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The editors are publishing this note to alert readers to a concern about [this article][1] ([1][2]). Four sets of identical representative images exist: Fig. 4B Hec-1A/Control and Fig. 4B Hec-1A SD+PDGF-AA+HmIgG; Fig. 5B CD31/3G3 and Supplementary Fig. S2 CD31/Paclitaxel; Fig. 5B TUNEL/PBS and

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Mounting clinical and preclinical evidence supports a key role for sustained adrenergic signaling in the tumor microenvironment as a driver of tumor growth and progression. However, the mechanisms by which adrenergic neurotransmitters are delivered to the tumor microenvironment are not well understood. Here we present evidence for a feed-forward loop whereby adrenergic signaling leads to increased tumoral innervation. In response to catecholamines, tumor cells produced brain-derived neurotrophic factor (BDNF) in an ADRB3/cAMP/Epac/JNK-dependent manner. Elevated BDNF levels in the tumor microenvironment increased innervation by signaling through host neurotrophic receptor tyrosine kinase 2 receptors. In patients with cancer, high tumor nerve counts were significantly associated with increased BDNF and norepinephrine levels and decreased overall survival. Collectively, these data describe a novel pathway for tumor innervation, with resultant biological and clinical implications. Significance: Sustained adrenergic signaling promotes tumor growth and metastasis through BDNF-mediated tumoral innervation. Cancer Res; 78(12); 3233–42. ©2018 AACR.

Ovarian transposition has proven to be a safe method for preserving ovarian function in young premenopausal women who require pelvic irradiation for treatment of early stage malignancies. We report 2 cases of ovarian torsion after laparoscopic ovarian transposition in 2 young women scheduled for chemotherapy and radiation therapy for treatment of cervical or vaginal cancer. We believe these are the first such cases reported in the literature. In discussions with patients regarding the risks and potential benefits of ovarian transposition, ovarian torsion should be included as a possible, although rare, complication.

Objective The aim of this study was to evaluate the activity of bevacizumab in a cohort of women with recurrent low-grade serous carcinoma of the ovary or peritoneum. Methods This single-institution retrospective study assessed all patients at MD Anderson Cancer Center with recurrent low-grade serous ovarian or peritoneal cancer who received bevacizumab from 2007 to 2016. Study endpoints included best response, median progression-free survival, median overall survival, and toxicity. Results Forty patients received 45 separate "patient-regimens." Most received bevacizumab in combination with chemotherapy. Complete response (CR) was seen in 7.5%, while 40% had partial responses (PR) and 30% achieved stable disease (SD). Disease progression occurred in nine patients (22.5%). Overall response rate (CR+PR) to bevacizumab-containing regimens was 47.5%. Clinical benefit (CR+PR+SD) was seen in 77.5% of patients. Median progression free survival was 10.2months (95% CI 7.9, 12.4). Median overall survival was 34.6months (95% CI 29.5, 39.7). Fifteen patients discontinued bevacizumab related to toxicity. Conclusions Bevacizumab, most often in combination with chemotherapy, has activity in recurrent low-grade ovarian cancer and should be considered a treatment option for these patients. Further investigation into the most effective chemotherapeutic agent in combination with bevacizumab is warranted.

Although progesterone receptor (PR)–targeted therapies are modestly active in patients with uterine cancer, their underlying molecular mechanisms are not well understood. The clinical use of such therapies is limited because of the lack of biomarkers that predict response to PR agonists (progestins) or PR antagonists (onapristone). Thus, understanding the underlying molecular mechanisms of action will provide an advance in developing novel combination therapies for cancer patients. Nuclear translocation of PR has been reported to be ligand-dependent or -independent. Here, we identified that onapristone, a PR antagonist, inhibited nuclear translocation of ligand-dependent or -independent (EGF) phospho-PR (S294), whereas trametinib inhibited nuclear translocation of EGF-induced phospho-PR (S294). Using orthotopic mouse models of uterine cancer, we demonstrated that the combination of onapristone and trametinib results in superior antitumor effects in uterine cancer models compared with either monotherapy. These synergistic effects are, in part, mediated through inhibiting the nuclear translocation of EGF-induced PR phosphorylation in uterine cancer cells. Targeting MAPK-dependent PR activation with onapristone and trametinib significantly inhibited tumor growth in preclinical uterine cancer models and is worthy of further clinical investigation. Mol Cancer Ther; 17(2); 464–73. ©2017 AACR.

This investigation describes the clinical significance of phosphorylated focal adhesion kinase (FAK) at the major activating tyrosine site (Y397) in epithelial ovarian cancer (EOC) cells and tumor-associated endothelial cells. FAK gene amplification as a mechanism for FAK overexpression and the effects of FAK tyrosine kinase inhibitor VS-6062 on tumor growth, metastasis, and angiogenesis were examined. FAK and phospho-FAKY397 were quantified in tumor (FAK-T; pFAK-T) and tumor-associated endothelial (FAK-endo; pFAK-endo) cell compartments of EOCs using immunostaining and qRT-PCR. Associations between expression levels and clinical variables were evaluated. Data from The Cancer Genome Atlas were used to correlate FAK gene copy number and expression levels in EOC specimens. The in vitro and in vivo effects of VS-6062 were assayed in preclinical models. FAK-T and pFAK-T overexpression was significantly associated with advanced stage disease and increased microvessel density (MVD). High MVD was observed in tumors with elevated endothelial cell FAK (59%) and pFAK (44%). Survival was adversely affected by FAK-T overexpression (3.03 vs 2.06 y, P = 0.004), pFAK-T (2.83 vs 1.78 y, P < 0.001), and pFAK-endo (2.33 vs 2.17 y, P = 0.005). FAK gene copy number was increased in 34% of tumors and correlated with expression levels (P < 0.001). VS-6062 significantly blocked EOC and endothelial cell migration as well as endothelial cell tube formation in vitro. VS-6062 reduced mean tumor weight by 56% (P = 0.005), tumor MVD by 40% (P = 0.0001), and extraovarian metastasis (P < 0.01) in orthotopic EOC mouse models. FAK may be a unique therapeutic target in EOC given the dual anti-angiogenic and anti-metastatic potential of FAK inhibitors.

