: Chronic HCV infection is a major threat to the public health. Current therapies have limited efficacy, but the search for more effective treatments is hampered by the lack of available animal models of HCV infection. The chimpanzee (Pan troglodytes) is the only animal species permissive for infection with this virus. This deficiency will be addressed by developing a small nonhuman primate model of hepatitis C involving the closely related, unclassified flavivirus GBV-B. GBV-B replicates to high titers, is hepatotropic, and causes liver disease in susceptible tamarins (Saquinus sp.). Since tamarins are more readily available than chimpanzees for such studies, GBV-B infection of these animals represents a potentially useful surrogate for studies of hepatitis C. However, although GBV-B among all animal viruses has the closest phylogenetic relationship to HCV, its proteins still share only approximately 25% identity at the amino acid level. Moreover, unlike HCV, GBV-B does not appear capable of establishing persistent infection in these animals. These features of GBV- B limit its usefulness. To overcome these limitations, the applicants will construct chimeric genome-length GBV-B cDNA clones in which specific functional domains of HCV are inserted in lieu of homologous GBV-B sequence. The hypothesis is that the close phylogenetic relationship between GBV-B and HCV will allow the rescue of viable chimeric viruses from these clones, and that these viruses will represent uniquely valuable resources to the research community since they will allow the in vivo evaluation of candidate inhibitors of critical HCV replication functions in a readily available and relatively inexpensive small, nonhuman primate species.
Under Aim 1, the investigators have constructed a fulllength, infectious cDNA copy of the GBV-B genome. The infectivity of RNA transcribed from this clone has been demonstrated following intrahepatic injection of the RNA in a susceptible tamarin.
In Aim 2, the investigators are constructing infectious chimeric flavivirus cDNAs containing the following HCV domains within a GBV-B background: the internal ribosome entry site (IRES), the major proteinase (NS3) with its cofactor molecule (NS4A), the RNA helicase (NS3) and the RNA dependent, RNA polymerase (NSSB).
In Aim 3, chimeras in which the structural proteins of GBV-B and HCV are placed within the genetic background of the alternate virus will be constructed. For both Aims 2 and 3, the applicants will assess the ability of RNAs transcribed from these chimeric cDNA clones to induce infection in tamarins following intrahepatic inoculation, and determine the extent to which the viruses rescued from these clones cause acute or chronic liver disease on subsequent passage in these nonhuman primates.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Resource-Related Research Projects (R24)
Project #
1R24RR015081-01A2
Application #
6436180
Study Section
National Center for Research Resources Initial Review Group (RIRG)
Program Officer
O'Neill, Raymond R
Project Start
2002-02-15
Project End
2006-01-31
Budget Start
2002-02-15
Budget End
2003-01-31
Support Year
1
Fiscal Year
2002
Total Cost
$285,452
Indirect Cost
Name
University of Texas Medical Br Galveston
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041367053
City
Galveston
State
TX
Country
United States
Zip Code
77555
Benureau, Yann; Warter, Lucile; Malcolm, Bruce A et al. (2010) A comparative analysis of the substrate permissiveness of HCV and GBV-B NS3/4A proteases reveals genetic evidence for an interaction with NS4B protein during genome replication. Virology 406:228-40
Weatherford, Trudie; Chavez, Deborah; Brasky, Kathleen M et al. (2009) Lack of adaptation of chimeric GB virus B/hepatitis C virus in the marmoset model: possible effects of bottleneck. J Virol 83:8062-75
Warter, Lucile; Cohen, Lisette; Benureau, Yann et al. (2009) A cooperative interaction between nontranslated RNA sequences and NS5A protein promotes in vivo fitness of a chimeric hepatitis C/GB virus B. PLoS One 4:e4419
Chen, Zihong; Benureau, Yann; Rijnbrand, Rene et al. (2007) GB virus B disrupts RIG-I signaling by NS3/4A-mediated cleavage of the adaptor protein MAVS. J Virol 81:964-76
Chevalier, Christophe; Saulnier, Aure; Benureau, Yann et al. (2007) Inhibition of hepatitis C virus infection in cell culture by small interfering RNAs. Mol Ther 15:1452-62
Targett-Adams, Paul; Schaller, Torsten; Hope, Graham et al. (2006) Signal peptide peptidase cleavage of GB virus B core protein is required for productive infection in vivo. J Biol Chem 281:29221-7
Rijnbrand, Rene; Yang, Yan; Beales, Lucy et al. (2005) A chimeric GB virus B with 5' nontranslated RNA sequence from hepatitis C virus causes hepatitis in tamarins. Hepatology 41:986-94
Rijnbrand, Rene; Thiviyanathan, Varatharasa; Kaluarachchi, Kumaralal et al. (2004) Mutational and structural analysis of stem-loop IIIC of the hepatitis C virus and GB virus B internal ribosome entry sites. J Mol Biol 343:805-17
Martin, Annette; Bodola, Francis; Sangar, David V et al. (2003) Chronic hepatitis associated with GB virus B persistence in a tamarin after intrahepatic inoculation of synthetic viral RNA. Proc Natl Acad Sci U S A 100:9962-7