azd-6244 and Ovarian-Neoplasms

Low-grade serous carcinoma of the ovary is chemoresistant but mutations in the MAPK pathway could be targeted to control tumour growth. We therefore assessed the safety and activity of selumetinib, an inhibitor of MEK1/2, for patients with this cancer.. In this open-label, single-arm phase 2 study, women (aged ≥18 years) with recurrent low-grade serous ovarian or peritoneal carcinoma were given selumetinib (50 mg twice daily, orally) until progression. The primary endpoint was the proportion of patients who had an objective tumour response according to RECIST version 1.1, assessed for all the treated patients. Analysis was by intention to treat. This study is registered with ClinicalTrials.gov, number NCT00551070.. 52 patients were enrolled between Dec 17, 2007, and Nov 23, 2009. All were eligible for analyses. Eight (15%) patients had an objective response to treatment-one patient had a complete response and seven had partial responses. 34 (65%) patients had stable disease. There were no treatment-related deaths. Grade 4 toxicities were cardiac (one), pain (one), and pulmonary events (one). Grade 3 toxicities that occurred in more than one patient were gastrointestinal (13), dermatological (nine), metabolic (seven), fatigue (six), anaemia (four), pain (four), constitutional (three), and cardiac events (two).. Selumetinib is well tolerated, and is active in the treatment of recurrent low-grade serous carcinoma of the ovary or peritoneum. The findings suggest that inhibitors of the MAPK pathway warrant further investigation in these patients.. National Cancer Institute.

Topics: Adult; Aged; Benzimidazoles; Cystadenocarcinoma, Serous; Female; Humans; MAP Kinase Signaling System; Middle Aged; Mitogen-Activated Protein Kinase Kinases; Mutation; Neoplasm Recurrence, Local; Ovarian Neoplasms; Peritoneal Neoplasms; Proto-Oncogene Proteins; Proto-Oncogene Proteins B-raf; Proto-Oncogene Proteins p21(ras); ras Proteins

Topics: Adenocarcinoma, Clear Cell; Animals; Antineoplastic Combined Chemotherapy Protocols; Apoptosis; Benzimidazoles; Benzoxazoles; Biomarkers, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; Drug Synergism; Female; Gene Expression Regulation, Neoplastic; Humans; Indazoles; Mice; Mice, Nude; Morpholines; Ovarian Neoplasms; Phosphatidylinositol 3-Kinases; Phosphorylation; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-akt; Pyrimidines; Sulfonamides; TOR Serine-Threonine Kinases; Tumor Cells, Cultured; Xenograft Model Antitumor Assays

Although 67% of high-grade serous ovarian cancers (HGSOC) express the estrogen receptor (ER), most fail antiestrogen therapy. Because MAPK activation is frequent in ovarian cancer, we investigated if estrogen regulates MAPK and if MEK inhibition (MEKi) reverses antiestrogen resistance.. Effects of MEKi (selumetinib), antiestrogen (fulvestrant), or both were assayed in ER-positive HGSOC in vitro and in xenografts. Response biomarkers were investigated by gene expression microarray and reverse phase protein array (RPPA). Genes differentially expressed in two independent primary HGSOC datasets with high versus low pMAPK by RPPA were used to generate a "MAPK-activated gene signature." Gene signature components that were reversed by MEKi were then identified.. High intratumor pMAPK independently predicts decreased survival (HR, 1.7; CI > 95%,1.3-2.2; P = 0.0009) in 408 HGSOC from The Cancer Genome Atlas. A differentially expressed "MAPK-activated" gene subset was also prognostic. "MAPK-activated genes" in HGSOC differ from those in breast cancer. Combined MEK and ER blockade showed greater antitumor effects in xenografts than monotherapy. Gene set enrichment analysis and RPPA showed that dual therapy downregulated DNA replication and cell-cycle drivers, and upregulated lysosomal gene sets. Selumetinib reversed expression of a subset of "MAPK-activated genes" in vitro and/or in xenografts. Three of these genes were prognostic for poor survival (P = 0.000265) and warrant testing as a signature predictive of MEKi response.. High pMAPK is independently prognostic and may underlie antiestrogen failure. Data support further evaluation of fulvestrant and selumetinib in ER-positive HGSOC. The MAPK-activated HGSOC signature may help identify MEK inhibitor responsive tumors.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; Benzimidazoles; Drug Resistance, Neoplasm; Drug Synergism; Enzyme Activation; Estradiol; Estrogen Receptor Modulators; Female; Fulvestrant; Humans; Kaplan-Meier Estimate; MAP Kinase Signaling System; Mice, Inbred NOD; Mice, SCID; Mitogen-Activated Protein Kinases; Neoplasms, Cystic, Mucinous, and Serous; Ovarian Neoplasms; Receptors, Estrogen; Transcriptome; Treatment Outcome; Xenograft Model Antitumor Assays

