Topical microbicides that block transmission of herpes simplex virus (HSV) and other sexually transmitted pathogens are urgently needed. The observation that heparan sulfate proteoglycans serve as attachment receptors propelled the development of sulfonated polymers, which are now in clinical trial, as candidate microbicides. Identification of new agents that target steps post-binding requires elucidation, at a molecular level, of the signaling pathways required for fusion of the viral envelope with the cell plasma membrane and nuclear transport of viral capsids. Preliminary studies indicate that viral invasion requires activation of calcium (Ca2+) signaling and tyrosine phosphorylation of focal adhesion kinase (FAK) and other cellular proteins. Activation of Ca2+ and phosphorylation signaling pathways are common mechanisms associated with invasion by other microbes, most notably HIV. The studies proposed in this application build on these observations. Focusing on HSV serotype 2 (HSV-2), the predominant cause of genital herpes, these studies will examine the cellular signaling pathways required for viral invasion and the role viral envelope glycoproteins play in this process. Most studies of viral entry have been conducted using permanent non-polarized epithelial cell lines. However, the polarity of epithelial cells is fundamental to their structure and function and influences susceptibility to infection. Preliminary work indicates that HSV- 2 infection of polarized epithelial cells is more efficient following apical compared to basolateral exposure. Therefore, the mechanism of viral invasion in polarized cell culture systems will also be explored. Observations in polarized cells will more accurately reflect the conditions in vivo. Results of these studies will enhance understanding of the molecular biology of HSV invasion and should facilitate the rational design of new topical antimicrobials to prevent HSV transmission and acquisition.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
7R01AI061679-04
Application #
7574877
Study Section
Virology - A Study Section (VIRA)
Program Officer
Beisel, Christopher E
Project Start
2005-02-15
Project End
2010-01-31
Budget Start
2007-10-01
Budget End
2008-01-31
Support Year
4
Fiscal Year
2007
Total Cost
$293,206
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Pediatrics
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Cheshenko, Natalia; Trepanier, Janie B; González, Pablo A et al. (2014) Herpes simplex virus type 2 glycoprotein H interacts with integrin ?v?3 to facilitate viral entry and calcium signaling in human genital tract epithelial cells. J Virol 88:10026-38
Cheshenko, Natalia; Trepanier, Janie B; Stefanidou, Martha et al. (2013) HSV activates Akt to trigger calcium release and promote viral entry: novel candidate target for treatment and suppression. FASEB J 27:2584-99
Stefanidou, Martha; Ramos, Irene; Mas Casullo, Veronica et al. (2013) Herpes simplex virus 2 (HSV-2) prevents dendritic cell maturation, induces apoptosis, and triggers release of proinflammatory cytokines: potential links to HSV-HIV synergy. J Virol 87:1443-53
Holm, Christian K; Jensen, Søren B; Jakobsen, Martin R et al. (2012) Virus-cell fusion as a trigger of innate immunity dependent on the adaptor STING. Nat Immunol 13:737-43
Cheshenko, Natalia; Trepanier, Janie B; Segarra, Theodore J et al. (2010) HSV usurps eukaryotic initiation factor 3 subunit M for viral protein translation: novel prevention target. PLoS One 5:e11829
Madan, Rebecca Pellett; Mesquita, Pedro M M; Cheshenko, Natalia et al. (2007) Molecular umbrellas: a novel class of candidate topical microbicides to prevent human immunodeficiency virus and herpes simplex virus infections. J Virol 81:7636-46
Cheshenko, Natalia; Liu, Wen; Satlin, Lisa M et al. (2007) Multiple receptor interactions trigger release of membrane and intracellular calcium stores critical for herpes simplex virus entry. Mol Biol Cell 18:3119-30
Galen, Benjamin; Cheshenko, Natalia; Tuyama, Ana et al. (2006) Access to nectin favors herpes simplex virus infection at the apical surface of polarized human epithelial cells. J Virol 80:12209-18
Cheshenko, Natalia; Liu, Wen; Satlin, Lisa M et al. (2005) Focal adhesion kinase plays a pivotal role in herpes simplex virus entry. J Biol Chem 280:31116-25