Outcome In Ovarian Cancer Patients Research Articles

Abstract 4176: The homeobox gene DLX4 stimulates inducible nitric oxide synthase-mediated angiogenesis in ovarian cancer

Abstract Increasing evidence indicates that the free radical gas nitric oxide plays an important role in stimulating angiogenesis. However, the mechanisms that control nitric oxide-mediated angiogenesis in tumors are poorly understood. We previously identified that DLX4, a transcription factor encoded by a homeobox gene, stimulates ovarian tumor progression and correlates with poor outcomes of ovarian cancer patients. In this study, we identified that high DLX4 expression in ovarian cancer specimens is strongly associated with elevated expression of inducible nitric oxide synthase (iNOS), an enzyme that generates nitric oxide. DLX4 induced iNOS expression in ovarian cancer cells in vitro and in xenograft models. We identified that DLX4 interacts with STAT1 and induces iNOS expression in a STAT1-dependent manner. In in vitro studies, inhibition of iNOS abrogated the stimulatory effects of DLX4 on nitric oxide production, vascular endothelial growth factor expression and endothelial cell growth. Furthermore, inhibition of iNOS abrogated the stimulatory effects of DLX4 on tumor microvessel density and ascites formation in ovarian cancer xenograft models. Collectively, our findings indicate that DLX4 contributes to ovarian tumor progression in part by promoting nitric oxide-mediated angiogenesis. This is the first report that functionally links the aberrant expression of a developmental patterning gene in tumors with free radical gas generation. Note: This abstract was not presented at the meeting. Citation Format: Bon Q. Trinh, Song Yi Ko, Dhwani Haria, Nicolas Barengo, Honami Naora. The homeobox gene DLX4 stimulates inducible nitric oxide synthase-mediated angiogenesis in ovarian cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4176. doi:10.1158/1538-7445.AM2015-4176

Abstract 4631: AKAP12 is elevated in paclitaxel-resistant ovarian cancer cells and correlates with poor ovarian cancer patient survival

Abstract Objective: Although most ovarian cancer patients respond initially to the standard of care involving carboplatinum/paclitaxel-based chemotherapy, the majority will recur with chemotherapeutically resistant disease. Unfortunately, no clinical biomarker(s) for predicting or monitoring chemotherapeutic resistance are available. We sought to identify shed and/or secreted proteins from paclitaxel-resistant human ovarian cancer cells and identify whether any of these correlate with ovarian cancer patient outcome. Methods: A proteomic analysis of secretomes from high grade serous ovarian cancer (HGSOC) cells (OV90) was compared to that from a syngeneic paclitaxel-resistant variant (OV90-TR) and a cancer cell line developed from a chemorefractory HGSOC patient. Associations between the identified candidates and patient outcome were assessed from publicly available transcript expression data (n = 545 ovarian cancer patients) and validated independently (n = 795 ovarian cancer patients). Results: Among the 81 differentially abundant proteins identified (q<0.05) from paclitaxel-sensitive vs -resistant HGSOC cell secretomes, AKAP12 was verified to be elevated at the protein and transcript level in all models of paclitaxel-resistant HGSOC. Further, elevated AKAP12 transcript expression was found to significantly correlate with poor progression-free (Hazard Ratio = 1.487 (± 0.148), p = 0.0085 (± 0.002) and overall (Hazard Ratio = 1.22 (± 0.08), p = 0.009 (± 0.013) survival. Conclusions: These findings suggest that elevation and efflux of AKAP12 is characteristic of paclitaxel-resistant HGSOC cells and that elevated AKAP12 expression correlates with poor ovarian cancer patient survival. Citation Format: Nicholas Bateman, Elizabeth Jaworski, Guisong Wang, Elizabeth Dubil, Charlotte Marcus, Kelly Conrads, Pang-ning Teng, Brian Hood, Chad Hamilton, Larry Maxwell, Kathleen Darcy, Thomas Conrads. AKAP12 is elevated in paclitaxel-resistant ovarian cancer cells and correlates with poor ovarian cancer patient survival. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4631. doi:10.1158/1538-7445.AM2015-4631

IL17a and IL21 combined with surgical status predict the outcome of ovarian cancer patients.

