%0 Journal Article
%T Modulation of GSK-3¦Â Activity in Venezuelan Equine Encephalitis Virus Infection
%A Kylene Kehn-Hall
%A Aarthi Narayanan
%A Lindsay Lundberg
%A Gavin Sampey
%A Chelsea Pinkham
%A Irene Guendel
%A Rachel Van Duyne
%A Svetlana Senina
%A Kimberly L. Schultz
%A Eric Stavale
%A M. Javad Aman
%A Charles Bailey
%A Fatah Kashanchi
%J PLOS ONE
%D 2012
%I Public Library of Science (PLoS)
%R 10.1371/journal.pone.0034761
%X Alphaviruses, including Venezuelan Equine Encephalitis Virus (VEEV), cause disease in both equine and humans that exhibit overt encephalitis in a significant percentage of cases. Features of the host immune response and tissue-specific responses may contribute to fatal outcomes as well as the development of encephalitis. It has previously been shown that VEEV infection of mice induces transcription of pro-inflammatory cytokines genes (e.g., IFN-¦Ã, IL-6, IL-12, iNOS and TNF-¦Á) within 6 h. GSK-3¦Â is a host protein that is known to modulate pro-inflammatory gene expression and has been a therapeutic target in neurodegenerative disorders such as Alzheimer's. Hence inhibition of GSK-3¦Â in the context of encephalitic viral infections has been useful in a neuroprotective capacity. Small molecule GSK-3¦Â inhibitors and GSK-3¦Â siRNA experiments indicated that GSK-3¦Â was important for VEEV replication. Thirty-eight second generation BIO derivatives were tested and BIOder was found to be the most potent inhibitor, with an IC50 of ~0.5 ¦ÌM and a CC50 of >100 ¦ÌM. BIOder was a more potent inhibitor of GSK-3¦Â than BIO, as demonstrated through in vitro kinase assays from uninfected and infected cells. Size exclusion chromatography experiments demonstrated that GSK-3¦Â is found in three distinct complexes in VEEV infected cells, whereas GSK-3¦Â is only present in one complex in uninfected cells. Cells treated with BIOder demonstrated an increase in the anti-apoptotic gene, survivin, and a decrease in the pro-apoptotic gene, BID, suggesting that modulation of pro- and anti-apoptotic genes contributes to the protective effect of BIOder treatment. Finally, BIOder partially protected mice from VEEV induced mortality. Our studies demonstrate the utility of GSK-3¦Â inhibitors for modulating VEEV infection.
%U http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0034761