Late-stage Epithelial Ovarian Cancer Research Articles

Diagnosing and staging epithelial ovarian cancer by serum glycoproteomic profiling.

There is a need for diagnostic tests for screening, triaging and staging of epithelial ovarian cancer (EOC). Glycoproteomics of blood samples has shown promise for biomarker discovery. We applied glycoproteomics to serum of people with EOC or benign pelvic masses and healthy controls. A total of 653 analytes were quantified and assessed in multivariable models, which were tested in an independent cohort. Additionally, we analyzed glycosylation patterns in serum markers and in tissues. We identified a biomarker panel that distinguished benign lesions from EOC with sensitivity and specificity of 83.5% and 90.1% in the training set, and of 86.7 and 86.7% in the test set, respectively. ROC analysis demonstrated strong performance across a range of cutoffs. Fucosylated multi-antennary glycopeptide markers were higher in late-stage than in early-stage EOC. A comparable pattern was found in late-stage EOC tissues. Blood glycopeptide biomarkers have the potential to distinguish benign from malignant pelvic masses, and early- from late-stage EOC. Glycosylation of circulating and tumor tissue proteins may be related. This study supports the hypothesis that blood glycoproteomic profiling can be used for EOC diagnosis and staging and it warrants further clinical evaluation.

H3K27me3 in Diffuse Midline Glioma and Epithelial Ovarian Cancer: Opposing Epigenetic Changes Leading to the Same Poor Outcomes.

Histone post-translational modifications modulate gene expression through epigenetic gene regulation. The core histone H3 family members, H3.1, H3.2, and H3.3, play a central role in epigenetics. H3 histones can acquire many post-translational modifications, including the trimethylation of H3K27 (H3K27me3), which represses transcription. Triple methylation of H3K27 is performed by the histone methyltransferase Enhancer of Zeste Homologue 2 (EZH2), a component of the Polycomb Repressive Complex 2. Both global increases and decreases in H3K27me3 have been implicated in a wide range of cancer types. Here, we explore how opposing changes in H3K27me3 contribute to cancer by highlighting its role in two vastly different cancer types; (1) a form of glioma known as diffuse midline glioma H3K27-altered and (2) epithelial ovarian cancer. These two cancers vary widely in the age of onset, sex, associated mutations, and cell and organ type. However, both diffuse midline glioma and ovarian cancer have dysregulation of H3K27 methylation, triggering changes to the cancer cell transcriptome. In diffuse midline glioma, the loss of H3K27 methylation is a primary driving factor in tumorigenesis that promotes glial cell stemness and silences tumor suppressor genes. Conversely, hypermethylation of H3K27 occurs in late-stage epithelial ovarian cancer, which promotes tumor vascularization and tumor cell migration. By using each cancer type as a case study, this review emphasizes the importance of H3K27me3 in cancer while demonstrating that the mechanisms of histone H3 modification and subsequent gene expression changes are not a one-size-fits-all across cancer types.

Hyperthermic intraperitoneal chemotherapy for treating ovarian cancer.

Hyperthermic intraperitoneal chemotherapy (HIPEC) has been used for decades as a treatment for disseminated abdominal malignancies, including pseudomyxoma peritonei and peritoneal mesothelioma. HIPEC had been used to treat recurrent ovarian cancer only when curative options were lacking, but new data indicate that it may be suitable as a primary treatment for patients with late-stage epithelial ovarian cancer, increasing the recurrence-free time interval and improving quality of life. However, treatment can be challenging because of patient trauma from surgery, combined with the toxicity and high temperature of the chemotherapeutic agents. This article reviews HIPEC and its targeted intraoperative and postoperative management.

MR imaging of epithelial ovarian cancer: a combined model to predict histologic subtypes.

