Few therapy options exist for patients with advanced papillary and anaplastic thyroid cancer. We and others have previously identified c-Src as a key mediator of thyroid cancer pro-tumorigenic processes and a promising therapeutic target for thyroid cancer. To increase the efficacy of targeting Src in the clinic, we sought to define mechanisms of resistance to the Src inhibitor, dasatinib, to identify key pathways to target in combination. Using a panel of thyroid cancer cell lines expressing clinically relevant mutations in BRAF or RAS, which were previously developed to be resistant to dasatinib, we identified a switch to a more invasive phenotype in the BRAF-mutant cells as a potential therapy escape mechanism. This phenotype switch is driven by FAK kinase activity, and signaling through the p130Cas>c-Jun signaling axis. We have further shown this more invasive phenotype is accompanied by alterations in the secretome through the increased expression of pro-inflammatory cytokines, including IL-1β, and the pro-invasive metalloprotease, MMP-9. Furthermore, IL-1β signals via a feedforward autocrine loop to promote invasion through a FAK>p130Cas>c-Jun>MMP-9 signaling axis. We further demonstrate that upfront combined inhibition of FAK and Src synergistically inhibits growth and invasion, and induces apoptosis in a panel of BRAF- and RAS-mutant thyroid cancer cell lines. Together our data demonstrate that acquired resistance to single-agent Src inhibition promotes a more invasive phenotype through an IL-1β>FAK>p130Cas>c-Jun >MMP signaling axis, and that combined inhibition of FAK and Src has the potential to block this inhibitor-induced phenotype switch.
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We thank Dr. Christopher Korch, UCCC, and Randall Wong at the B. Davis Center BioResources Core Facility for STR profiling of the cell lines. We thank Dr. Rytis Prekeris for assistance with the zymography assays. This work was supported by NIH/NCI grant 1RO1CA164193 (RES), 1R01CA222299 (RES), American Cancer Society RSG-13-060-01-TBE (RES), the Cancer League of Colorado (RES), and the Front Range Cancer Challenge Fellowship (KMM). The UCCC DNA Sequencing, Flow Cytometry, and Protein Production/Mab/Tissue Culture Shared Resources are supported by NCI Cancer Center support grant, P30CA046934.
Conflict of interest
The authors declare that they have no conflict of interest.
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Kessler, B.E., Mishall, K.M., Kellett, M.D. et al. Resistance to Src inhibition alters the BRAF-mutant tumor secretome to promote an invasive phenotype and therapeutic escape through a FAK>p130Cas>c-Jun signaling axis. Oncogene 38, 2565–2579 (2019). https://doi.org/10.1038/s41388-018-0617-1
ITK inhibition induced in vitro and in vivo anti-tumor activity through downregulating TCR signaling pathway in malignant T cell lymphoma
Cancer Cell International (2019)
Resistance to Src inhibition alters the BRAF-mutant tumor secretome to promote an invasive phenotype and therapeutic escape through a FAK>p130Cas>c-Jun signaling axis