OKN-007 is an Effective Anticancer Therapeutic Agent Targeting Inflammatory and Immune Metabolism Pathways in Endometrial Cancer.

Advanced-stage endometrial cancer patients typically receive a combination of platinum and paclitaxel chemotherapy. However, limited treatment options are available for those with recurrent disease, and there is a need to identify alternative treatment options for the advanced setting. Our goal was to evaluate the pre-clinical efficacy and mechanism of action of Oklahoma Nitrone 007 (OKN-007) alone and in combination with carboplatin and paclitaxel in endometrial cancer. The effect of OKN-007 on the metabolic viability of endometrial cancer cells in both two- and three-dimensional (2D and 3D) cultures, as well as on clonogenic growth, in vitro was assessed. We also evaluated OKN-007 in vivo using an intraperitoneal xenograft model and targeted gene expression profiling to determine the molecular mechanism and gene expression programs altered by OKN-007. Our results showed that endometrial cancer cells were generally sensitive to OKN-007 in both 2D and 3D cultures. OKN-007 displayed a reduction in 3D spheroid and clonogenic growth. Subsequent targeted gene expression profiling revealed that OKN-007 significantly downregulated the immunosuppressive and immunometabolic enzyme indolamine 2,3-dioxygenase 1 (IDO1) (-11.27-fold change) and modulated upstream inflammatory pathways that regulate IDO1 expression (interferon- (IFN-), Jak-STAT, TGF-ß, and NF-kB), downstream IDO1 effector pathways (mTOR and aryl hydrocarbon receptor (AhR)) and altered T-cell co-signaling pathways. OKN-007 treatment reduced IDO1, SULF2, and TGF-ß protein expression in vivo, and inhibited TGF-ß, NF-kB, and AhR- receptor-mediated nuclear signaling in vitro. These findings indicate that OKN-007 surmounts pro-inflammatory, immunosuppressive, and pro-tumorigenic pathways and is a promising approach for the effective treat endometrial cancer. Significance Statement Women with advanced and recurrent endometrial cancer have limited therapeutic options. OKN-007, which has minimal toxicity and is currently being evaluated in early-phase clinical trials for the treatment of cancer, is a potential new strategy for the treatment of endometrial cancer.

Targeting doublecortin-like kinase 1 reveals a novel strategy to circumvent chemoresistance and metastasis in ovarian cancer.

Ovarian cancer (OvCa) has a dismal prognosis because of its late-stage diagnosis and the emergence of chemoresistance. Doublecortin-like kinase 1 (DCLK1) is a serine/threonine kinase known to regulate cancer cell "stemness", epithelial-mesenchymal transition (EMT), and drug resistance. Here we show that DCLK1 is a druggable target that promotes chemoresistance and tumor progression of high-grade serous OvCa (HGSOC). Importantly, high DCLK1 expression significantly correlates with poor overall and progression-free survival in OvCa patients treated with platinum chemotherapy. DCLK1 expression was elevated in a subset of HGSOC cell lines in adherent (2D) and spheroid (3D) cultures, and the expression was further increased in cisplatin-resistant (CPR) spheroids relative to their sensitive controls. Using cisplatin-sensitive and resistant isogenic cell lines, pharmacologic inhibition (DCLK1-IN-1), and genetic manipulation, we demonstrate that DCLK1 inhibition was effective at re-sensitizing cells to cisplatin, reducing cell proliferation, migration, and invasion. Using kinase domain mutants, we demonstrate that DCLK1 kinase activity is critical for mediating CPR. The combination of cisplatin and DCLK1-IN-1 showed a synergistic cytotoxic effect against OvCa cells in 3D conditions. Targeted gene expression profiling revealed that DCLK1 inhibition in CPR OvCa spheroids significantly reduced TGFß signaling, and EMT. We show in vivo efficacy of combined DCLK1 inhibition and cisplatin in significantly reducing tumor metastases. Our study shows that DCLK1 is a relevant target in OvCa and combined targeting of DCLK1 in combination with existing chemotherapy could be a novel therapeutic approach to overcome resistance and prevent OvCa recurrence.

Quercetin inhibits human metastatic ovarian cancer cell growth and modulates components of the intrinsic apoptotic pathway in PA-1 cell line.

Ovarian cancer is the leading cause of gynaecology related cancer death worldwide. It is often diagnosed with an advanced stage. Apoptosis is a process of programmed cell death controlled by cell cycle machinery and several signaling pathways. Plant-derived compounds have received an increased interest in the treatment of cancer. Quercetin is a flavonoid present in fruits and vegetables which possess anticancer properties. Several studies have been demonstrated that quercetin induces apoptosis in various cancers. However, the apoptotic role of quercetin in metastatic ovarian cancer has not been extensively studied. In the present study, we investigated the apoptotic effect of quercetin on human metastatic ovarian cancer PA-1 cell line. Quercetin treatment (0-200 µM) for 24h decreases PA-1 cells viability in a dose-dependent manner. The effective dose was identified as 50 and 75 µM based on MTT assay. Quercetin induces apoptosis in metastatic ovarian cancer cells which were confirmed by AO/EtBr dual staining, DAPI staining and DNA fragmentation assay. Molecules involved in the intrinsic apoptotic pathway were altered by quercetin. Interestingly, antiapoptotic molecules such as Bcl-2, Bcl-xL were decreased while proapoptotic molecules such as caspase-3, caspase-9, Bid, Bad, Bax and cytochrome c were increased in the quercetin-treated PA-1 cells. Our results indicate that quercetin induces mitochondrial-mediated apoptotic pathway and thus it inhibits the growth of metastatic ovarian cancer cells.

Ameliorative effect of α-tocopherol on polychlorinated biphenyl (PCBs) induced testicular Sertoli cell dysfunction in F1 prepuberal rats.

The study was conducted to investigate the protective role of α-tocopherol against polychlorinated biphenyls (PCBs) induced effect in Sertoli cell function of F1 prepuberal rats. Dams were grouped into six; each group consists of six animals. Group 1-control treated with corn oil as vehicle, group II- 0.5mgPCBs/kg bw/day, group III- 0.5mgPCBs/kg bw/day with α-tocopherol (50mg α-tocopherol/kg bw/day), group IV- 1mgPCBs/kg bwt/day, group V- 1mgPCBs/kg bw/day with α-tocopherol (50mg α-tocopherol/kg bw/day) and group VI - α-tocopherol alone treated orally from postpartum day1-20. Male offspring rats were euthanized on post natal day 21. Testes were collected for the histological examination and Sertoli cell isolation. The protein levels of follicle-stimulating hormone receptor, androgen binding protein, androgen receptor, estrogen receptor α & ß, Inhibin-ß, transferrin, claudin-11, occludin, E-cadherin, connexin-43, c-fos, c-jun, SF1, USF1 & 2 were studied using western blot method. The testicular architecture was affected in the PCBs exposed rats but this effect was restored by α-tocopherol supplementation. PCBs decreased the protein levels of FSHR, AR, ABP, ERα & ß, transferrin, claudin-11, occludin, E-cadherin, connexin-43, c-fos, c-jun, SF1, USF1 & 2 whereas inhibin-ß protein level was found to be increased in Sertoli cells. These results suggested that α-tocopherol has ameliorative role against PCBs induced testicular Sertoli cell dysfunction in F1 progeny.