No effective systemic therapy exists for patients with metastatic low-grade serous (LGS) ovarian cancers. BRAF and KRAS mutations are common in serous borderline (SB) and LGS ovarian cancers, and MEK inhibition has been shown to induce tumor regression in a minority of patients; however, no correlation has been observed between mutation status and clinical response. With the goal of identifying biomarkers of sensitivity to MEK inhibitor treatment, we performed an outlier analysis of a patient who experienced a complete, durable, and ongoing (> 5 years) response to selumetinib, a non-ATP competitive MEK inhibitor.. Next-generation sequencing was used to analyze this patient's tumor as well as an additional 28 SB/LGS tumors. Functional characterization of an identified novel alteration of interest was performed.. Analysis of the extraordinary responder's tumor identified a 15-nucleotide deletion in the negative regulatory helix of the MAP2K1 gene encoding for MEK1. Functional characterization demonstrated that this mutant induced extracellular signal-regulated kinase pathway activation, promoted anchorage-independent growth and tumor formation in mice, and retained sensitivity to selumetinib. Analysis of additional LGS/SB tumors identified mutations predicted to induce extracellular signal-regulated kinase pathway activation in 82% (23 of 28), including two patients with BRAF fusions, one of whom achieved an ongoing complete response to MEK inhibitor-based combination therapy.. Alterations affecting the mitogen-activated protein kinase pathway are present in the majority of patients with LGS ovarian cancer. Next-generation sequencing analysis revealed deletions and fusions that are not detected by older sequencing approaches. These findings, coupled with the observation that a subset of patients with recurrent LGS ovarian cancer experienced dramatic and durable responses to MEK inhibitor therapy, support additional clinical studies of MEK inhibitors in this disease.

Topics: Animals; Benzimidazoles; Biomarkers, Tumor; Carcinoma, Ovarian Epithelial; Cystadenocarcinoma, Serous; Drug Resistance, Neoplasm; Female; Follow-Up Studies; High-Throughput Nucleotide Sequencing; Humans; MAP Kinase Kinase 1; Mice; Middle Aged; Mutation; Neoplasm Grading; Neoplasms, Glandular and Epithelial; NIH 3T3 Cells; Ovarian Neoplasms; Prognosis; Tumor Stem Cell Assay

Ovarian cancer is a silent disease with a poor prognosis that urgently requires new therapeutic strategies. In low-grade ovarian tumours, mutations in the MAP3K BRAF gene constitutively activate the downstream kinase MEK. Here we demonstrate that an additional MAP3K, MAP3K8 (TPL-2/COT), accumulates in high-grade serous ovarian carcinomas (HGSCs) and is a potential prognostic marker for these tumours. By combining analyses on HGSC patient cohorts, ovarian cancer cells and patient-derived xenografts, we demonstrate that MAP3K8 controls cancer cell proliferation and migration by regulating key players in G1/S transition and adhesion dynamics. In addition, we show that the MEK pathway is the main pathway involved in mediating MAP3K8 function, and that MAP3K8 exhibits a reliable predictive value for the effectiveness of MEK inhibitor treatment. Our data highlight key roles for MAP3K8 in HGSC and indicate that MEK inhibitors could be a useful treatment strategy, in combination with conventional chemotherapy, for this disease.