Bevacizumab is the first antiangiogenic agent to have demonstrated benefit as first-line and maintenance therapy in epithelial ovarian cancer (EOC), with the Gynecologic Oncology Group 218 and ICON 7 phase III trials revealing significantly prolonged progression-free survival (PFS) for carboplatin/paclitaxel plus bevacizumab followed by bevacizumab maintenance versus carboplatin/paclitaxel alone. Results are forthcoming from several phase III maintenance trials of investigational antiangiogenic agents, each evaluating PFS as the primary endpoint: AGO-OVAR12/LUME-Ovar1 (nintedanib [BIBF 1120]), AGO-OVAR16 (pazopanib), and TRINOVA-1, -2, and -3 (AMG 386). Here we review available data and ongoing clinical trials of investigational antiangiogenic agents as maintenance therapy for EOC. Current controversies, including uncertainties regarding the (1) most appropriate clinical trial endpoints, (2) optimal dosing, duration, and timing of therapy (e.g., with first-line chemotherapy and/or as maintenance monotherapy), and (3) feasibility, tolerability, and cost of adding these agents to platinum/taxane regimens are also highlighted.

Trabectedin (ET-743, Yondelis®) is a novel marine antineoplastic alkaloid with a unique mechanism of action. The active substance trabectedin, a tetrahydroisoquinoline alkaloid, is a natural product originally isolated from the Caribbean sea squirt, Ecteinascidia turbinata and is currently manufactured by total synthesis. Trabectedin is licensed by the Spanish pharmaceutical drug company, PharmaMar and co-developed by Johnson & Johnson Pharmaceutical Research and Development, L.L.C., pursuant to a licensing agreement with PharmaMar. Trabectedin is the first anticancer marine-derived drug to be approved by the European Union. In 2007, trabectedin obtained marketing authorization from the European Commission and in many other countries worldwide for the treatment of patients with advanced soft tissue sarcoma (STS) after failure of anthracyclines and ifosfamide, or for those patients who are unsuitable to receive these agents. Based on the recently reported results of a large phase III study (OVA-301) comparing pegylated liposomal doxorubicin (PLD) alone with a combination of PLD and trabectedin in patients with recurrent ovarian cancer, in 2009 the European Commission granted marketing authorization for trabectedin combined with PLD for the treatment of patients with relapsed platinum-sensitive ovarian cancer. The results from OVA-301 showed that the combination of trabectedin and PLD improves progression-free survival and overall response rate over PLD alone with acceptable tolerance in the second-line treatment of recurrent ovarian cancer. In addition, an enhanced activity of trabectedin combined with PLD was observed in platinum sensitive patients, especially in those with a platinum-free interval ranging from 6 to 12 months. Overall, trabectedin-induced toxicities are mainly hematological and hepatic, with grade 3/4 neutropenia and thrombocytopenia observed in approximately 50% and 13% of patients, respectively, and grade 3/4 elevation of liver aminotransferases observed in 40-50% of patients treated with trabectedin. Current efforts are focused on the evaluation of the role of trabectedin in prolonging the platinum-free interval and the identification of predictive factors for patients treated with trabectedin as well as in the development of new trabectedin-based combinations.

Objective Compared with every-3-week paclitaxel (q3T) plus carboplatin, dose-dense weekly paclitaxel (ddT) plus carboplatin improved the survival of ovarian cancer patients. We performed a cost analysis comparing these two regimens. Methods Using a Markov decision model, an acceptable incremental cost-effectiveness ratio (ICER) per progression-free life-year saved (PFLYS) was estimated. Cost of drugs, growth colony-stimulating factors, and transfusions were derived from Medicare reimbursement data. Survival and rates of complications were estimated based on the clinical trial. Results Using a body weight and surface area of an average woman age 63, the estimated cost per cycle of ddT was $107 vs. $80 for q3T. The costs per cycle of combination chemotherapy including treatment administration were $873 for ddT and $535 for q3T. With a median progression-free survival (PFS) of 28months with ddT vs. 17.2months with q3T, the ICER was $5809 per PFLYS for ddT compared with q3T arm. Using a maximum ICER of $100,000 per LYS as cost-effective threshold, the ddT regimen was cost-effective. The ICER was most sensitive to the hazard rate for difference in PFS between the two regimens. A 4-month difference in PFS resulted in a $1200 change of ICER per PFLYS. The ICER was also sensitive to overall survival difference, rate of hematological toxicity, and rate of discontinuation. Conclusions In this economic model, dose-dense weekly paclitaxel is a cost-effective treatment option for advanced ovarian cancer.

To investigate access to surgical care for endometrial cancer in Arizona.The Arizona HealthQuery (AZHQ) data warehouse with claims information on over 7 million patients in Arizona was searched using the International Classification of Disease (ICD-9) codes and Current Procedural Terminology (CPT) codes for endometrial cancer surgery from 2005 to 2008. Coordinates were gathered for patients and hospital to determine the distance traveled, race, insurance and annual caseload per hospital/surgeon were collected. Distance traveled was local (50 miles) or distant (≥ 50 miles) and served as the primary independent variable. Secondary variables included age, race, insurance, surgeon annual volume, and hospital annual volume. Logistic regression for distance traveled was performed for insurance coverage, race, hospital volume, and surgeon volume and expressed as an odds ratio.There were 1532 endometrial cancer surgeries performed at 67 hospitals by 242 surgeons in 15 counties. Most (61%) were performed by high-volume surgeons. Approximately 1 in 5 (19%) of patients traveled greater than 50 miles. Medicare insured patients were twice (OR=2.07, 95% CI=1.38-3.13) and Medicaid patients were three times (OR=3.41, 95% CI=1.89-6.15) as likely to travel over 50 miles. No significant difference was found between uninsured and privately insured patients (OR=0.87, 95% CI=0.45-1.68). Patients were more likely to travel to a high volume facility (OR 2.39, 95% CI=1.26-4.51). Hispanics (OR=2.72, 95% CI=1.72-4.32) and Native Americans (OR=8.60, 95% CI=3.43-21.52) were more likely to travel compared to Caucasians.In Arizona significantly different patterns of care are seen for endometrial cancer surgery based upon insurance coverage, race, surgeon and hospital. Patients travel farther to a high-volume hospital and high-volume surgeon. Hispanics or Native Americans travel farther for care when compared with Caucasians. Patients on government funded insurance plans travel farther for care than patients covered by private insurance or those lacking insurance.

TPS5612Background: Patients with platinum-resistant ovarian cancer, following ≥3 lines of chemotherapy have limited treatment options. NUC-1031, a phosphoramidate transformation of gemcitabine, is ...

