Kinome profiling upon HCMV infection using the MIB-MS approach

Human cytomegalovirus (HCMV) is a significant cause of disease in immune-compromised adults and immune naïve newborns. No vaccine exists to prevent HCMV infection, and current antiviral therapies have toxic side effects that limit the duration and intensity of their use. There is thus an urgent need for new strategies to treat HCMV and repurposing existing drugs as antivirals is an attractive approach. Virus-induced changes in infected cells are often driven by changes in cellular kinase activity, leading us to hypothesize that defining the complement of kinases (the kinome), whose activity or expression is altered during infection would identify existing kinase inhibitors that could be repurposed as new antivirals. To this end, we applied a recently developed technique, MIB-MS kinome profiling, to quantitatively measure perturbations in >240 cellular kinases simultaneously in cells infected with a laboratory-adapted (AD169) or clinical (TB40E) HCMV strain using a label-free approach. Significant time-dependent changes for multiple kinases including cell cycle, receptor tyrosine and mitotic kinases were observed. Based on these data, we tested antiviral activity of 14 kinase inhibitors, and the most potent inhibitor blocked HCMV early gene expression and viral DNA accumulation. These results show the utility of kinome profiling to screen kinase inhibitors that can be repurposed as antivirals.