Hepatitis E virus (HEV) is an important but extremely understudied pathogen. Genotypes 1 and 2 are restricted to humans, whereas genotypes 3 and 4 infect humans and other animals. The long-term goals of this project are to delineate the mechanisms of HEV cross-species infection and identify host and viral determinants of host range and pathogenesis.
In specific aim 1, we hypothesize that the hypervariable region (HVR) in ORF1 and the inserted host gene sequences in HVR determine HEV host range. Our preliminary studies suggested that HVR and its adjacent region are involved in HEV tropism. The recent findings that host S17 and S19 gene sequences were inserted into the HVR of HEV that resulted in recombinant viruses with expanded cell tropism of different animal origins lends further credence to our hypothesis. We will determine (a) if a genotype 1 human HEV (infecting humans only) will acquire the ability to infect pigs and rabbits if its HVR is replaced with that o a zoonotic genotype 3 HEV, (b) if the genotype 1 human HEV with an inserted host S17 sequence in the HVR will acquire the ability to infect pigs and rabbits, and (c) if the adjacent X domain in ORF1 plays a role, together with HVR, in determining the host range.
In specific aim 2, we hypothesize that the genotype- specific HVR sequences and/or inserted host sequences contribute to HEV pathogenicity. Increased pathogenicity by recombination events between host gene sequence and virus genomes has been reported for other viruses. The fact that the neurological and chronic cases of hepatitis E are all caused by the zoonotic genotype 3 HEV and that quasispecies compartmentalization in CSF and serum of clonal HEV sequences within the HVR region was identified from neurological cases suggested that the HVR plays a role in virus pathogenicity and emergence of neurotropic variants. We will determine (a) if the genotype 3 virus with the S17 insertion in HVR is more virulent than the virus without the insertion, (b) if the genotype 3 virus with a S17 insertion establishes chronic infection in pigs, and (c) if the HVR and its adjacent regions evolve during chronic HEV infection in pigs to produce neurotropic variants.
In specific aim 3, we hypothesize that interferon stimulated gene 15 (ISG15) plays a role in HEV infection and host anti-HEV response. Our preliminary studies showed that HEV replication requires an active ubiquitin-proteasome system and that overexpression of ISG15 inhibited HEV replication in a HEV replicon system. The findings that the papain-like cysteine protease (PCP) domain in HEV ORF1 has deubiquitinating activity and carries out deISGylation of ISG15-conjugated proteins support our hypothesis. By using an efficient HEV infection cell culture model with genotype 3 HEV Kernow C- 1 strain, we will determine (a) if overexpression of ISG15 will inhibit genotype 3 HEV infection, and if so, what is the mechanism of inhibition, (b) if HEV counteracts ISG15 production and ISGylation, and if so, if the PCP domain plays a role in antagonizing ISG15 function. By completing this project, we expect to identify the viral determinant(s) for HEV host range and pathogenicity and delineate the role of ISG15 in anti-HEV response.

Public Health Relevance

Hepatitis E virus (HEV) is an important but extremely understudied human pathogen causing significant public health problem in developing countries but is also endemic in the United States and other industrialized countries. In this project, we wil identify the genetic determinant(s) for HEV cross-species infection and host range, and delineate the role of interferon-stimulated gene 15 (ISG15) in HEV infection and host anti-HEV defense. The information from this project will be important for devising effective prevention and treatment strategies against HEV.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI074667-06
Application #
8689884
Study Section
Hepatobiliary Pathophysiology Study Section (HBPP)
Program Officer
Koshy, Rajen
Project Start
2007-06-01
Project End
2017-06-30
Budget Start
2014-07-01
Budget End
2015-06-30
Support Year
6
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Virginia Polytechnic Institute and State University
Department
Veterinary Sciences
Type
Schools of Veterinary Medicine
DUNS #
City
Blacksburg
State
VA
Country
United States
Zip Code
24060
Kenney, Scott P; Wentworth, Jacquelyn L; Heffron, Connie L et al. (2015) Replacement of the hepatitis E virus ORF3 protein PxxP motif with heterologous late domain motifs affects virus release via interaction with TSG101. Virology 486:198-208
Kenney, Scott P; Meng, Xiang-Jin (2015) Identification and fine mapping of nuclear and nucleolar localization signals within the human ribosomal protein S17. PLoS One 10:e0124396
Kenney, Scott P; Meng, Xiang-Jin (2015) Therapeutic targets for the treatment of hepatitis E virus infection. Expert Opin Ther Targets 19:1245-60
Kenney, Scott P; Meng, Xiang-Jin (2015) The lysine residues within the human ribosomal protein S17 sequence naturally inserted into the viral nonstructural protein of a unique strain of hepatitis E virus are important for enhanced virus replication. J Virol 89:3793-803
Yugo, Danielle M; Cossaboom, Caitlin M; Meng, Xiang-Jin (2014) Naturally occurring animal models of human hepatitis E virus infection. ILAR J 55:187-99
Yugo, Danielle M; Meng, Xiang-Jin (2013) Hepatitis E virus: foodborne, waterborne and zoonotic transmission. Int J Environ Res Public Health 10:4507-33
Sanford, B J; Emerson, S U; Purcell, R H et al. (2013) Serological evidence for a hepatitis e virus-related agent in goats in the United States. Transbound Emerg Dis 60:538-45
Cossaboom, Caitlin M; Cordoba, Laura; Sanford, Brenton J et al. (2012) Cross-species infection of pigs with a novel rabbit, but not rat, strain of hepatitis E virus isolated in the United States. J Gen Virol 93:1687-95
Meng, X J (2012) Emerging and re-emerging swine viruses. Transbound Emerg Dis 59 Suppl 1:85-102
Kenney, Scott P; Pudupakam, R S; Huang, Yao-Wei et al. (2012) The PSAP motif within the ORF3 protein of an avian strain of the hepatitis E virus is not critical for viral infectivity in vivo but plays a role in virus release. J Virol 86:5637-46

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