Recent studies in patients with ovarian cancer suggest that tumor growth may be accelerated following cessation of antiangiogenesis therapy; however, the underlying mechanisms are not well understood. In this study, we aimed to compare the effects of therapy withdrawal to those of continuous treatment with various antiangiogenic agents. Cessation of therapy with pazopanib, bevacizumab, and the human and murine anti-VEGF antibody B20 was associated with substantial tumor growth in mouse models of ovarian cancer. Increased tumor growth was accompanied by tumor hypoxia, increased tumor angiogenesis, and vascular leakage. Moreover, we found hypoxia-induced ADP production and platelet infiltration into tumors after withdrawal of antiangiogenic therapy, and lowering platelet counts markedly inhibited tumor rebound after withdrawal of antiangiogenic therapy. Focal adhesion kinase (FAK) in platelets regulated their migration into the tumor microenvironment, and FAK-deficient platelets completely prevented the rebound tumor growth. Additionally, combined therapy with a FAK inhibitor and the antiangiogenic agents pazopanib and bevacizumab reduced tumor growth and inhibited negative effects following withdrawal of antiangiogenic therapy. In summary, these results suggest that FAK may be a unique target in situations in which antiangiogenic agents are withdrawn, and dual targeting of FAK and VEGF could have therapeutic implications for ovarian cancer management.

Delta-like ligand 4 (Dll4), one of the Notch ligands, is overexpressed in ovarian cancer, especially in tumors resistant to anti-VEGF therapy. Here, we examined the biologic effects of dual anti-Dll4 and anti-VEGF therapy in ovarian cancer models. Using Dll4-Fc blockade and anti-Dll4 antibodies (murine REGN1035 and human REGN421), we evaluated the biologic effects of Dll4 inhibition combined with aflibercept or chemotherapy in orthotopic mouse models of ovarian cancer. We also examined potential mechanisms by which dual Dll4 and VEGF targeting inhibit tumor growth using immunohistochemical staining for apoptosis and proliferation markers. Reverse-phase protein arrays were used to identify potential downstream targets of Dll4 blockade. Dual targeting of VEGF and Dll4 with murine REGN1035 showed superior antitumor effects in ovarian cancer models compared with either monotherapy. In the A2780 model, REGN1035 (targets murine Dll4) or REGN421 (targets human Dll4) reduced tumor weights by 62% and 82%, respectively; aflibercept alone reduced tumor weights by 90%. Greater therapeutic effects were observed for Dll4 blockade (REGN1035) combined with either aflibercept or docetaxel (P < 0.05 for the combination vs. aflibercept). The superior antitumor effects of REGN1035 and aflibercept were related to increased apoptosis in tumor cells compared with the monotherapy. We also found that GATA3 expression was significantly increased in tumor stroma from the mice treated with REGN1035 combined with docetaxel or aflibercept, suggesting an indirect effect of these combination treatments on the tumor stroma. These findings identify that dual targeting of Dll4 and VEGF is an attractive therapeutic approach. Mol Cancer Ther; 15(6); 1344–52. ©2016 AACR.

Purpose: We hypothesized that platelet levels during therapy could serve as a biomarker for response to therapy and that manipulation of platelet levels could impact responsiveness to chemotherapy. Experimental Design: The medical records of patients with recurrent or progressive ovarian cancer were retrospectively queried for changes in platelet and CA-125 levels during primary therapy. In vitro coculture experiments and in vivo orthotopic models of human ovarian cancer in mice were used to test the effect of modulating platelet levels on tumor growth and responsiveness to docetaxel. Results: Thrombocytosis at the diagnosis of ovarian cancer was correlated with decreased interval to progression (P = 0.05) and median overall survival (P = 0.007). Mean platelet levels corrected during primary therapy and rose at recurrence. Contrary to treatment-responsive patients, in a cohort of patients refractory to primary therapy, platelet levels did not normalize during therapy. In A2780, HeyA8, and SKOV3-ip1 ovarian cancer cell lines, platelet coculture protected against apoptosis (P < 0.05). In orthotopic models of human ovarian cancer, platelet depletion resulted in 70% reduced mean tumor weight (P < 0.05). Compared with mice treated with docetaxel, mice treated with both docetaxel and platelet-depleting antibody had a 62% decrease in mean tumor weight (P = 0.04). Platelet transfusion increased mean aggregate tumor weight 2.4-fold (P < 0.05), blocked the effect of docetaxel on tumor growth (P = 0.55) and decreased tumor cell apoptosis. Pretransfusion aspirinization of the platelets blocked the growth-promoting effects of transfusion. Conclusions: Platelet-driven effects of chemotherapy response may explain clinical observations. Clin Cancer Res; 21(3); 602–10. ©2014 AACR.

Background To determine the protein expression profile (PEP) of primary and recurrent ovarian cancer patients in order to predict therapeutic targets for chemotherapy. Methods Tissue samples were submitted for PEP in two formats, including formalin-fixed paraffin-embedded tissue for immunohistochemistry (IHC) and fresh frozen tissue for oligonucleotide microarray (MA) gene expression assays. Specimens were analyzed for 18 protein markers and 88 MA genes. A series of Generalized Linear Models (GLM) was used to predict the proportion of positive results by histology for each biomarker. Results Four hundred and twenty-eight specimens were analyzed for IHC and 67 specimens for MA analysis. The majority of specimens, 82%, were serous histology and 35.3% of specimens were poorly differentiated. Sixty percent of specimens were advanced stage, 62% were from a primary diagnosis, and 53% were obtained from a metastatic site. BCRP, ER, MGMT, and RRM1 proteins were overexpressed in 85%, 47%, 93%, and 47% of serous carcinomas, respectively. The MGMT and RRM1 biomarkers were significantly overexpressed in serous (p

