Ricolinostat enhances adavosertib-induced mitotic catastrophe in TP53-mutated head and neck squamous cell carcinoma cells

TP53 mutation is one of the most frequent gene mutations in head and neck squamous cell carcinoma (HNSCC) and could be a potential therapeutic target. Recently, the WEE1 G2 checkpoint kinase (WEE1) inhibitor adavosertib (Adv) has attracted attention because of its selective cytotoxicity against TP53-mutated cells and has shown promising activity in early phase clinical trials. In the present study, it was demonstrated that combined treatment with Adv and a selective histone deacetylase 6 (HDAC6) inhibitor, ricolinostat (RCS), synergistically enhanced cell death induction in four out of five HNSCC cell lines with TP53 mutation (CAL27, SAS, HSC-3, and OSC-19), one HNSCC cell line with impaired TP53 function by HPV-infection (UPCI-SCC154), and TP53-knockout human lung cancer cell line (A549 TP53-KO), but not in TP53 wild-type A549 cells. Time-lapse imaging showed that RCS enhanced the Adv-induced mitotic catastrophe. Consistent with this, RCS treatment suppressed checkpoint kinase 1 (Chk1) (Ser345) phosphorylation and co-administration of RCS with Adv suppressed cyclin-dependent kinase 1 (Tyr15) phosphorylation along with increased expression of γ-H2A.X, a marker of DNA double-strand breaks in CAL27 cells. These data showed that RCS enhanced Adv-induced premature mitotic entry and cell death induction in the mitotic phase. However, although HDAC6 knockdown enhanced Adv-induced cell death with γ-H2A.X elevation, HDAC6 knockdown did not repress Chk1 phosphorylation in CAL27 cells. Our data demonstrated that the co-administration of RCS with Adv in HNSCC cells resulted in the suppression of Chk1 activity, leading to synergistically enhanced apoptosis via mitotic catastrophe in a p53-dependent manner. This enhanced cell death appeared to be partially mediated by the inhibition of HDAC6 activity by RCS.