1): 234-45.
Gripon, P. and I. Cannie, et al. (2005). "Efficient inhibition of hepatitis B virus infection by acylated peptides derived  from the large viral surface protein." J Virol 79(3): 1613-22.
Urban, S. (2004). "Binding of duck carboxypeptidase D to duck hepatitis B virus." Methods Mol Med 95: 199-212.
Gripon, P. and S. Rumin, et al. (2002). "Infection of a human hepatoma cell line by hepatitis B virus." Proc Natl Acad Sci U S A 99(24): 15655-60.
Urban, S. and P. Gripon (2002). "Inhibition of duck hepatitis B virus infection by a myristoylated pre-S peptide of the large viral surface protein." J Virol 76(4): 1986-90.
Urban, S. and S. Urban, et al. (2001). "Efficient pyrophosphorolysis by a hepatitis B virus polymerase may be a primer-unblocking mechanism." Proc Natl Acad Sci U S A 98(9): 4984-9.
Breiner, K. M. and S. Urban, et al. (2001). "Envelope protein-mediated down-regulation of hepatitis B virus receptor in infected hepatocytes." J Virol 75(1): 143-50.
Urban, S. and D. L. Tyrrell (2000). "An in vitro system for the enzymological analysis of avian hepatitis B virus replication and inhibition in core particles." Antiviral Res 45(3): 185-97.
Urban, S. and C. Schwarz, et al. (2000). "Receptor recognition by a hepatitis B virus reveals a novel mode of high affinity virus-receptor interaction." EMBO J 19(6): 1217-27.
Urban, S. and C. Kruse, et al. (1999). "A soluble form of the avian hepatitis B virus receptor. Biochemical characterization and functional analysis of the receptor ligand complex." J Biol Chem 274(9): 5707-15.
Poussin, K. and H. Dienes, et al. (1999). "Expression of mutated hepatitis B virus X genes in human hepatocellular carcinomas." Int J Cancer 80(4): 497-505.
Sirma, H. and R. Weil, et al. (1998). "Cytosol is the prime compartment of hepatitis B virus X protein where it colocalizes with the proteasome." Oncogene 16(16): 2051-63.
Breiner, K. M. and S. Urban, et al. (1998). "Carboxypeptidase D (gp180), a Golgi-resident protein, functions in the attachment and entry of avian hepatitis B viruses." J Virol 72(10): 8098-104.
Urban, S. and K. M. Breiner, et al. (1998). "Avian hepatitis B virus infection is initiated by the interaction of a distinct pre-S subdomain with the cellular receptor gp180." J Virol 72(10): 8089-97.
Urban, S. and E. Hildt, et al. (1997). "Isolation and molecular characterization of hepatitis B virus X-protein from a baculovirus expression system." Hepatology 26(4): 1045-53.
Hildt, E. and S. Urban, et al. (1996). "Isolation of highly purified, functional carboxy-terminally truncated hepatitis B virus middle surface protein activators from eucaryotic expression systems." Hepatology 24(3): 502-7.