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Targeting host kinases for the treatment of herpes keratitis
Author(s) -
ALEKSEEV O,
JENNINGS S,
AZIZKHANCLIFFORD J
Publication year - 2012
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2012.t112.x
Subject(s) - corneal epithelium , biology , in vivo , keratitis , cell culture , virology , epithelium , kinase , in vitro , wortmannin , cancer research , microbiology and biotechnology , genetics , phosphatidylinositol
Purpose Herpes keratitis (HK) is a common cause of blindness in the developed world. A large number of HK cases are refractory to antiviral agents and ultimately result in permanent corneal damage. The purpose of this study is to identify new therapeutic targets against HSV and to assess their antiviral potential in tissue culture experiments, as well as in more sophisticated models of HK. Specifically, we focused on the involvement of ataxia telangiectasia mutated (ATM) and its downstream target Chk2 in facilitating productive HSV infection in corneal epithelium. Methods We used human corneal epithelial cell lines – hTCEpi and HCE – as in vitro models of corneal HSV infection, and also developed an ex vivo model of acute corneal epithelial infection, where explanted human and rabbit corneoscleral buttons were infected and maintained in organ culture. We are currently utilizing an in vivo mouse model of ocular herpes to further validate our findings. Small molecule inhibitors of ATM (KU‐55933, wortmannin, caffeine) and Chk2 (Chk2 inhibitor II), as well as RNAi against ATM and Chk2, were used to inhibit these two kinases. We used plaque assays and qPCR to assess the infectious particle production, genome replication, and transcriptional activity of HSV in corneal epithelium. Results Small molecule or RNAi‐mediated inhibition of ATM or Chk2 greatly suppressed the replication and transcription of the viral genome, as well as the infectious particle production. This was observed in the tissue culture models and, importantly, in organotypically explanted human and rabbit corneas. Results of the animal studies will also be presented. Conclusion This study identifies two host kinases – ATM and Chk2 – as potential novel therapeutic targets against herpes keratitis.