Aside from tumor cells, ovarian cancer-related ascites contains the immune components. The aim of this study was to evaluate whether a combination of clinical and immunological parameters can predict survival in patients with ovarian cancer. Ascites specimens and medical records from 144 ovarian cancer patients at our hospital were used as the derivation group to select target clinical and immunological factors to generate a risk-scoring system to predict patient survival. Eighty-two cases from another hospital were used as the validation group to evaluate this system. The surgical status and expression levels of interleukin 17a (IL17a) and IL21 in ascites were selected for the risk-scoring system in the derivation group. The areas under the receiver operating characteristic (AUROC) curves of the overall score for disease-free survival (DFS) of the ovarian cancer patients were 0.84 in the derivation group, 0.85 in the validation group, and 0.84 for all the patients. The AUROC curves of the overall score for overall survival (OS) of cases were 0.78 in the derivation group, 0.76 in the validation group, and 0.76 for all the studied patients. Good correlations between overall risk score and survival of the ovarian cancer patients were demonstrated by sub-grouping all participants into four groups (P for trend <0.001 for DFS and OS). Therefore, acombination of clinical and immunological parameters can provide a practical scoring system to predict the survival of patients with ovarian carcinoma. IL17a and IL21 can potentially be used as prognostic and therapeutic biomarkers.

Venting Gastrostomy at Home for Symptomatic Management of Bowel Obstruction in Advanced/Recurrent Ovarian Malignancy: A Case Series.

Malignant bowel obstruction (MBO) frequently defines the trajectory of end-stage ovarian cancer and results in severe physical and psychological distress in patients and their caregivers. Venting gastrostomy (VG) is an alternative to both prolonged medical therapy with nasogastric intubation and intestinal bypass/diversion for refractory MBO. Limited published data from large academic research centers support use of VG in patients with advanced ovarian cancer and MBO. In this case series, we describe supportive care outcomes in ovarian cancer patients with MBO receiving the effects of VG in a community setting. Six cases of advanced ovarian cancer involving MBO were evaluated for VG from July 2009 through February 2012. Five of six patients were managed with VG. Our experience suggests that VG may be beneficial in controlling nausea and vomiting in ovarian cancer patients with MBO and that VG placement with concurrent evacuation of large-volume ascites was associated with minimal complications. Future prospective studies to evaluate the benefits of VG are warranted.

Eliminating ovarian cancer stem cells: a potential therapeutic target for ovarian cancer chemoresistance.

Tumor recurrence followed by chemoresistance is a major cause of the poor survival rates for ovarian cancer. Despite advances in ovarian cancer chemotherapies and increased knowledge of the characteristics of ovarian cancer, chemoresistance is still an enormous clinical challenge. Current research suggests that ovarian cancer stem cells (OCSCs) might be involved in the occurrence of chemoresistance in ovarian cancer. In this review, we focus on the relationship of ovarian cancer stem cells with drug-resistant ovarian cancer phenotypes. We also discuss the current knowledge regarding ovarian cancer stem cell-based therapeutic strategies as attempts to overcome chemoresistance, the largest obstacle for successful treatment. Finally, we discuss methods for improving the treatment outcomes of ovarian cancer patients with chemoresistance.

Platelet Count After Chemotherapy is a Predictor for Outcome for Ovarian Cancer Patients

Elevated platelet count occasionally is associated with gynecologic malignancies. We investigated the level of platelet count in 450 patients with gynecologic tumors. Ovarian cancer patients have increased platelet count associated with the course of treatment and disease progression. In multivariate analysis, the decrease of platelet count less than 25% after chemotherapy was an unfavorable prognostic factor for PFS (HR, 1.948; 95% CI, 1.083–3.505; p = 0.026) and overall survival (HR, 2.093; 95% CI, 1.022–4.287; p = 0.043). An insufficient decrease of the platelet count increased the risk of recurrence. Thus platelet count could be used for monitoring the disease progression and to predict treatment response.

Abstract A34: MiR-433 induces cellular senescence rendering ovarian cancer cells less likely to undergo chemotherapy-induced apoptosis