There is growing recognition of the important role of metronomic chemotherapy in cancer treatment. On the basis of their unique antiangiogenic effects, we tested the efficacy of nab-paclitaxel, which stimulates thrombospondin-1, and topotecan, which inhibits hypoxia-inducible factor 1-α, at metronomic dosing for the treatment of ovarian carcinoma. In vitro and in vivo SKOV3ip1, HeyA8, and HeyA8-MDR (taxane-resistant) orthotopic models were used to examine the effects of metronomic nab-paclitaxel and metronomic topotecan. We examined cell proliferation (Ki-67), apoptosis (cleaved caspase-3), and angiogenesis (microvessel density, MVD) in tumors obtained at necropsy. In vivo therapy experiments demonstrated treatment with metronomic nab-paclitaxel alone and in combination with metronomic topotecan resulted in significant reductions in tumor weight (62% in the SKOV3ip1 model, P < 0.01 and 96% in the HeyA8 model, P < 0.03) compared with vehicle (P < 0.01). In the HeyA8-MDR model, metronomic monotherapy with either cytotoxic agent had modest effects on tumor growth, but combination therapy decreased tumor burden by 61% compared with vehicle (P < 0.03). The greatest reduction in MVD (P < 0.05) and proliferation was seen in combination metronomic therapy groups. Combination metronomic therapy resulted in prolonged overall survival in vivo compared with other groups (P < 0.001). Tube formation was significantly inhibited in RF-24 endothelial cells exposed to media conditioned with metronomic nab-paclitaxel alone and media conditioned with combination metronomic nab-paclitaxel and metronomic topotecan. The combination of metronomic nab-paclitaxel and metronomic topotecan offers a novel, highly effective therapeutic approach for ovarian carcinoma that merits further clinical development. Mol Cancer Ther; 14(12); 2677–86. ©2015 AACR.

Purpose: Residual disease following primary cytoreduction is associated with adverse overall survival in patients with epithelial ovarian cancer. Accurate identification of patients at high risk of residual disease has been elusive, lacking external validity and prompting many to undergo unnecessary surgical exploration. Our goal was to identify and validate molecular markers associated with high rates of residual disease. Methods: We interrogated two publicly available datasets from chemonaïve primary high-grade serous ovarian tumors for genes overexpressed in patients with residual disease and significant at a 10% false discovery rate (FDR) in both datasets. We selected genes with wide dynamic range for validation in an independent cohort using quantitative RT-PCR to assay gene expression, followed by blinded prediction of a patient subset at high risk for residual disease. Predictive success was evaluated using a one-sided Fisher exact test. Results: Forty-seven probe sets met the 10% FDR criterion in both datasets. These included FABP4 and ADH1B, which tracked tightly, showed dynamic ranges >16-fold and had high expression levels associated with increased incidence of residual disease. In the validation cohort (n = 139), FABP4 and ADH1B were again highly correlated. Using the top quartile of FABP4 PCR values as a prespecified threshold, we found 30 of 35 cases of residual disease in the predicted high-risk group (positive predictive value = 86%) and 54 of 104 among the remaining patients (P = 0.0002; OR, 5.5). Conclusion: High FABP4 and ADH1B expression is associated with significantly higher risk of residual disease in high-grade serous ovarian cancer. Patients with high tumoral levels of these genes may be candidates for neoadjuvant chemotherapy. Clin Cancer Res; 20(12); 3280–8. ©2014 AACR.

Background While most gynecologic cancers respond to first-line cytotoxic chemotherapy, treatment of recurrent disease is frequently associated with acquired drug resistance. In order to find an in vitro surrogate of this clinical phenomenon, a tumor chemoresponse assay was studied. Methods/Materials Patients who had tissue submitted for repeated chemoresponse testing were identified through a retrospective search. Sixty-three patients met inclusion criteria (chemoresponse testing completed at primary diagnosis and upon recurrence of disease and assays completed ≥90 days apart). The Wilcoxon signed-rank test was used to compare chemoresponse, represented as a response index (RI), between primary and recurrent measurements. In a secondary analysis, response was categorized and coded as Responsive = 3, Intermediately Responsive = 2 and Non-Responsive = 1, and the paired t-test was used to compare chemoresponse between primary and recurrent measurement. Results Median time between primary and recurrent tumor testing was 309 days (IQR 208–422). Drugs tested included carboplatin, cisplatin, docetaxel, doxorubicin, gemcitabine, paclitaxel, topotecan, and combination carboplatin/gemcitabine and carboplatin/paclitaxel. There were no differences in chemoresponse between primary and recurrent measurement when chemoresponse was represented by RI scores; although a trend toward increased resistance to paclitaxel upon recurrence was noted. When chemoresponse was analyzed as a continuous variable corresponding to categorized response, a significant shift toward increased resistance to paclitaxel at recurrence, and a marginally significant trend toward increased resistance to carboplatin at recurrence, were observed. Conclusions We observed a trend toward increased chemoresistance at recurrence for paclitaxel, and a marginally significant trend toward increased chemoresistance to carboplatin, but no change in chemoresponsiveness between primary diagnosis and recurrence of disease for other common chemotherapy drugs, including common second-line agents such as doxorubicin, gemcitabine, and topotecan.

Purpose:The purpose of this study was to identify molecular predictors of residual disease (RD) in high grade serous ovarian cancer (HGSC) and further understand their role in promoting cancer metastasis. Method:The current study analyzed Affymetrix gene expression data of 504 HGSC cases from The Cancer Genome Atlas (TCGA) data to identify differentially expressed genes in tumors from patients with no gross residual disease after surgery (NRD) or presence of RD following initial debulking surgery. It was followed by qRT-PCR analysis of tumor samples for validation purposes. RPPA data of 354 patients from TCGA were analyzed. Immunohistochemical analysis was performed on the patient samples to determine the expression at the protein level (cancer versus stromal cells). Gene array was carried out after overexpressing the selected gene in ovarian cancer cells and the data was analyzed by Ingenuity Pathway Analysis (IPA). In vitro (migration and invasion) and in vivo (orthotopic mouse models) assays were used to determine the biological roles of gene(s) identified from the above analyses. Results: In TCGA data set, 97/107 (90.6%) of the patients with high expression of FABP4 gene had residual disease. In the validation cohort, among the 35 patients predicted to be at high risk for residual disease, 30 (86%) did have residual disease. In contrast, only 54 of the 104 patients with FABP4 values below the decision threshold (52%) had incomplete resection (p = 0.0002). RPPA analysis indicated that expression of FABP4 was positively correlated (Spearman correlation analysis) with expression of several other proteins known to increase tumor cell infiltration and metastasis such as JNK2 (p = 0.194), transglutaminase (p = 0.199), c-kit (p = 0.173), fibronectin (p = 0.364), PKC-A (p = 0.178), collagen-6 (p = 0.197) and paxillin (p = 0.239). It was negatively correlated with E-cadherin (p = -0.246) and claudin-7 (p = -0.201) expression. Immunohistochemical analysis confirmed that apart from endothelial cells and adipocytes, cancer cells also express significant amount of FABP4. In vitro assays showed significant reduction in invasion and migration after silencing FABP4 in HGSC cell lines (p Conclusion:These findings provide a new understanding of ovarian cancer metastasis and identify a potentially important target for therapeutic intervention. Citation Format: Kshipra M. Gharpure, Susan L. Tucker, Shelley M. Herbrich, Anna K. Unruh, Alpa M. Nick, Erin K. Crane, Robert L. Coleman, Jamie Guenthoer, Heather J. Dalton, Sherry Y. Wu, Rajesha Rupaimoole, Gabriel Lopez-Berestein, Bulent Ozpolat, Cristina Ivan, Wei Hu, Keith Baggerly, Anil Sood. Mechanistic and functional implications of FABP4 in ovarian cancer metastasis. [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 2273. doi:10.1158/1538-7445.AM2015-2273