Topics: Animals; Antineoplastic Agents; Benzimidazoles; Biomarkers, Tumor; Carcinogenesis; Cell Line, Tumor; Cystadenocarcinoma, Serous; Female; Humans; MAP Kinase Kinase Kinases; MAP Kinase Signaling System; Mice; Mice, Nude; Mitogen-Activated Protein Kinase Kinases; Ovarian Neoplasms; Prognosis; Proto-Oncogene Proteins; Ribosomal Protein S6 Kinases, 90-kDa; Xenograft Model Antitumor Assays

To investigate the effect of phosphatidylinositol 3 kinase (PI3K) inhibitor LY294002 combined with mitogen activated protein kinase kinase (MEK) inhibitor AZD6244 on the proliferation of cisplatin-resistant SKOV3/DDP ovarian cancer cell line.. The alteration in the cell proliferation of SKOV3/DDP cells treated with 5, 10, 20, 40, 80 μmol/L LY294002 and 1, 2, 4, 8, 16 μmol/L AZD6244 alone or together, was detected by MTT assay. The 50% inhibitory concentration (IC50) and combined index (CI) were calculated. The concentration of the combination was obtained based on the MTT assay, and the cells were divided into four groups: the control group, LY294002 group (5 μmol/L), AZD6244 group (7 μmol/L) and combination group (LY294002 5 μmol/L and AZD6244 7 μmol/L); Forty-eight hours later, MTT assay was used to detect the cell proliferation and calculate cell doubling time; colony formation assay was performed to detect colony formation efficiency; Annexin V-PE/7-ADD staining combined with flow cytometry (FCM) was used to detect the apoptotic rates and cell cycle; Western blotting was employed to detect the levels of AKT, phosphorylated AKT (p-AKT), extracellular signal-regulated kinase 1/2 (ERK1/2), phosphorylated ERK1/2 (p-ERK1/2), cyclin D1 and cleaved caspase-3 protein.. Cell growth was inhibited by LY294002 or AZD6244 alone, and the effect was strengthened when LY294002 was combined with AZD6244 (CI<1), indicating that the two inhibitors showed synergistic effect. The proliferation of cells was significantly slower, the cell doubling time was significantly prolonged, and colony number was reduced in the combined treatment group (P<0.01) compared with the single inhibitor treatment group and the control group. Meanwhile, FCM demonstrated that the apoptotic rate of the combination group was significantly higher than that of the other groups, and the cells significantly increased in G1 phase and decreased in S phase (P<0.05). As Western blotting showed, there were no differences in the expressions of AKT and ERK1/2 protein among the four groups (P>0.05); in the LY294002 treatment group, the level of p-AKT protein decreased and p-ERK increased; in the AZD6244 group, the level of p-ERK1/2 protein decreased and p-AKT increased; in the combination group, the levels of p-AKT, p-ERK1/2 and cyclin D1 protein were significantly inhibited, while cleaved caspase-3 protein was up-regulated.. Combined PI3K inhibitor LY294002 and MEK inhibitor AZD6244 has synergistic effect on inhibition of SKOV3/DDP cell growth by inducing apoptosis and blocking cell cycle.

Topics: Antineoplastic Agents; Apoptosis; Benzimidazoles; Blotting, Western; Caspase 3; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Cell Survival; Chromones; Cisplatin; Dose-Response Relationship, Drug; Drug Resistance, Neoplasm; Drug Synergism; Enzyme Inhibitors; Female; Flow Cytometry; Humans; Inhibitory Concentration 50; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinase Kinases; Morpholines; Ovarian Neoplasms; Phosphatidylinositol 3-Kinases; Phosphoinositide-3 Kinase Inhibitors; Proto-Oncogene Proteins c-akt

BRAF and KRAS mutations in ovarian serous borderline tumours (OSBTs) and ovarian low-grade serous carcinomas (LGSCs) have been previously described. However, whether those OSBTs would progress to LGSCs or whether those LGSCs were developed from OSBT precursors in previous studies is unknown. Therefore, we assessed KRAS and BRAF mutations in tumour samples from 23 recurrent LGSC patients with a known initial diagnosis of OSBT. Paraffin blocks from both OSBT and LGSC samples were available for five patients, and either OSBTs or LGSCs were available for another 18 patients. Tumour cells from paraffin-embedded tissues were dissected out for mutation analysis by conventional polymerase chain reaction (PCR) and Sanger sequencing. Tumours that appeared to have wild-type KRAS by conventional PCR-Sanger sequencing were further analysed by full COLD (co-amplification at lower denaturation temperature)-PCR and deep sequencing. Full COLD-PCR was able to enrich the amplification of mutated alleles. Deep sequencing was performed with the Ion Torrent personal genome machine (PGM). By conventional PCR-Sanger sequencing, BRAF mutation was detected only in one patient and KRAS mutations were detected in ten patients. Full COLD-PCR deep sequencing detected low-abundance KRAS mutations in eight additional patients. Three of the five patients with both OSBT and LGSC samples available had the same KRAS mutations detected in both OSBT and LGSC samples. The remaining two patients had only KRAS mutations detected in their LGSC samples. For patients with either OSBT or LGSC samples available, KRAS mutations were detected in seven OSBT samples and six LGSC samples. Surprisingly, patients with the KRAS G12V mutation have shorter survival times. In summary, KRAS mutations are very common in recurrent LGSC, while BRAF mutations are rare. The findings indicate that recurrent LGSC can arise from proliferation of OSBT tumour cells with or without detectable KRAS mutations.