Abstract Annually, ovarian cancer (OC) affects 240,000 women worldwide and is the most lethal gynaecological malignancy. Such mortality is predominantly associated with the development of an intrinsic and acquired resistance to chemotherapy, the lack of targeted therapies and the lack of biomarkers predicting therapeutic response. Our clinical data demonstrates that increased miR-433 expression in primary high grade serous OC (HGSOCs) is significantly associated with poor PFS (n=46, p=0.024). Interestingly, the IHC analysis of two miR-433 targets: MAD2 [Furlong et al., 2012 PMID:22069160] and HDAC6 shows that low IHC levels of both proteins is also significantly associated with worse outcome (p=0.002 and 0.002 respectively; n=43). Additionally, the analysis of miR 433 in the publicly available TCGA dataset corroborates that high miR-433 is significantly correlated with worse OS for patients presenting with OC (n=558 and p=0.027). In vitro, in a panel of OC cell lines, higher miR-433 and lower MAD2 and HDAC6 levels were associated with resistance to paclitaxel. To further investigate the role of miR-433 in the cellular response to chemotherapy, we generated an OC cell line stably expressing miR-433, or miR-control. MTT viability assays and Western Blot analyses established that miR-433 cells were more resistant to paclitaxel treatment (50nM) compared to miR-controls. Importantly, we have shown for the first time that miR 433 induced senescence, exemplified by a flattened morphology and down-regulation of phosphorylated Retinoblastoma (p-Rb), a molecular marker of senescence. Surprisingly, miR 433 induced senescence was independent from two well recognised senescent drivers: namely p53/p21 and p16. To explore this further we performed an in silico analysis of seven microRNA platforms which indicated that miR 433 potentially targets Cyclin-dependent kinase CDK6, which promotes sustained phosphorylation of Rb and thus cell cycle progression. In vitro, the overexpression of pre-miR-433 resulted in diminished CDK6 expression demonstrating a novel interaction between miR-433 and CDK6. In conclusion, this study demonstrates that high miR-433 expression predicts poor outcome in OC patients by putatively rendering OC cells resistant to paclitaxel treatment through the induction of cellular senescence identifying this microRNA as a potential marker of chemoresponse. Founding: Health Research Board :HRA- POR/2011/113 Citation Format: Karolina Weiner-Gorzel, Malgorzata Milewska, Danniel Sharpe, Caitlin Beggan, Nazia Faheem, Aloysius McGoldrick, Valerie Toh, Patricia Fitzpatrick, Sharon O'Toole, John O'Leary, Stephen F. Madden, Madeline Murphy, Elaine Kay, Malcolm Kell, Amanda McCann, Fiona Furlong. MiR-433 induces cellular senescence rendering ovarian cancer cells less likely to undergo chemotherapy-induced apoptosis. [abstract]. In: Proceedings of the AACR Precision Medicine Series: Drug Sensitivity and Resistance: Improving Cancer Therapy; Jun 18-21, 2014; Orlando, FL. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(4 Suppl): Abstract nr A34.

Abstract 2864: RPS4X, a new prognostic and predictive biomarker of ovarian and breast cancer

Abstract Epithelial ovarian cancer (EOC) is a disease responsible for more cancer deaths among women in the Western world than all other gynecologic malignancies and breast cancer is leading cause of cancer death among the women worldwide. A major problem with such cancers is chemoresistance; hence the need to identify predictive biomarkers. Interestingly, overexpression of YB-1 in ovarian and breast cancer cells induces cisplatin resistance. Platinum coumpounds like cisplatin is often used in the treatments of these cancers. Y box-binding protein 1 (YB-1) is a multifunctional protein that affects transcription, splicing and translation of mRNA. In this study, we used a tagged YB-1 construct to identify by mass spectrometry the proteins that interacted with YB-1 and required for cisplatin resistance. Using the combination of two bioinformatics databases (Oncomine public microarray and genomic hybridization), we focused on the YB-1 protein partners that are potentially involved in cancer progression. We used the siRNA technique to screen different partners of YB-1 that are directly involve in the cisplatin resistance of MCF7 and MDA-MB-231 cells lines. From this analysis, we found that the RPS4X protein, a new partner of YB-1, would be a good marker of cisplatin resistance in breast cancer patients. Interestingly, the YB-1/RPS4X complex was also found in ovarian cancer cells. Like in the breast cancer cell lines, the depletion of RPS4X protein induced the cisplatin resistance of ovarian cancer cells lines (SKOV3 and OVCAR3). Finally, we used a validate antibodies to assess by immunohistochemistry the protein levels of RPS4X and YB-1 in tumor tissue samples from 192 high-grade serous epithelial ovarian cancer patients. We found that RPS4X correlated significantly with ovarian cancer patient outcome. Taken together, these results suggest that, the YB-1/RPS4X complex is a significant potential target to counteract cisplatin resistance in breast and ovarian cancers. Also, we have established that RPS4X is a new promising prognostic marker for patients with high-grade ovarian cancer. More importantly, RPS4X is shown to be predictive of cisplatin response. Additional immunohistochemistry studies on low-grade ovarian cancers and breast cancers are required to confirm the predictive value of RPS4X. Citation Format: Serges P. Tsofack, Liliane Meunier, Anne-Marie Mes-Masson, Michel Lebel. RPS4X, a new prognostic and predictive biomarker of ovarian and breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2864. doi:10.1158/1538-7445.AM2014-2864

