Efficacy of adavosertib therapy against anaplastic thyroid cancer

in Endocrine-Related Cancer
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  • 1 Department of Internal Medicine, New Taipei Municipal TuCheng Hospital, New Taipei City, Taiwan
  • 2 Department of Internal Medicine, Chang Gung Memorial Hospital, Taoyuan, Taiwan
  • 3 Chang Gung University, Taoyuan, Taiwan
  • 4 Center for Big Data Analytics and Statistics, Chang Gung Memorial Hospital, Taoyuan, Taiwan
  • 5 Laboratory of Preclinical Pharmacology Core, Memorial Sloan-Kettering Cancer Center, New York, New York, USA
  • 6 Department of Surgery, Memorial Sloan-Kettering Cancer Center, New York, New York, USA

Correspondence should be addressed to S-F Lin: mmg@cgmh.org.tw

(T-C Chou is now at PD Science, LLC., Paramus, New Jersey, USA)

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Wee1 is a kinase that regulates the G2/M progression by the inhibition of CDK1, which is critical for ensuring DNA damage repair before initiation of mitotic entry. Targeting Wee1 may be a potential strategy in the treatment of anaplastic thyroid cancer, a rare but lethal disease. The therapeutic effects of adavosertib, a Wee1 inhibitor for anaplastic thyroid cancer was evaluated in this study. Adavosertib inhibited cell growth in three anaplastic thyroid cancer cell lines in a dose-dependent manner. Cell cycle analysis revealed cells were accumulated in the G2/M phase. Adavosertib induced caspase-3 activity and led to apoptosis. Adavosertib monotherapy showed significant retardation of the growth of two anaplastic thyroid cancer tumor models. The combination of adavosertib with dabrafenib and trametinib revealed strong synergism in vitro and demonstrated robust suppression of tumor growth in vivo in anaplastic thyroid cancer xenograft models with BRAF V600E mutation. The combination of adavosertib with either sorafenib or lenvatinib also demonstrated synergism in vitro and had strong inhibition of tumor growth in vivo in an anaplastic thyroid cancer xenograft model. No appreciable toxicity appeared in mice treated with either a single agent or combination treatment. Our findings suggest adavosertib holds the promise for the treatment of patients with anaplastic thyroid cancer.

Supplementary Materials

    • Supplementary Figure 1. The effects of adavosertib on cell cycle distribution in ATC cells. Cell cycle analysis measuring the DNA content of 1 x 104 events for each sample using flow cytometry was performed in ATC cells treated with placebo or adavosertib (500 nmol/L) for 72 hours (8505C) and 48 hours (8305C and KAT18).
    • Supplementary Figure 2. The effects of adavosertib on cleaved PARP expression in ATC cell lines. ATC cells were plated at 1 x 106 cells in 100-mm Petri dishes in 10 mL media overnight and treated with placebo or adavosertib (500 nmol/L) for indicated periods. The levels of cleaved PARP were evaluated by immunoblot as described in Figure 3F. Band densities were imaged and quantified using Molecular Imager VersaDoc MP 4000 system (Bio-Rad). The ratios of cleaved PARP to β-actin at each time point were calculated. Relative expression was calculated using control cells as a reference for each cell line. Adavosertib treatment induced the expression of cleaved PARP by 24 hours (KAT18) and 48 hours (8505C and 8305C).
    • Supplementary Figure 3. Dabrafenib and trametinib treatment decreased cell viability in ATC cell lines. LDH assays were performed to determine cell viability by dabrafenib and trametinib treatment for a 4-day treatment course in ATC cell lines. Dose-response curves were obtained for dabrafenib (A) and trametinib (B) for 8505C and 8305C cell lines. The median-effect dose (IC50) profiles of dabrafenib (C) and trametinib (D) in ATC cell lines were determined by CompuSyn software.
    • Supplementary Figure 4. The combination of adavosertib and dabrafenib plus trametinib suppresses tumor growth in four individual 8305C xenografts. Four nude mice with established 8305C flank tumors were treated with oral gavage of triple combination of adavosertib (50 mg/kg), dabrafenib (30 mg/kg) and trametinib (0.6 mg/kg) once a day for three cycles of 5-day-on and 2-day-off therapy as described in Figure 5D. Triple combination therapy decreased tumor growth in all mice during the treatment period (Day 0-21). After discontinuation of treatment, the triple therapy was enduring with complete response in 50% (2 of 4) of mice until day 35 when this study was closed. Black arrow, triple combination treatment.
    • Supplementary Figure 5. The molecular effects of adavosertib therapy in 8505C and 8305C tumors. Tumor levels of p-CDK1 (Tyr15), p-CHK1 (Ser345), p-H2AX (Ser139), cleaved caspase-3, cleaved PARP, PCNA and p-ERK1/2 (Thr202/Tyr204) were evaluated in 8505C and 8305C xenografts after a single oral dosing of adavosertib (50 mg/kg) using Western blot.
    • Supplementary Figure 6. Sorafenib and lenvatinib treatment decreased cell viability in ATC cell lines. LDH assays were performed to determine cell viability by sorafenib and lenvatinib treatment for a 4-day treatment course in three ATC cell lines. Dose-response curves were obtained for sorafenib (A) and lenvatinib (B) for each ATC cell lines. The median-effect dose (IC50) profiles of sorafenib (C) and lenvatinib (D) treatment in ATC cell lines were determined by CompuSyn software.

 

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