To compare the performance of clinical features, conventional MR image features, ADC value, T2WI, DWI, DCE-MRI radiomics, and a combined multiple features model in predicting the type of epithelial ovarian cancer (EOC). In this retrospective analysis, 61 EOC patients were confirmed by histology. Significant features (p < 0.05) by multivariate logistic regression were retained to establish a clinical model, conventional MRI morphological model, ADC model, and traditional model. The radiomics model included FS-T2WI, DWI, and DCE-MRI, and also, a multisequence model was established. A total of 1070 radiomics features of each sequence were extracted; then, univariate analysis and LASSO were used to select important features. Traditional models were combined with a combined radiomics model to establish a mixed model. The predictive performance was validated by receiver operating characteristic curve (ROC) analysis, calibration curve, and decision curve analysis (DCA). A stratified analysis was conducted to compare the differences between the combined radiomics model and the traditional model in identifying early- and late-stage EOC. Traditional models showed the highest performance (AUC = 0.96). The performance of the mixed model (AUC = 0.97) was not significantly different from that of the traditional model. The calibration curve showed that the traditional model had the highest reliability. Stratified analysis showed the potential of the combined radiomics model in the early distinction of the two tumor types. The traditional model is an effective tool to distinguish EOC type I/II. Combined radiomics models have the potential to better distinguish EOC types in early FIGO stage disease. • The combined radiomics model resulted in a better predictive model than that from a single sequence model. • The traditional model showed higher classification accuracy than the combined radiomics model. • Combined radiomics models have the potential to better distinguish EOC types in early FIGO stage disease.

Clinicopathologic factors associated with short-term survival in advanced epithelial ovarian cancer

Background: Epithelial ovarian cancer (EOC) is the most lethal gynaecological malignancy, and there is a deficiency of information in the literature on the early recognition of short-term survivor (STS). This study aimed to identify the clinicopathological factors associated with STS in late-stage EOC and to establish a predictive model to identify STS. Methods: Selected patients with International Federation of Gynecology and Obstetrics (FIGO) stage III or IV EOC were included in the study, and a retrospective analysis was performed. The characteristics of the patients who survived not more than 2 years (STS) were compared to those who survived at least 2 years (defined as long-term survivors, LTS). Binary logistic regression and receiver operating characteristic curve (ROC) were used to identify the independent prognostic factors associated with EOC and assess the predictive accuracy for STS. Results: We identified 254 patients with advanced EOC including 57 STS and 197 LTS. A univariate analysis revealed that STS had a tendency to have omental metastasis and larger tumor size, to be platinum resistant, to have non-serous histology, to undergo suboptimal cytoreduction, to have comorbidity, and to undergo primary chemotherapy less than 6 courses. Binary regression analysis revealed that tumor size (P=0.033), platinum resistance (P Conclusions: Tumor size, omental metastasis, platinum resistance, non-serous histology, and number of primary chemotherapy are predictors associated with STS when controlling other confounding factors. Tumor size and omental metastasis may be considered novel, important prognostic factors for advanced EOC patients. Platinum resistance was the most important prognosticator for STS; hence, more work is needed for the early identification and treatment of these EOC patients.

Clinicopathologic factors associated with short-term survival in advanced epithelial ovarian cancer

BackgroundEpithelial ovarian cancer (EOC) is the most lethal gynaecological malignancy, and there is a deficiency of information in the literature on the early recognition of short-term survivor (STS). This study aimed to identify the clinicopathological factors associated with STS in late-stage EOC and to establish a predictive model to identify STS.MethodsSelected patients with International Federation of Gynecology and Obstetrics (FIGO) stage III or IV EOC were included in the study, and a retrospective analysis was performed. The characteristics of the patients who survived not more than 2 years (STS) were compared to those who survived at least 2 years (defined as long-term survivors, LTS). Binary logistic regression and receiver operating characteristic curve (ROC) were used to identify the independent prognostic factors associated with EOC and assess the predictive accuracy for STS.ResultsWe identified 254 patients with advanced EOC including 57 STS and 197 LTS. A univariate analysis revealed that STS had a tendency to have omental metastasis and larger tumor size, to be platinum resistant, to have non-serous histology, to undergo suboptimal cytoreduction, to have comorbidity, and to undergo primary chemotherapy less than 6 courses. Binary regression analysis revealed that tumor size (P=0.033), platinum resistance (P<0.001), non-serous histology (P=0.048) and number of primary chemotherapy (P=0.028) were significant independent predictors of STS. A developed predictive model using these predictors had an AUC =0.831; platinum resistance alone had an AUC =0.732.ConclusionsTumor size, omental metastasis, platinum resistance, non-serous histology, and number of primary chemotherapy are predictors associated with STS when controlling other confounding factors. Tumor size and omental metastasis may be considered novel, important prognostic factors for advanced EOC patients. Platinum resistance was the most important prognosticator for STS; hence, more work is needed for the early identification and treatment of these EOC patients.