Diagnostic Accuracy of Preoperative Prediction of Malignancy in Ovarian Cysts

In this review, we will critically evaluate the evolution of the diagnostic tools used in the prediction of ovarian malignancy preoperatively, that has shown to get a better outcome for ovarian cancer patients. It will triage women to the best place for their surgery and by ensuring the expertise available to perform the surgery reducing the rate of incomplete primary surgery, that has reflected into improve survival of ovarian cancer patients. The Risk of Malignancy Index (RMI) was the first diagnostic tool developed to help clinicians differentiate between benign and malignant ovarian tumours. Jacob et al. [1] described how it was developed and assesses its performance amongst a cohort 143 patients referred to a London hospital for elective surgical investigation of an adnexal mass. Each patient had five criteria assessed preoperatively: age, menopausal status, and clinical impression score (as determined by an experienced gynaecologist using all of the patient’s preoperative clinical information; out of 5, high = more likely to be malignant), ultrasound score (out of 5, high = more likely to be malignant), and CA-125 level. Following surgery, it was determined that 101 of the patients had benign masses and 42 had malignant tumours. Patients in the malignant group had a higher mean age, were more commonly post-menopausal, and had higher clinical impression scores, ultrasound scores and CA-125 levels than patients in the benign group; all of these differences were statistically significant. However, when used individually, the criteria had inadequate sensitivity and specificity for malignancy. Logistic regression analysis revealed that only menopausal status, ultrasound score and CA-125 levels were significantly and independently related to the likelihood ratio for malignancy. These three criteria were then combined to form the RMI: RMI = U × M × CA-125 U = 0 for ultrasound score of 0 = 1 for ultrasound score of 1 = 3 for ultrasound score of 2-5 M = 1 if premenopausal = 3 if postmenopausal When a cutoff score of 200 was used, the RMI showed excellent diagnostic performance, with a sensitivity of 85% and a specificity of 97%. Patients with an RMI score in excess of 200 had 42 times the background risk of cancer, whilst those with a score less than 200 had 0.15 times the risk. The authors concluded that the RMI was a valuable tool for the preoperative assessment of benign and malignant adnexal masses, and that it gave a higher level of discrimination than could be achieved by individual criteria alone. Overall, this is a good study. The sample size was sufficiently large to allow statistically significant differences between groups to be seen. The criteria used for the RMI were not chosen at

Frequent use of complex surgeries and survival outcomes in ovarian cancer patients: A propensity score analysis from the Korean Gynecologic Oncology Group.

5537 Background: To develop an indicator representing the quality of ovarian cancer surgery and to determine its predictive role in a prognostic model. Methods: From January 2002 to December 2008, the clinical records and computed tomography (CT) data of epithelial ovarian cancer patients were reviewed from the KGOG-3022 dataset. A propensity score was developed from a multivariable logistic model and used as a stratification factor or co-variable in Cox models to balance the differences between the high- and low/intermediate-complexity surgery group. Results: Data for 331 patients from six specialized teaching hospitals were reviewed. Complex surgery, defined as a surgery with a surgical complexity score > 7, was more frequently performed in patients with an ‘upper abdominal extension’ pattern, ‘diffuse peritoneal spread’ pattern, and grade III disease (P < 0.001, 0.017, and 0.001, respectively). To balance the differences, a propensity score for complex surgery was developed. The use of complex surgery ...

Targeted Disruption of the JAK2/STAT3 Pathway in Combination with Systemic Administration of Paclitaxel Inhibits the Priming of Ovarian Cancer Stem Cells Leading to a Reduced Tumor Burden.

Chemotherapy resistance associated with recurrent disease is the major cause of poor survival of ovarian cancer patients. We have recently demonstrated activation of the JAK2/STAT3 pathway and the enhancement of a cancer stem cell (CSC)-like phenotype in ovarian cancer cells treated in vitro with chemotherapeutic agents. To elucidate further these mechanisms in vivo, we used a two-tiered paclitaxel treatment approach in nude mice inoculated with ovarian cancer cells. In the first approach, we demonstrate that a single intraperitoneal administration of paclitaxel in mice 7 days after subcutaneous transplantation of the HEY ovarian cancer cell line resulted in a significant increase in the expression of CA125, Oct4, and CD117 in mice xenografts compared to control mice xenografts which did not receive paclitaxel. In the second approach, mice were administered once weekly with paclitaxel and/or a daily dose of the JAK2-specific inhibitor, CYT387, over 4 weeks. Mice receiving paclitaxel only demonstrated a significant decrease in tumor volume compared to control mice. At the molecular level, mouse tumors remaining after paclitaxel administration showed a significant increase in the expression of Oct4 and CD117 coinciding with a significant activation of the JAK2/STAT3 pathway compared to control tumors. The addition of CYT387 with paclitaxel resulted in the suppression of JAK2/STAT3 activation and abrogation of Oct4 and CD117 expression in mouse xenografts. This coincided with significantly smaller tumors in mice administered CYT387 in addition to paclitaxel, compared to the control group and the group of mice receiving paclitaxel only. These data suggest that the systemic administration of paclitaxel enhances Oct4- and CD117-associated CSC-like marker expression in surviving cancer cells in vivo, which can be suppressed by the addition of the JAK2-specific inhibitor CYT387, leading to a significantly smaller tumor burden. These novel findings have the potential for the development of CSC-targeted therapy to improve the treatment outcomes of ovarian cancer patients.

