title: HSV activates Akt to trigger calcium release and promote viral entry: novel candidate target for treatment and suppression.
abstract: HSV triggers intracellular calcium release to promote viral entry. We hypothesized that Akt signaling induces the calcium responses and contributes to HSV entry. Exposure of human cervical and primary genital tract epithelial, neuronal, or keratinocyte cells to HSV serotype 2 resulted in rapid phosphorylation of Akt. Silencing of Akt with small interfering RNA prevented the calcium responses, blocked viral entry, and inhibited plaque formation by 90% compared to control siRNA. Susceptibility to infection was partially restored if Akt was reintroduced into silenced cells with an Akt-expressing plasmid. HSV-2 variants deleted in glycoproteins B or D failed to induce Akt phosphorylation, and coimmunoprecipitation studies indicated that Akt interacts with glycoprotein B. Cell-surface expression of Akt was rapidly induced in response to HSV exposure. Miltefosine (50 μM), a licensed drug that blocks Akt phosphorylation, inhibited HSV-induced calcium release, viral entry, and plaque formation following infection with acyclovir-sensitive and resistant clinical isolates. Miltefosine blocked amplification of HSV from explanted ganglia to epithelial cells; viral yields were significantly less in miltefosine compared to control-treated cocultures (P<0.01). Together, these findings identify a novel role for Akt in viral entry, link Akt and calcium signaling, and suggest a new target for HSV treatment and suppression.
authors.affiliation: Department of Pediatrics, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461, USA.
authors.forename: Natalia
authors.initials: N
authors.lastname: Cheshenko
authors.forename: Janie B
authors.initials: JB
authors.lastname: Trepanier
authors.forename: Martha
authors.initials: M
authors.lastname: Stefanidou
authors.forename: Niall
authors.initials: N
authors.lastname: Buckley
authors.forename: Pablo
authors.initials: P
authors.lastname: Gonzalez
authors.forename: William
authors.initials: W
authors.lastname: Jacobs
authors.forename: Betsy C
authors.initials: BC
authors.lastname: Herold
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology; D005260:Female; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism; D018258:Herpesvirus 2, Human / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D054884:Host-Pathogen Interactions; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism / Q000821:virology; D018613:Microscopy, Confocal; D009154:Mutation; D010766:Phosphorylation / Q000187:drug effects; D010767:Phosphorylcholine / Q000031:analogs & derivatives / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D053586:Virus Internalization
chemical_list: D000970:Antineoplastic Agents; D006023:Glycoproteins; D014764:Viral Proteins; D010767:Phosphorylcholine; C039128:miltefosine; D051057:Proto-Oncogene Proteins c-akt; D002118:Calcium
keywords: genital herpes; glycoprotein B; glycoprotein D; miltefosine