Changes in mRNA/protein expression and signaling pathways in in vivo passaged mouse ovarian cancer cells.

The cure rate for late stage epithelial ovarian cancer (EOC) has not significantly improved over several decades. New and more effective targets and treatment modalities are urgently needed. RNA-seq analyses of a syngeneic EOC cell pair, representing more and less aggressive tumor cells in vivo were conducted. Bioinformatics analyses of the RNA-seq data and biological signaling and function studies have identified new targets, such as ZIP4 in EOC. Many up-regulated tumor promoting signaling pathways have been identified which are mainly grouped into three cellular activities: 1) cell proliferation and apoptosis resistance; 2) cell skeleton and adhesion changes; and 3) carbohydrate metabolic reprograming. Unexpectedly, lipid metabolism has been the major down-regulated signaling pathway in the more aggressive EOC cells. In addition, we found that hypoxic responsive genes were at the center stage of regulation and detected functional changes were related to cancer stem cell-like activities. Moreover, our genetic, cellular, biochemical, and lipidomic analyses indicated that cells grown in 2D vs. 3D, or attached vs. suspended had dramatic changes. The important clinical implications of peritoneal cavity floating tumor cells are supported by the data proved in this work. Overall, the RNA-seq data provide a landscape of gene expression alterations during tumor progression.

The impact of DNA damage response gene polymorphisms on therapeutic outcomes in late stage ovarian cancer.

Late stage epithelial ovarian cancer has a dismal prognosis. Identification of pharmacogenomic markers (i.e. polymorphisms) to stratify patients to optimize individual therapy is of paramount importance. We here report the retrospective analysis of polymorphisms in 5 genes (ATM, ATR, Chk1, Chk2 and CDK12) involved in the cellular response to platinum in a cohort of 240 cancer patients with late stage ovarian cancer. The aim of the present study was to evaluate associations between the above mentioned SNPs and patients’ clinical outcomes: overall survival (OS) and progression free survival (PFS). None of the ATM, ATR, Chk1 and Chk2 polymorphisms was found to significantly affect OS nor PFS in this cohort of patients. Genotype G/G of CDK12 polymorphism (rs1054488) predicted worse OS and PFS than the genotype A/A-A/G in univariate analysis. The predictive value was lost in the multivariate analysis. The positive correlation observed between this polymorphism and age, grade and residual tumor may explain why the CDK12 variant was not confirmed as an independent prognostic factor in multivariate analysis.The importance of CDK12 polymorphism as possible prognostic biomarker need to be confirmed in larger ovarian cancer cohorts, and possibly in other cancer population responsive to platinum agents.

