Figure 6. Alisertib decreases virus replication.

in vitro but enhances HSV1716 replication and persistence in vivo. A. S462TY tumor cells were treated with alisertib (80 nM) or vehicle for 48 hours and then infected with HSV1716 at the various MOI indicated. Cells were harvested at 24, 48 and 72 hours post virus injection and analyzed for virus replication by plaque assay. For the in vitro replication assays, all data points represent mean +SD, (n = 4 replicates per MOI at each time point). C. and D. S462TY or SK-N-AS tumor cells were treated with alisertib (80 nM) or vehicle for 48 hours and then infected with HSV1716-GFP (MOI 0.1). Green fluorescence was monitored over time using the Incucyte. For the in vitro cytotoxicity assays, all data points represent mean ± SD, (n = 4 measurements per well). All the experiments were repeated 3 times. Representative data are shown. E. Mice bearing SK-N-AS xenograft tumors were treated with alisertib (20 mg/kg twice daily) or vehicle and given a single intratumoral injection of 1 × 10⁷ pfu HSV1716, and the amount of infectious virus recovered at various times determined by plaque assay. For the in vivo replication studies, all data points represent mean ± SD, (n = 3-4 replicates per time point).