The Prognostic Value of CXCR4 in Ovarian Cancer: A Meta-Analysis

ObjectiveRecent reports have shown that C-X-C chemokine receptor 4 (CXCR4) is expressed in ovarian cancer and plays an important role in metastasis. However, the prognostic value of CXCR4 in ovarian cancer remains controversial and has not been emphasized. The aim of this study is to evaluate the prognostic significance of CXCR4 in ovarian cancer by performing a meta-analysis.MethodsWe systematically searched for studies evaluating the relationship between CXCR4 expression and the outcome of ovarian cancer patients. Only articles in which CXCR4 expression was detected by immunohistochemical staining were included. Hazard ratios (HRs) and relative risk (RR) with 95% confidence intervals (CIs) were pooled as effect size (ES) across studies for overall survival (OS) and progression-free survival (PFS).ResultsA total of 729 patients from 7 studies (6 articles) were included in this meta-analysis. Our results showed that high CXCR4 expression was significantly associated with poor prognosis in terms of OS (ES, 2.81; 95% CI, 1.16–6.80; p = 0.022) and PFS (ES, 8.48; 95% CI, 2.13–33.70; p = 0.002) in ovarian cancer patients. The association between high CXCR4 expression and poor ovarian cancer prognosis in OS was also statistically significant in subgroups of Asian and III-IV patients constituting 70%.ConclusionsThe present meta-analysis indicated that high CXCR4 expression was associated with poor prognosis in ovarian cancer. More studies, especially larger scale and well-matched researches, are warranted to clarify the prognostic effect of CXCR4 on the outcome of ovarian cancer.

Prognostic value of cancer stem cell marker aldehyde dehydrogenase in ovarian cancer: a meta-analysis.

ObjectiveAldehyde dehydrogenase (ALDH) has recently been reported as a marker of cancer stem-like cells in ovarian cancer. However, the prognostic role of ALDH in ovarian cancer still remains controversial. In this study, we aimed to evaluate the association between the expression of ALDH and the outcome of ovarian cancer patients by performing a meta-analysis.MethodsWe systematically searched for studies investigating the relationships between ALDH expression and outcome of ovarian cancer patients. Only articles in which ALDH expression was detected by immunohistochemical staining were included. A meta-analysis was performed to generate combined hazard ratios (HRs) with 95% confidence intervals (CIs) for overall survival (OS) and disease-free survival (DFS).ResultsA total of 1,258 patients from 7 studies (6 articles) were included in the analysis. Our results showed that high ALDH expression in patients with ovarian cancer was associated with poor prognosis in terms of Os (HR, 1.25; 95% CI, 1.07-1.47; P = 0.005) and DFS (HR, 1.58; 95% CI, 1.00-2.49; P = 0.052), though the difference for DFS was not statistically significant. In addition, there was no evidence of publication bias as suggested by Begg’s and Egger’s tests (Begg’s test, P = 0.707; Egger’s test, P = 0.355).ConclusionThe present meta-analysis indicated that elevated ALDH expression was associated with poor prognosis in patients with ovarian cancer.

Abstract B29: The role of C-Terminal Binding Proteins in BRCA1 silencing and genetic instability of ovarian cancer cells

