Supplementary materials Material and Methods Supplemental Figure 1: Quantitative assessment of mRNA expression of Mcl-1, RelA and Stat3 in anaplastic thyroid carcinoma (ATC) cell lines and resected ATC's using qRT-PCR. Supplemental Figure 2: Expressions of NF�B-p65 and Stat3pY in ATC. Supplemental Figure 3: Assessment of medical and staining parameters relationship to the overall survival (OS) in the ATC cohort. Supplemental Figure 4: Meta analysis of previously published expression profiles on ATC's. Supplemental Figure 5: Model schematic displaying drug synergy from the drug combination of sorafenib/quinacrine through the targeting of expression of anti-apoptotic Mcl-1. Supplemental Figure 6: In vivo optical near-infrared (NIR) imaging of orthotopic xenografts of human ATC cells. Supplemental Figure 7: Serum chemistry indicates lack of apparent toxicity by the S/Q combination. Supplemental Figure 8: Hematology indicates lack of apparent toxicity by the S/Q combination. Supplemental Figure 9: Reduced surface occupancy of CD34-positive small vessel endothelium and VEGF-expression in 8505C tumor xenografts treated with the combination of sorafenib and quinacrine.
ARTICLE ABSTRACT
Purpose and Experimental Design: Anaplastic thyroid cancer (ATC) comprises approximately 2% of all thyroid cancers, and its median survival rate remains poor. It is responsible for more than one third of thyroid cancer–related deaths. ATC is frequently resistant to conventional therapy, and NFκB signaling has been proposed to be a feature of the disease. We aimed to assess the activity of the antimalaria drug quinacrine known to target NFκB signaling in combination with the clinically relevant kinase inhibitor sorafenib in ATC cells. The presence of NFκB-p65/RELA and its target MCL1 was demonstrated in ATC by meta-data gene set enrichment analysis and IHC. We assessed the responses of a panel of human ATC cell lines to quinacrine and sorafenib in vitro and in vivo.Results: We detected increased expression of NFκB-p65/RELA and MCL1 in the nucleus of a subset of ATC compared with non-neoplastic thyroid. ATC cells were found to respond with additive/synergistic tumor cell killing to the combination of sorafenib plus quinacrine in vitro, and the drug combination improves survival of immunodeficient mice injected orthotopically with ATC cells as compared with mice administered either compound alone or doxorubicin. We also demonstrate that the combination of sorafenib and quinacrine is well tolerated in mice. At the molecular level, quinacrine and sorafenib inhibited expression of prosurvival MCL1, pSTAT3, and dampened NFκB signaling.Conclusions: The combination of quinacrine and sorafenib targets emerging molecular hallmarks of ATC and shows promising results in clinically relevant models for the disease. Further testing of sorafenib plus quinacrine can be conducted in ATC patients. Clin Cancer Res; 22(24); 6192–203. ©2016 AACR.