Increased adrenergic signaling is known to promote tumor progression, but the underlying mechanisms remain poorly understood. Tumor associated macrophages (TAMs) are key components of the tumor microenvironment that contribute to pro-inflammatory processes and tumor growth. Recently, it has been reported that patients with higher levels of adrenergic signaling have higher counts of MMP-9-producing TAMs. Here, we examined factors responsible for stress-mediated effects on monocyte/macrophage recruitment into the tumor microenvironment and the resultant effects on tumor growth. Conditioned media from norepinephrine- or epinephrine-treated ADRB positive SKOV3ip1 ovarian cancer cells had significantly increased levels of pro-inflammatory cytokines, such as MCP1, fractalkine, IL6, IL8 and VEGF. MCP1, a key modulator of monocyte/macrophage recruitment, gene and protein levels were significantly increased in vitro after catecholamine exposure, which was mediated by cAMP and PKA, while this effect was abrogated by a beta-blocker. SKOV3ip1 tumor samples from mice, subjected to daily restraint stress, had elevated MCP1 gene and protein expression levels, increased CD14+ cells, and increased infiltration of CD68+ cells. Both, propranolol, a non-specific beta-blocker, and hMCP1 siRNA packaged in DOPC nanoparticles significantly abrogated daily restraint stress-induced tumor growth and inhibited infiltration of CD68+ and F4/80+ cells. Of patient samples evaluated (n = 462), 28% had elevated absolute peripheral blood monocyte (>0.7 × 106/μL) and 43% had high peripheral blood monocytes when measured as the percent of white blood cell content (>8%). Elevated peripheral blood monocytes and increased CD68+ cells (>15/hpf) in samples from ovarian cancer patients were significantly associated with advanced disease and worse overall survival. In summary, increased adrenergic signaling is associated with enhanced macrophage infiltration mediated by tumor cell-derived MCP1 production. Citation Format: Guillermo N. Armaiz-Pena, Vianey Gonzalez-Villasana, Archana S. Nagaraja, Cristian Rodriguez-Aguayo, Piotr Dorniak, Rebecca Previs, Nouara Sadaoui, Rebecca Stone, Koji Matsuo, Heather J. Dalton, Susan K. Lutgendorf, Anil K. Sood, Gabriel Lopez-Berestein. Adrenergic regulation of MCP-1 leads to enhanced macrophage recruitment and ovarian carcinoma growth. [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 2359. doi:10.1158/1538-7445.AM2015-2359

Purpose: To determine the effects of Dll4 blockade with and without aflibercept on ovarian tumor burden and ascites in preclinical models. Methods: Using Dll4-Fc and anti-Dll4 antibodies, we evaluated the biological effects of Dll4 inhibition combined with aflibercept in vivo (orthotopic mouse models of ovarian cancer). Reverse phase protein array (RPPA) analysis was used to identify potential biomarkers of treatment response. Results: To address the biological significance of Dll4 expression in tumor cells versus endothelial cells, we used human and mouse anti-Dll4 antibodies in vivo. In the A2780 model, treatment with mouse (REGN1035) or human (REGN421) anti-Dll4 antibodies reduced tumor weight by 61.5% and 82.4%, respectively, compared to the controls (P < 0.05 for both). Aflibercept alone reduced tumor weight by 90% (p < 0.05). The combinations of REGN1035 plus aflibercept, and REGN421 plus aflibercept resulted in the greatest inhibition of tumor growth, reducing respective tumor weight by 95.8% and 93.9%, compared to the controls (P < 0.05). The anti-tumor effects were related to decreased angiogenesis and increased apoptosis. RPPA analysis revealed that caspase3 and GATA3 were significantly increased; cyclin D1, Bcl-2 and pS6-s240_s244 were decreased in response to Dll4 Inhibition plus aflibercept. Among these, GATA3, a transcription factor for E-cadherin, was significantly induced by Dll4 Inhibition plus aflibercept, resulting in the up-regulation of E-cadherin expression in human cancer cells. Furthermore, GATA3 and E-cadherin expression might be useful as potential biomarkers of response. Conclusions: Dual targeting of Dll4 and VEGF shows promise for inhibiting ovarian tumor growth and may have important clinical implications. Citation Format: Jie Huang, Wei Hu, Heather J. Dalton, Justin Bottsford-Miller, Robert L. Coleman, Robert B. Jaffe, Anil K. Sood. Dll4 inhibition plus aflibercept markedly reduces ovarian tumor burden and ascites. [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 2993. doi:10.1158/1538-7445.AM2014-2993

Objectives: Bisphosphonates, known to have beneficial effects on bone and soft tissue metastases in breast cancer, are potent inhibitors of macrophages, but effects on the microenvironment are not fully understood. We investigated the biological effect of clodronate on macrophage and endothelial cell-driven angiogenesis in ovarian cancer. Methods: Using the FDA Adverse Event Reporting System (FAERS), we examined the effects of bisphosphonate use on overall cancer mortality. We examined the in vitro (endothelial cell migration, capillary formation and cytokine secretion) and in vivo (orthotopic mouse models) effects of clodronate on angiogenesis, macrophage infiltration, and tumor growth. Results: Using (FAERS) data, out of ∼17,000 patients with a cancer diagnosis co-medicated with a bisphosphonate, overall reported death rate was 36% lower (17.6% vs 27.7%, p Conclusions: Bisphosphonates modulate tumor angiogenesis through effects on macrophages and endothelial cells and are associated with overall decreased mortality in cancer patients, independent of chemotherapeutic agents. Bisphosphonates represent an unexplored, but attractive clinical strategy in ovarian cancer, with potential for synergistic combination with other anti-angiogenic agents. Citation Format: Heather J. Dalton, Nicole M. Reusser, Alexander Zien, David Jackson, Rebecca Previs, Rajesha Rupaimoole, Behrouz Zand, Gabriel Lopez-Berestein, Robert L. Coleman, Anil K. Sood. Bisphosphonates: New strategies for targeting angiogenesis in ovarian 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 2983. doi:10.1158/1538-7445.AM2014-2983