Abstract Objective: Ovarian cancer is the leading cause of mortality from gynecological malignancy in Western countries. High-grade serous ovarian carcinoma (HGSOC) is characterized by genetic instability and frequently harbors defects in genes such as BRCA1 for homologous recombination (HR) pathway. Our group recently demonstrated that the transcriptional co-repressors, C-Terminal Binding Proteins (CtBP1 and CtBP2), are overexpressed in epithelial ovarian carcinoma and CtBP2 overexpression is associated with poor patient survival. We sought to relate the expression of CtBP to genetic instability and chemoresponse in HGSOC cells. Methods: Gene expression profiling was performed for the control and CtBP2-knockdown ovarian cancer cell lines. Chromatin Immunoprecipitation (ChIP)-qPCR assays and a BRCA1 promoter-luciferase reporter assay were performed to compare the binding of CtBP complex to BRCA1 promoter and the activity of BRCA1 promoter in the ovarian cancer cells. Cell cycle status of the cell lines was determined by flow cytometry. DNA double-strand breaks (DSB) were determined by comet assay. esults: Analysis of gene expression profiles revealed potential functions of CtBP2 in silencing the expression of a panel of genes such as BRCA1 and Fanconi anemia complementation group gene FANCD2 that are involved in HR pathway, cell cycle checkpoints, and genetic instability. ChIP analyses and the luciferase assays strongly suggest that the CtBP2 complex binds to the BRCA1 promoter and represses BRCA1 transcription. Ovarian cancer cell lines with knockdown of either CtBP1 or CtBP2 showed increased expression of BRCA1 protein. CtBP2-knockdown ovarian cancer cells showed increased apoptosis and arrest in cell cycle. These cell lines also showed a significant increase in DSB after gamma irradiation than wild-type control cells in a comet assay. Conclusions: CtBP1 and CtBP2 function in the suppression of a panel of target genes such as BRCA1 and FANCD2 that are involved in cell cycle checkpoints and HR pathway. CtBP-mediated BRCA1 silencing may be a significant contributing mechanism for genetic instability and clinicopathologic development of HGSOC. Targeting this epigenetic mechanism may enhance the sensitivity of HGSOC to DNA damaging agents and contribute to improved outcome of ovarian cancer patients. Citation Format: Shubai Liu, Junzheng Yang, Taymaa May, Yuanyuan Hua, Shi Lu, Kevin Elias, Ronny Drapkin, William Welch, Christopher Crum, Ross S. Berkowitz, Shu-Wing Ng. The role of C-Terminal Binding Proteins in BRCA1 silencing and genetic instability of ovarian cancer cells. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr B29.

COL11A1 promotes tumor progression and predicts poor clinical outcome in ovarian cancer

Biomarkers that predict disease progression might assist the development of better therapeutic strategies for aggressive cancers, such as ovarian cancer. Here, we investigated the role of collagen type XI alpha 1 (COL11A1) in cell invasiveness and tumor formation and the prognostic impact of COL11A1 expression in ovarian cancer. Microarray analysis suggested that COL11A1 is a disease progression-associated gene that is linked to ovarian cancer recurrence and poor survival. Small interference RNA-mediated specific reduction in COL11A1 protein levels suppressed the invasive ability and oncogenic potential of ovarian cancer cells and decreased tumor formation and lung colonization in mouse xenografts. A combination of experimental approaches, including real-time RT-PCR, casein zymography and chromatin immunoprecipitation (ChIP) assays, showed that COL11A1 knockdown attenuated MMP3 expression and suppressed binding of Ets-1 to its putative MMP3 promoter-binding site, suggesting that the Ets-1-MMP3 axis is upregulated by COL11A1. Transforming growth factor (TGF)-beta (TGF-β1) treatment triggers the activation of smad2 signaling cascades, leading to activation of COL11A1 and MMP3. Pharmacological inhibition of MMP3 abrogated the TGF-β1-triggered, COL11A1-dependent cell invasiveness. Furthermore, the NF-YA-binding site on the COL11A1 promoter was identified as the major determinant of TGF-β1-dependent COL11A1 activation. Analysis of 88 ovarian cancer patients indicated that high COL11A1 mRNA levels are associated with advanced disease stage. The 5-year recurrence-free and overall survival rates were significantly lower (P=0.006 and P=0.018, respectively) among patients with high expression levels of tissue COL11A1 mRNA compared with those with low expression. We conclude that COL11A1 may promote tumor aggressiveness via the TGF-β1-MMP3 axis and that COL11A1 expression can predict clinical outcome in ovarian cancer patients.

Inferior Vena Cava Filter Placement and Risk of Hematogenous Distant Metastasis in Ovarian Cancer

Recent studies have suggested that inferior vena cava (IVC) filter placement in cancer patients is associated with decreased survival time after insertion. Causality, however, is yet to be understood. This study evaluates (i) the patterns of recurrence or progression of disease; and (ii) survival outcomes of ovarian cancer patients who underwent IVC filter placement. A total of 274 patients who underwent primary cytoreductive surgery for epithelial ovarian, fallopian tube, and primary peritoneal cancers were identified for analysis. Anatomic location of the first recurrence or progression of disease, progression-free survival, and overall survival were correlated to IVC filter placement status inserted during the perioperative period. Overall, 38 (13.9%) patients underwent perioperative IVC filter insertion, of which 37 (97.4%) were permanently placed. The most common indication was newly diagnosed venous thromboembolism (VTE) (52.6%). Patients with IVC filter placement for VTE were more likely to develop subsequent deep vein thrombosis (25% vs. 7.2%, odds ratio, 4.31, 95% confidence interval, 1.40-13.3, P = 0.019), have hematogenous distant metastasis as the site of first recurrence or progression of disease (12-mo hematogenous distant metastasis ratio, 45.2% vs. 13.6%, hazard ratio, 5.10, 95% confidence interval, 2.35-11.1, P < 0.001, multivariate analysis), and show decreased survival outcomes (median progression-free survival, 5.7 vs. 15.3 mo, P < 0.001: and median overall survival, 22.1 vs. 47.2 mo, P = 0.002, both multivariate analysis) when compared with patients without IVC filter placement. Our results suggested that IVC filter placement for VTE in the perioperative period of primary cytoreductive surgery for ovarian cancer may be associated with increased risk of hematogenous distant metastasis and resulted in decreased survival.

