DN-200434

Earlier, we have reported a series of inverse agonist of estrogen related receptor gamma (ERRγ), viz DN200434, GSK5182 and its derivates, that enhance sodium iodide (Na+/I-) symporter (NIS) function through mitogen-activated protein (MAP) kinase signaling in anaplastic thyroid cancer cells. But, the effect of our recently discovered DN200434, on NIS function in papillary thyroid cancer (PTC) refractory to radioactive iodine (RAI) therapy has not been reported. Herein, we studied the effects of DN200434 in RAI-resistant PTC cells on ERRγ-mediated regulation of NIS function.

RAI-refractory BCPAP cells were exposed to lower concentrations of DN200434 and the NIS function in the PTC cells was serially assessed by radioiodine uptake assay. Immunoblot assay was performed to study the effect of DN200434 on ERRγ, NIS, the mitogen-activated protein (MAP) kinase pathway, and iodide-handling genes. To examine whether the DN200434-induced NIS functional activity can be affected by inhibition of the MAP kinase pathway, radioiodine uptake was performed after the application of U0126, a selective MEK inhibitor to DN200434-treated cells. Finally, the cytotoxic effect of 131I was determined in DN200434 treated and untreated cells by clonogenic assay.

Treatment of DN200434 resulted in dose-dependent increases in iodide uptake in BCPAP cells, downregulation of ERRγ protein and the activation of extracellular signal-regulated kinase (ERK) 1/2. Treatment of the specific MEK inhibitor overturned the increased radioiodine uptake and ERK1/2 activation of BCPAP cells. DN200434 treatment enhanced the membrane localization of NIS in BCPAP cells. Clonogenic assay results revealed enhanced cytotoxic effects of 131I in DN200434 pre-exposed BCPAP cells.

Thus, we successfully demonstrate that our novel inverse agonist of ERRγ, DN200434, enhances the responsiveness of radioiodine therapy by modulating NIS function in RAI-refractory papillary thyroid cancer cells via the regulation of ERRγ and the MAP kinase signaling pathway.

Sorafenib, originally identified as an inhibitor of multiple oncogenic kinases, induces ferroptosis in hepatocellular carcinoma (HCC) cells. Several pathways that mitigate sorafenib-induced ferroptosis confer drug resistance; thus strategies that enhance ferroptosis increase sorafenib efficacy. Orphan nuclear receptor estrogen-related receptor γ (ERRγ) is upregulated in human HCC tissues and plays a role in cancer cell proliferation. The aim of this study was to determine whether inhibition of ERRγ with DN200434, an orally available inverse agonist, can overcome resistance to sorafenib through induction of ferroptosis. Sorafenib-resistant HCC cells were less sensitive to sorafenibinduced ferroptosis and showed significantly higher ERRγ levels than sorafenib-sensitive HCC cells. DN200434 induced lipid peroxidation and ferroptosis in sorafenib-resistant HCC cells. Mechanistically, DN200434 increased mitochondrial ROS generation by reducing glutathione/glutathione disulfide levels, which subsequently reduced mTOR activity and GPX4 levels. DN200434 induced amplification of the antitumor effects of sorafenib was confirmed in a tumor xenograft model. The present results indicate that DN200434 may be a novel therapeutic strategy to re-sensitize HCC cells to sorafenib. [BMB Reports 2022; 55(11): 547-552].