Purpose: Paxillin (PXN) expression is increased in various cancer types, but its biological and clinical impact is not well understood. Here, we examined the biological effects of paxillin in ovarian cancer. Methods: The in vivo (HeyA8 and HeyA8-MDR orthotopic mouse models of ovarian cancer) and in vitro (tube formation assays, reverse phase protein array (RPPA) analysis, RT-PCR chromatin-immunoprecipitation (ChIP) assay) effects of paxillin were examined in ovarian cancer. Results: Paxillin silencing using siRNA incorporated in DOPC nanoliposomes resulted in 55% reduction in tumor growth in a HeyA8 orthotopic ovarian cancer mouse model (p Conclusion PXN exerts indirect effects on tumor angiogenesis, mediated through Src. These findings provide a new understanding of the role of PXN in malignant biology. Citation Format: Hyun Jin Choi, Seung-Wook Kim, Sunila Pradeep, Heather J. Dalton, Rajesha Rupaimoole, Selanere Mangala, Gabriel Lopez, Sood K. Anil. Paxillin enhances angiogenesis through transcriptional regulation of Src in ovarian 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 2. doi:10.1158/1538-7445.AM2014-2

Purpose: The purpose of this study was to investigate the combined antitumor effects of metronomic docetaxel-containing PLGA-PRINT nanoparticles and antiangiogenic mEZH2 siRNA incorporated into chitosan nanoparticles (CH-mEZH2 siRNA). Method: In vivo dose-finding studies and therapeutic experiments with PLGA-PRINT-docetaxel and CH-mEZH2 siRNA were conducted in well-established orthotopic mouse models of ovarian cancer (HeyA8 and SKOV3ip1). Antitumor effects were quantified through mean tumor weight and tumor nodule counts. Tumor tissues from these studies were stained to evaluate the proliferation index (Ki67), apoptosis index (cleaved caspase-3), and microvessel density (CD31). Result: Dose-finding studies for PLGA-PRINT-docetaxel revealed all doses (0.5, 1.5, 2 mg/kg for metronomic and 20mg/kg for MTD) significantly reduced tumor burden (p Citation Format: Kshipra M. Gharpure, Kevin S. Chu, Charles Bowerman, Takahito Miyake, Sunila Pradeep, Lingegowda S. Mangala, Hee-Dong Han, Rajesha Rupaimoole, Sherry Y. Wu, Heather J. Dalton, Mary E. Napier, Gabriel Lopez-Berestein, Joseph M. DeSimone, Anil K. Sood. Metronomic docetaxel in PRINT nanoparticles and EZH2 silencing have synergistic antitumor effect in ovarian 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 5403. doi:10.1158/1538-7445.AM2014-5403

Objective: Chronic stress and sustained adrenergic signaling promote tumor progression. The underlying mechanisms behind this process are not well understood. We examined the effects of sustained adrenergic signaling on cervical cancer progression through increased expression of HPV oncogenes, E6 and E7. Methods: Beta-adrenergic receptor (ADRB) positive cervical cancer cell lines (CaSki and SiHa) were treated with norepinephrine (NE) or isoproterenol (ISO) to analyze intracellular responses. Migration, invasion and anoikis assays were performed to elucidate the resultant biological effect of NE and ISO. ADRB expression levels were examined from cervical cancer patient tumor samples. Results: ADRB expression was determined for cervical cell lines (CaSki, SiHa, and C33A) using qRT-PCR. CaSki and SiHa cells express ADRB1, ADRB2, and ABRB3. Cells exposed to NE for 30 min showed elevated cyclic AMP activity. After treatment with NE or ISO, mRNA levels from HPV oncogenes, E6 and E7, were significantly elevated in the SiHa and CaSki cells. NE exposure resulted in a significant increase in invasion and migration of cervical cancer cells, while E6 siRNA abrogated these effects. After 48 hours of NE or ISO exposure, CaSki cells showed a 30% reduction in anoikis. Among 166 tumor samples evaluated from cervical cancer patients, 85% had increased ADRB1 expression and 61% had increased ADRB2 expression. Tumor stage or grade was not related to ADRB expression. ADRB1 expression was not correlated with patient survival outcomes (p = 0.86); however, patients with high ADRB2 expressed had decreased overall survival (p=0.038). Conclusion: Increased adrenergic signaling promotes cervical cancer progression. Disruption of this pathway could provide a novel complement to current therapies. Citation Format: Nouara C. Sadaoui, Guillermo N. Armaiz-Pena, Archana S. Nagaraja, Rajesha Rupaimoole, Rebecca A. Previs, Heather J. Dalton, Mangala S. Lingegowda, Lois M. Ramondetta, Anil K. Sood, Susan K. Lutgendorf, Steve W. Cole. Sustained adrenergic signaling promotes cervical cancer progression. [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 3511. doi:10.1158/1538-7445.AM2014-3511

Targeted anti-angiogenic therapies were a highly anticipated addition to cancer treatment, but their clinical use has been tempered by the development of resistance. To understand the role of immune cells in potentially mediating such resistance, we carried out systematic immune cell profiling in a syngeneic ovarian cancer mouse model following treatment with the B20 anti-VEGF antibody. Following injection of luciferase-labeled cancer cells and initiation of treatment, mice (n=8/group) were divided into B20-sensitive and resistant groups based on longitudinal bioluminescence imaging of tumor burden. Immune cell profiling confirmed a dramatic increase in macrophages with emergence of resistance to B20. Using the syngeneic model, resistance to B20 was documented with bioluminescence imaging as an increase in tumor burden in mice with previously stable disease. Zoledronic acid, a macrophage-depleting bisphosphonate, was then added to B20 treatment. Restoration of sensitivity to B20 with the addition of zoledronic acid was demonstrated by a 68% prolongation of survival in this treatment group as compared to those receiving B20 alone (p Citation Format: Heather J. Dalton, Sunila Pradeep, Guillermo N. Armaiz-Pena, Rebecca Previs, Ashley Davis, Rajesha Rupaimoole, Behrouz Zand, Yared Hailemichael, Willem W. Overwijk, Gabriel Lopez-Berestein, Anil K. Sood. Macrophages modulate adaptive resistance to anti-angiogenic therapy. [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 2982. doi:10.1158/1538-7445.AM2014-2982