Coordinated Regulation of β-Tubulin Isotypes and Epithelial-to-Mesenchymal Transition Protein ZEB1 in Breast Cancer Cells

The regulation of β-tubulin isotypes, the primary targets for antimitotic chemotherapeutic drugs like taxanes, has implications for drug response and drug resistance. Over the past 15 years, micro-RNAs have been studied widely as regulators of mRNA levels. For example, the tumor suppressor miR-200c was shown in cell culture to target mesenchymal genes, including ZEB1 [ Cochrane ( 2009 ) Mol. Cancer Ther. 8 ( 5 ), 1055 - 1066 ]. In that work, exogenous miR-200c was also shown to reduce β-tubulin class III, one of its predicted targets. Furthermore, decreased miR-200c and increased β-tubulin class III were associated with poor outcomes for ovarian cancer patients [ Cittelly , D.M. ( 2012 ) Mol. Cancer Ther. 11 ( 12 ), 2556 - 2565 ]. Because miR-200c targets the epithelial-to-mesenchymal inducer ZEB1, we wanted to know whether changes in ZEB1 parallel β-tubulin isotype changes, implicating β-tubulin isotypes in ZEB1-associated cell survival pathways. We found coordinated positive feedback regulation of mRNA for ZEB1 and β-tubulin isotype classes I, III, and IVB in MDA-MB-231 breast cancer cells, commonly used as a model for triple-negative breast cancers. Low levels of paclitaxel (40 nM) were found to significantly reduce mRNA levels for these tubulin genes along with a 2-3-fold increase in miR-200c. ZEB1 silencing also reduced β-tubulin isotype classes I, III, and IVB mRNA, whereas upregulation of ZEB1 was associated with increases in these isotype classes. Our work indicates that paclitaxel-induced reduction of ZEB1 and β-tubulin isotypes are, in part, due to increased activity of miR-200c. These results suggest that in aggressive breast cancers, as modeled by MDA-MB-231 cells, β-tubulin class III is a biomarker for cell survival mediated through ZEB1-induced tumor progression pathways.

Self-reinforcing loop of amphiregulin and Y-box binding protein-1 contributes to poor outcomes in ovarian cancer

The Y-box binding protein-1 (YB-1) transcription factor is associated with unfavorable clinical outcomes. However, the mechanisms underlying this association remain to be fully elucidated. We demonstrate that YB-1 phosphorylation, indicative of YB-1 activation, is a powerful marker of outcomes for ovarian cancer patients. In ovarian cancer, YB-1 phosphorylation is induced by activation of the lysophosphatidic acid (LPA) receptor (LPAR) via SRC-dependent transactivation of the epidermal growth factor receptor (EGFR) that is coupled to MAPK/p90 ribosomal S6 kinase (p90RSK), but not phosphatidylinositol 3-kinase (PI3K)/AKT signaling. Activation of the LPAR/SRC/EGFR/MAPK/p90RSK/YB-1 axis leads to production of the EGFR ligand amphiregulin (AREG). AREG induces ongoing YB-1 phosphorylation as well as YB-1-dependent AREG expression, thus constituting an AREG/YB-1 self-reinforcing loop. Disruption of transactivation of the EGFR and the downstream self-reinforcing loop decreases invasiveness of ovarian cancer cells in vitro and limits ovarian cancer growth in xenograft models. These findings established the regulation and significance of YB-1 phosphorylation, therefore further exploration of this signaling axis as a therapeutic avenue in ovarian cancer is warranted.

Abstract 1421: Interleukin-22, a protective factor for ovarian cancer during TNFa-induced apoptosis.