Topics: Adult; Aged; Benzimidazoles; Cell Death; Cystadenocarcinoma, Serous; Cystadenoma, Serous; DNA Mutational Analysis; DNA, Neoplasm; Female; High-Throughput Nucleotide Sequencing; Humans; Kaplan-Meier Estimate; MAP Kinase Kinase Kinases; Middle Aged; Mutation; Neoplasm Grading; Neoplasm Proteins; Neoplasm Recurrence, Local; Ovarian Neoplasms; Prognosis; Protein Kinase Inhibitors; Proto-Oncogene Proteins; Proto-Oncogene Proteins B-raf; Proto-Oncogene Proteins p21(ras); ras Proteins; Tumor Cells, Cultured; Young Adult

Topics: Benzimidazoles; Cystadenocarcinoma, Serous; Female; Humans; Mitogen-Activated Protein Kinase Kinases; Neoplasm Recurrence, Local; Ovarian Neoplasms; Peritoneal Neoplasms

Clear cell carcinoma (CCC) of the ovary tends to show resistance to standard chemotherapy, which results in poor survival for patients with CCC. Developing a novel therapeutic strategy is imperative to improve patient prognosis. Epidermal growth factor receptor (EGFR) is frequently expressed in epithelial ovarian cancer. One of the major downstream targets of the EGFR signaling cascade is extracellular signal-related kinase (ERK). PEA-15, a 15-kDa phosphoprotein, can sequester ERK in the cytoplasm. MEK1/2 plays a central role in integrating mitogenic signals into the ERK pathway. We tested the hypothesis that inhibition of the EGFR-ERK pathway suppresses tumorigenicity in CCC, and we investigated the role of PEA-15 in ERK-targeted therapy in CCC. We screened a panel of 4 CCC cell lines (RMG-I, SMOV-2, OVTOKO, and KOC-7c) and observed that the EGFR tyrosine kinase inhibitor erlotinib inhibited cell proliferation of EGFR-overexpressing CCC cell lines through partial dependence on the MEK/ERK pathway. Furthermore, erlotinib-sensitive cell lines were also sensitive to the MEK inhibitor selumetinib (AZD6244), which is under clinical development. Knockdown of PEA-15 expression resulted in reversal of selumetinib-sensitive cells to resistant cells, implying that PEA-15 contributes to selumetinib sensitivity. Both selumetinib and erlotinib significantly suppressed tumor growth (P < 0.0001) in a CCC xenograft model. However, selumetinib was better tolerated; erlotinib-treated mice exhibited significant toxic effects (marked weight loss and severe skin peeling) at high doses. Our findings indicate that the MEK-ERK pathway is a potential target for EGFR-overexpressing CCC and indicate that selumetinib and erlotinib are worth exploring as therapeutic agents for CCC.

Topics: Adenocarcinoma, Clear Cell; Animals; Apoptosis Regulatory Proteins; Benzimidazoles; Blotting, Western; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Dose-Response Relationship, Drug; ErbB Receptors; Erlotinib Hydrochloride; Female; G1 Phase; Humans; Intracellular Signaling Peptides and Proteins; MAP Kinase Kinase 1; MAP Kinase Kinase 2; MAP Kinase Signaling System; Mice; Mice, Nude; Ovarian Neoplasms; Phosphoproteins; Protein Kinase Inhibitors; Quinazolines; RNA Interference; Tumor Burden; Xenograft Model Antitumor Assays