Objective: The Notch pathway plays a critical role in cell-to-cell communication, proliferative signaling, and cell differentiation, but can have oncogenic or tumor suppressive effects depending on the tissue type. Dysregulation of the PI3K/AKT pathway has been implicated in a variety of cancers and represents an attractive therapeutic target. We analyzed the clinical significance and interplay of these two pathways in endometrial cancer. Methods: We accessed clinical, somatic mutations, and Reverse Phase Protein Array (RPPA) data from The Cancer Genome Atlas (TCGA) to perform integrated analyses and determine the clinical significance of Notch pathway aberrations in uterine cancer patients. In this cohort, we also identified concomitant PI3K/AKT abnormalities. Clinical information extracted included age, BMI, tumor histology, tumor grade, clinical stage, estrogen(ER)/progesterone(PR) receptor status, and overall survival. Results: A total of 232 samples were available for analysis. Within the Notch pathway, 18.1% of evaluable samples had amplification or upregulation of Notch2/Notch3 and/or DLL3 genes, which was significantly correlated with worse overall survival (p=3.08e-7). Median ERα, ERα(pS118), and PR RPPA levels were significantly higher for patients with no abnormalities or mutations present in Notch2/Notch 3 and/or DLL3 than those with amplification and/or upregulation of these genes (p=0.0016, p=0.0002, and p=0.003, respectively). Of the 52 identified mutations in Notch2/Notch3 and/or DLL3, 92.3% and 7.7% were missense and nonsense mutations, respectively. PI3KCA represented the most frequently mutated gene within this subset affecting 59% of patients; 159 mutations were identified and 95% were missense mutations. Overexpression or amplification of mRNA involved in the PI3K/AKT pathway was present in 108 (46.5%) of the evaluated endometrial tumors. Of these, only 35 tumors (32.4%) had overlapping, concomitant Notch2/Notch3/DLL3 mRNA overexpression or amplification. Conclusions: Upregulation or amplification of Notch 2, Notch 3, and/or DLL3 genes predicted a more aggressive clinical course for endometrial cancer patients. Given the non-overlapping expression of PI3K/AKT and Notch pathways, personalized options for targeted therapy could be considered for each group. Citation Format: Rebecca A. Previs, Cristina Ivan, Heather J. Dalton, Ashley N. Davis, Justin N. Bottsford-Miller, Behrouz Zand, Michael H. McGuire, Guillermo N. Armaiz-Pena, Robert L. Coleman, Keith A. Baggerly, Anil K. Sood. Exploiting the non-overlapping dysregulation of Notch and PI3K/AKT signaling pathways as a guide for personalizing uterine cancer therapy. [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 3340. doi:10.1158/1538-7445.AM2014-3340

Mounting clinical and preclinical evidence supports a key role of sustained adrenergic signaling in the tumor microenvironment as a driver of tumor growth and progression. However, the mechanisms by which adrenergic neurotransmitters are delivered to the tumor microenvironment are not well understood. Sustained adrenergic signaling resulted in increased tumor growth and was correlated with a significant increase in intratumoral nerve density and norepinephrine in several orthotopic models of disease. These effects were completely abrogated by hexamethonium bromide (ganglionic blocker), but not by removal of the adrenal glands. To elucidate mechanism by which tumor cells induce nerve growth, we analyzed gene expression changes and found BDNF to be a key factor upregulated by catecholamine treatment. Treatment with different ADRB agonist and blockers determined that ADRB3 is required for the induction of BDNF. Use of cAMP and Epac agonist/antagonist demonstrated that this axis is required for catecholamine induced BDNF expression. Further analyses revealed that JNK exerts its control of BDNF expression through the regulation of AP-1. In various orthotopic models of disease, chronic stress significantly increased tumor growth and intratumoral innervation, an effect completely blocked by BDNF siRNA-DOPC nanoliposomes. Moreover, BNDF overexpression was sufficient to induce tumor growth and intratumoral innervation. Silencing of TrkB, BDNF receptor, on the host by murine BDNF siRNA-Chitosan nanoparticles blocked the effects of sustained adrenergic signaling on tumor growth and innervation. Additionally, inhibition of host TrkB activity in a transgenic TrkB Kinase-Switch mouse model inoculated with ovarian cancer cells showed that adrenergic induced tumor growth and innervation is mediated by the host TrkB. In cancer patients, high tumor nerve density was significantly associated with increased BDNF and norepinephrine levels, and worse overall survival. Collectively, these data describe a novel pathway for tumor neo-innervation with resultant biological and clinical implications. Citation Format: Guillermo N. Armaiz-Pena, Archana S. Nagaraja, Julie K. Allen, Nouara C. Sadaoui, Rajesha Rupaimoole, sherry Y. wu, Sunila Pradeep, Hee Dong Han, Behrouz Zand, Heather Dalton, Rebecca Previs, Morgan Taylor, Justin Bottsford-Miller, Lingegowda S. Mangala, Cristian Rodriguez-Aguayo, Mariela De Biasi, Gabriel Lopez-Berestein, Steve Cole, Susan K. Lutgendorf, Anil K. Sood. Sustained adrenergic signaling promotes intratumoral innervation through BDNF induction. [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 1062. doi:10.1158/1538-7445.AM2014-1062

