FH535 suppresses osteosarcoma growth in vitro and inhibits wnt signaling through tankyrases

Osteosarcoma (OS) is an aggressive primary bone tumor which exhibits aberrantly activated Wnt signaling. The canonical Wnt signaling cascade has been shown to drive cancer progression and metastasis through the activation of β-catenin. Hence, small molecule inhibitors of Wnt targets are being explored as primary or adjuvant chemotherapy. In this study, we have investigated the ability of FH₅₃₅, an antagonist of Wnt signaling, to inhibit the growth of OS cells. We found that FH₅₃₅\ was cytotoxic in all OS cell lines which were tested (143b, U₂OS, SaOS-₂, HOS, K₇M₂) but well tolerated by normal human osteoblast cells. Additionally, we have developed an in vitro model of doxorubicin-resistant OS and found that these cells were highly responsive to FH₅₃₅ treatment. Our analysis provided evidence that FH₅₃₅ strongly inhibited markers of canonical Wnt signaling. In addition, our findings demonstrate a reduction in PAR-modification of Axin2 indicating inhibition of the tankyrase 1/2 enzymes. Moreover, we observed inhibition of auto-modification of PARP1 in the presence of FH₅₃₅, indicating inhibition of PARP1 enzymatic activity. These data provide evidence that FH₅₃₅ acts through the tankyrase 1/2 enzymes to suppress Wnt signaling and could be explored as a potent chemotherapeutic agent for the control of OS.