Severe acute respiratory syndrome coronavirus (SARS-CoV) is a recently emerged pathogen. To predict and control the evolutionary and pathogenic potential of SARS-CoV, a better understanding of the basic biology and pathogenesis should lead to the development of vaccines and therapies. We have developed a full length infectious cDNA for mouse hepatitis virus (MHV), transmissible gastroenteritis virus (TGEV) and,SARS-CoV. The overall hypothesis in this proposal is that deletion or rearrangement of selected ORFs and/or """"""""rewiring """""""" of the coronavirus transcription network will identify virulence alleles, attenuate SARS-CoV pathogenesis in vivo, stabilize against recombination and reversion, and result in candidate vaccines that protect against in vivo challenge. Attenuated viruses developed during the course of these studies will also serve as seed stocks for safe, killed vaccines.
In aim 1, Dr. Baric tests that hypothesis that the SARS-CoV group specific genes encode luxury functions in vitro. Two SARS-CoV group specific ORFs encode IFN antagonists that likely influence pathogenesis (see Palese, Project 3) and viable viruses have been isolated that contain deletions in these ORFs. We will delete systematically knockout the SARS-CoV group specific ORFs and characterize the phenotype of rescued viruses. The goal is to isolate replication competent viruses that lack group specific ORFs, are attenuated and protect against wildtype challenge in vivo.
In aim 2, Dr. Baric tests the hypothesis that the SARS-CoV gene order rearranged viruses recombine less frequently than wildtype viruses and are attenuated in animal models.
In aim 3, Dr. Baric tests the hypothesis that the transcriptional regulatory sequences of SARS-CoV can be rewired and that these mutants will be highly resistant to recombination with wildtype and other coronaviruses. The goal is to create a genome platform that can 1) house a suite of attenuated alleles and 2) contain genetic traps that impart lethal genome instabilities, triggered after recombining with other coronaviruses.
In aim 4, the in vivo pathogenesis of these live attenuated and select killed viruses are compared in mice and ferrets. We will also determine if select mutants induce immune responses that protect against wildtype challenge. The project simultaneously increases our understanding of the molecular biology of SARS-CoV replication and transcription, the molecular mechanisms governing coronavirus recombination and strategies to impede RNA recombination repair of candidate vaccine strains, and the evolutionary potential of the SARS-CoV genome as an emerging human pathogen.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Program Projects (P01)
Project #
5P01AI059443-03
Application #
7341635
Study Section
Special Emphasis Panel (ZAI1)
Project Start
Project End
Budget Start
2007-02-01
Budget End
2008-01-31
Support Year
3
Fiscal Year
2007
Total Cost
$259,764
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
DUNS #
608195277
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Gonzalez, P N; Pavlicev, M; Mitteroecker, P et al. (2016) Genetic structure of phenotypic robustness in the collaborative cross mouse diallel panel. J Evol Biol 29:1737-51
Sheahan, Timothy; Whitmore, Alan; Long, Kristin et al. (2011) Successful vaccination strategies that protect aged mice from lethal challenge from influenza virus and heterologous severe acute respiratory syndrome coronavirus. J Virol 85:217-30
Eckerle, Lance D; Becker, Michelle M; Halpin, Rebecca A et al. (2010) Infidelity of SARS-CoV Nsp14-exonuclease mutant virus replication is revealed by complete genome sequencing. PLoS Pathog 6:e1000896
Brooke, Christopher B; Deming, Damon J; Whitmore, Alan C et al. (2010) T cells facilitate recovery from Venezuelan equine encephalitis virus-induced encephalomyelitis in the absence of antibody. J Virol 84:4556-68
Li, Kelvin; Venter, Eli; Yooseph, Shibu et al. (2010) ANDES: Statistical tools for the ANalyses of DEep Sequencing. BMC Res Notes 3:199
Rockx, Barry; Donaldson, Eric; Frieman, Matthew et al. (2010) Escape from human monoclonal antibody neutralization affects in vitro and in vivo fitness of severe acute respiratory syndrome coronavirus. J Infect Dis 201:946-55
Frieman, Matthew B; Chen, Jun; Morrison, Thomas E et al. (2010) SARS-CoV pathogenesis is regulated by a STAT1 dependent but a type I, II and III interferon receptor independent mechanism. PLoS Pathog 6:e1000849
Zornetzer, Gregory A; Frieman, Matthew B; Rosenzweig, Elizabeth et al. (2010) Transcriptomic analysis reveals a mechanism for a prefibrotic phenotype in STAT1 knockout mice during severe acute respiratory syndrome coronavirus infection. J Virol 84:11297-309
Frieman, Matthew; Ratia, Kiira; Johnston, Robert E et al. (2009) Severe acute respiratory syndrome coronavirus papain-like protease ubiquitin-like domain and catalytic domain regulate antagonism of IRF3 and NF-kappaB signaling. J Virol 83:6689-705
Rockx, Barry; Baas, Tracey; Zornetzer, Gregory A et al. (2009) Early upregulation of acute respiratory distress syndrome-associated cytokines promotes lethal disease in an aged-mouse model of severe acute respiratory syndrome coronavirus infection. J Virol 83:7062-74

Showing the most recent 10 out of 37 publications