Objective: There is growing recognition of the important role of metronomic chemotherapy for cancer treatment. Based on their unique anti-angiogenic effects, we tested the efficacy of dual albumin-bound paclitaxel (ABP), which stimulates Thrombospondin-1, and topotecan, which inhibits Hypoxia-inducible factor 1-α, chemotherapy at metronomic dosing in ovarian carcinoma. Methods: We examined the effects of ABP and topotecan in vitro (MTT) and in vivo using SKOV3ip1, HeyA8 and HeyA8-MDR orthotopic mouse models of ovarian cancer. We also examined effects on proliferation (Ki67), apoptosis (TUNEL), and angiogenesis (MVD) in tumor samples obtained at necropsy. Results: In vitro cytotoxicity assays revealed similar effects (IC50=20-60 nM) of maximum tolerated dosing (MTD) and metronomic dosing of ABP on HeyA8 tumor cells (p Conclusions: Metronomically-dosed albumin-bound paclitaxel and topotecan offer a highly effective and novel therapeutic approach in ovarian carcinoma that merits further clinical development. Citation Format: Ashley N. Davis, Yvonne G. Lin-Liu, Rebecca A. Previs, Heather J. Dalton, Behrouz Zand, Justin Bottsford-Miller, Robert Coleman, Anil K. Sood. Metronomic albumin-bound paclitaxel and topotecan has potent antitumor activity in ovarian 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 2995. doi:10.1158/1538-7445.AM2014-2995

Objectives: Gross residual disease (RD) following primary cytoreduction is the best predictor of overall survival in patients with high-grade serous ovarian cancer (HGSOC). Accurate identification of patients who will have RD has been elusive, prompting many to undergo unnecessary surgical exploration. Our goal was to identify and validate molecular markers associated with high rates of RD. Methods: We interrogated two publicly available genomic datasets of primary HGSOC for genes consistently differentially expressed in RD and no residual disease (R0) cohorts and significant at a false-discovery rate (FDR) of 10%. Genomic expression was further validated in an independent cohort (chemonaive ovarian tumor tissues) using quantitative reverse-transcriptase polymerase chain reaction followed by a blinded prediction of RD. A one-sided Fisher’s exact test was used to compare RD rates in predicted highand low-risk groups. Results: In the TCGA and Tothill datasets, 491 and 189 patients, respectively, met the criteria for inclusion. As expected, survival was significantly better for patients with R0 resection compared to those with any RD. We identified 47 probesets, representing 38 different genes, significant in both datasets at 10% FDR. These included probesets for FABP4 and ADH1B, which tracked tightly and showed dynamic ranges N16-fold. For these genes, there is a level of expression above which nearly all patients have RD; the distribution of expression values is roughly bimodal. In the validation cohort, using the top quartile of FABP4 polymerase chain reaction values as a prespecified cutoff, we found 30/35 RD cases in the high-expression group and 54/104 in the low-expression group (P= 0.0002). Our predictive method based on FABP4, therefore, correctly identified a cohort of patients with significantly increased rates of RD in an independent test set. Further examination of the ADH1B results showed that predictions using either ADH1B alone or in combination with FABP4 would have produced similar results. Examining the reasons for RD in the high-risk cohort suggests a preponderance of cases with either innumerable sites of disease or unresectable disease involving vital regions. Conclusions: High FABP4 and ADH1B expression are associated with significantly higher risk of RD in HGSOC patients. Patients with high tumoral FABP4 levels may be candidates for neoadjuvant chemotherapy.

Background: CRM1 is a nuclear export protein which increases cell viability through nuclear export signal-dependent protein exclusion of tumor suppressor proteins. Selective Inhibitor of Nuclear Exprot (SINE) is a novel class of drugs that selectively inhibit CRM1. Here, we tested the biological effects of targeting CRM1. Methods and results: To explore the efficacy of SINE CRM1 anatgonists in ovarian cancer, we tested the selective small molecule KPT-185(in vitro) against human ovarian cancer cell lines. KPT-185 reduced cell viability in multiple (A2780, A2780CP20, IGROV-1 and SKOV3) human ovarian cancer cell lines with IC50 levels ranging from 0.2 - 0.75μM. Nuclear localization and significantly increased expression of tumor suppressor proteins (e.g., p53, p21 and Fox03a) was noted in response to KPT-185 treatment. Moreover, synergistic reduction in cell viability was noted with topotecan, cisplatin and liposomal doxorubicin (interaction index was 0.45, 0.78 and 0.59, respectively). Using the A2780 in vivo orthotopic ovarian cancer model, oral administration of KPT-330 (in vivo) monotherapy at 20mg/kg twice a week resulted in 90% tumor reduction (p Conclusion: CRM1 inhibitor is a potent inhibitor of ovarian cancer growth and merits additional development. Citation Format: Takahito M. Miyake, Sunila Pradeep, Behrouz Zand, Heather J. Dalton, Yunfei Wen, Guillermo N. Armaitz Pena, Michael Kauffman, Dilara McCauley, Sharon Shacham, John E. Wiktorowicz, Anil K. Sood. Therapeutic targeting of CRM1 in 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 5541. doi:10.1158/1538-7445.AM2013-5541

Objective: Members of the Eph/Ephrin family are known to play seminal roles in cancer growth and metastasis in many solid tumors; however, a comprehensive analysis has not been carried out. We analyzed the clinical relevance of all Eph/Ephrin family members in endometrial cancer. Methods: Level 3 data from The Cancer Genome Atlas (TCGA) was utilized to assess the clinical significance of all 14 Eph receptors available in the database (EphA1-8, EphA10, EphB1-4, and EphB6), as well as EphrinA1-A5 and EphrinB1-B3. We examined gene methylation status and copy number changes in these tumors. Clinical information extracted included age, tumor histology, tumor grade, stage, estrogen(ER)/progesterone(PR) receptor status, and overall survival. Results: A total of 320 samples were available for analysis. Increased EphA2, EphA4, EphB1, and EphB2 expression was significantly correlated with poorer overall survival (p=0.03, p=0.04, p=0.01, and p=0.008, respectively) and lack of ER and PR expression (r=-0.34, p=2.25e−6; r-0.24, p=0.0007; r=-0.22, p=0.002; r=-0.46, p=2.38e−11, respectively). Compared to endometrioid histology, serous histology was associated with increased expression of EphA2 (p 63 years had greater tumoral expression of EphA4 (p Conclusions: Concordant with emerging roles for the Eph receptor and Eprhins in driving malignant biology, increased EphA2, EphA4, EphB1, and EphB2 expression, as well as EphrinA3 and EphrinA5 expression, is predictive of poor patient outcome and constitutes attractive targets for biological therapies. Citation Format: Heather J. Dalton, Cristina Ivan, Chad V. Pecot, Rajesha Rupamaiole, Behrouz Zand, Justin Bottsford-Miller, Wei Hu, Alpa M. Nick, Robert L. Coleman, Anil K. Sood. An integrated analysis of the eph/ephrin family: implications in endometrial 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 4034. doi:10.1158/1538-7445.AM2013-4034