Abstract Ovarian cancer is the 5th leading cause of death from cancer in women. Overall, only 10% of patients with advanced ovarian cancer are curable; despite the advances in chemotherapy the 5-year survival is still less than 30%. Interleukin-22 (IL-22), a member of the IL-10 family of cytokines is part of the innate immune system and is expressed by the Th17 cells. IL-22 has been shown to regulate several processes such as the acute-phase response, activation of the innate immune system, cell migration, differentiation and gene expression. However, its role in cancer immunology, especially in ovarian cancer has not been studied. The aim of this study is to investigate the expression and the role IL-22 in human ovarian cancer. In our work, we analyzed the expression of IL-22 protein in human ovarian cancer tissues and ascites samples using immunohistochemistry and ELISA; we tested the expression of IL-22 receptor on human ovarian cancer cells via Western Blot, and determined the association between disease outcome and IL-22 gene expression (Affymetrix)in patients with human ovarian cancer. We evaluated the effect of IL-22 on cellular proliferation using MTT assay and we assessed the effect of IL-22 on TNF-a induced apoptosis using caspase 3/7 activity (Caspase Glo) assay and 7AAD staining on multiple human ovarian cancer cell lines. Here we demonstrate that IL-22 was expressed in human ovarian cancer tissue (mostly in the stroma) and that the receptor for IL-22 was expressed on human ovarian cancer cells. We found that high expression of IL-22, as well as the IL-22-receptor in the tumor tissue leads to poor disease outcome in ovarian cancer patients. On the contrary, high expression of IL-22-binding protein (antagonist of IL-22) was associated with better survival in ovarian cancer patients. We showed that treatment with human recombinant (hr) IL-22 protein induced the activation of Stat-3, as well as increased the proliferation of human ovarian cancer cell lines. We also found that treatment with hr IL-22 at 100, 50 or even at 10 ng/mL concentrations inhibited TNF-a induced apoptosis in OVCAR-5 ovarian cancer cell line, and these results were reversed by the addition of IL22 binding protein to the culture. We conclude that interleukin-22 is an important factor in the tumor microenvironment of ovarian cancer; it promotes tumor growth by increasing proliferation and serves as a protective factor for ovarian cancer during TNF-s induced apoptosis. These data suggest that IL-22 is a potential therapeutical target and/or biomarker in human ovarian cancer. Citation Format: Klara Balint, Tamara Seedial, Phyllis Gimotty, George Coukos, Andrea Facciabene. Interleukin-22, a protective factor for ovarian cancer during TNFa-induced apoptosis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1421. doi:10.1158/1538-7445.AM2013-1421

Abstract 4843: Genetic variants in Notch signaling pathway and ovarian cancer.

Abstract The Notch signaling pathway drives proliferation, differentiation, apoptosis, cell fate choices, and has a key role in the regulation of stem cells in a variety of cancers. In this study, we applied a pathway-based approach to evaluate the associations between single nucleotide polymorphisms in Notch signaling pathway and ovarian cancer risk, survival, and response to chemotherapy. Towards this, we systematically genotyped 139 SNPs from 10 genes in the pathway in 339 Caucasian ovarian cancer cases and 349 Caucasian healthy controls. Individually, four SNPs were significantly associated with the ovarian cancer risk. The most significant SNP was NOTCH4: rs397081, with an adjusted odds ratio (OR) of 0.38 (95% CI = 0.22-0.66, P=0.00057). A highly significant gene-dosage effect was observed for these top variants (P for trend &amp;lt; 0.00001) with those carrying ≥ 3 risk genotypes having a nearly 6-fold increase in risk (OR = 5.67, 95% CI = 2.58 - 12.44] . Classification and regression tree (CART) analysis further revealed potential higher order gene-gene interactions between these four SNPs, with the highest risk group having almost 7-fold increased risk. Ten SNPs were significantly associated with overall survival, of which the strongest association was identified for NOTCH4: rs3131290 (OR = 0.48, 95% CI = 0.27-0.84). Cumulative effect analysis showed that multiple adverse genotypes had a significant dose-dependent effect on overall survival. The median survival times for individuals carrying ≤3, 4 to 6, and ≥7 of these adverse genotypes were &amp;gt;186.3, 62.82, and 28.58 months, respectively. Similarly, a significant association was identified between a poor response to platinum-based chemotherapy and the number of unfavorable SNPs (P for trend &amp;lt; 0.0001). Our findings suggest that genetic variants in the Notch signaling pathway are associated with ovarian cancer risk and could serve as potential predictors of clinical outcomes in ovarian cancer patients. Citation Format: Enping Xu, Karen H. Lu, Michelle AT Hildebrandt, Maosheng Huang, Xifeng Wu, Dong Liang. Genetic variants in Notch signaling pathway and ovarian cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4843. doi:10.1158/1538-7445.AM2013-4843