Abstract 5054: Role and signaling of desmosomal DSP and PKP1 in ovarian cancer

Abstract Identification of driver genes/proteins in EOC tumorigenic processes is critical to increase the cure rate for late stage epithelial ovarian cancer (EOC), which has not been in improved over several decades. We have developed a highly aggressive EOC cell line (ID8-P1 or P1) from ID8-P0 (without in vivo passage, or P0) through in vivo passage in C57BL6 mice. Two genes encode important desmosome components, desmoplakin (Dsp) and plakophilin 1 (Pkp1) were found to be significantly over-expressed in P1 vs. P0 cells, as well as human EOC cells. DSP is an obligate component of functional desmosomes that anchors intermediate filaments to desmosomal plaques. Pkp1 encodes a member of the arm-repeat (armadillo) and plakophilin (PKP) gene families. Our understanding of the functions of desmosomes in cancer is just emerging and the role of desmosomes in EOC is totally unknown. Our data showed the novel functions of DSP in anoikis-resistance and SRC activation using shRNA-mediated DSP-knocked down (KD) stable cell lines. In addition, we found that ID8-P1 cells formed spheroids and P0 cells tended to be single cells when they were cultured in suspension. KD-Dsp dissociated spheroids, suggesting that desmosomes play an important role in spheroid formation, which may be directly related to anoikis-, drug-resistance, and/or the cancer stem cell (CSC) concepts. Moreover, we found that enhanced glycolysis was involved in the increased anoikis-resistance of ID8-P1 cells and an inhibitor of glycolysis, shikonin (SHK) was a potent anoikis-resistance inhibitor and P1 cells gained more drug-resistance, when compared to P0 cells, with the IC50s were 1.7 and 7.4 μM for P0 and P1 cells, respectively. Similarly, the IC50s were 2.4 and 8.1 μM for human EOC PE01 and PE04 cells, respectively. PE01 and PE04 cell lines are identified to represent high-grade serous ovarian cancer (HGSC), which were derived from the same patient before (PE01) and after (PE04) the onset of drug resistance to cis-platinum (CDDP), chlorambucil, and 5-fluorouracil. PP2 (a SRC nhibitor) and SHK were able to dissociate the cell-cells aggregates/spheroids in EOC cells. The previously unknown role of DSP and/or PKP1 in EOC may be related to their signaling activities outside desmosomes. Cells staining of PKP1 protein showed its cytosolic and perinuclear location, providing supporting and proof-of-concept data. Together, our data have suggested the novel links among DSP, PKP1, anoikis-resistance, SRC activation, cell-cell-aggregation, and drug-resistance. These data provide a basis for developing new modalities for EOC treatment. Citation Format: Yan Xu, Qingchun Cai. Role and signaling of desmosomal DSP and PKP1 in ovarian cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 5054.

Abstract POSTER-CTRL-1211: Haptoglobin identified within ovarian cyst fluid as an accurate intraoperative diagnostic biomarker for epithelial ovarian cancer

Abstract Introduction: Epithelial ovarian cancer (EOC) is the most lethal gynaecological malignancy, with a poor prognosis and low survival rate; most cases are diagnosed at a late stage due to the fact that symptoms at early stages are usually non-specific in nature. There is currently no screening method proven to be effective in improving the outcome of EOC patients; existing biomarkers for EOC, such as CA-125, generally suffer from a lack of specificity in early stage disease, which is the ideal time for therapeutic intervention. Aim: We hypothesized that a single protein biomarker within the ovarian cyst fluid (OCF) could be identified, isolated, characterized and validated for application in a point-of-care device (POC), which could subsequently be used in operating theaters for triage for frozen section (FS). Methods: We screened the OCF proteome by mass-spectrometry (MALTI-TOF/MS), confirmed the identity of the protein by western blot and SELDI immunocapture analysis. Next, demonstrated using tissue microarray (TMA) that cellular expression of haptoglobin varied in normal, benign and malignant ovarian tissue. We developed a simple ELISA and a rapid colorimetric assay that allowed semi-quantification of OCF haptoglobin intraoperatively. Finally, we validated a point-of-care test kit to accurately identify EOC using OCF with a higher predictive value than can be achieved using RMIs, and an equivalent accuracy to intraoperative FS. Results: The OCF haptoglobin concentration in benign tumors was 0.70±0.09 mg/ml compared to 6.22±0.53 mg/ml and 6.57±0.65 mg/ml in early- and late-stage EOCs, respectively (P&amp;lt;0.0001). We sourced a rapid colorimetric assay and determined the accuracy of this assay on archived OCF samples (n=124). The accuracy of this rapid colorimetric assay was similar to that of our in-house sandwich ELISA with sensitivity of 97.3% (95% CI: 84.2-99.9%), specificity of 92.0% (95% CI: 83.6-96.4%), PPV of 83.7% (95% CI: 68.7-92.7%) and NPV of 98.8% (95% CI: 92.4-99.9%). In an early phase clinical trial of the POC device, we prospectively tested 50 OCF samples real-time in the operation theater away from the surgeon’s view and the POC device tester blinded to the type of surgery performed and diagnosis. Visual calls from POC device were recorded and compared to histology. The POC device had an accuracy of 82% with sensitivity of 70% (95% CI: 35.1-91.9%), specificity of 85% (95% CI: 69.5-93.8%), PPV of 53.8% (95% CI: 26.1-79.6%), and NPV of 91.9% (95% CI: 77-97.9%). Conclusion: This is the first study whereby an intraoperative tumor marker has been utilised in the differentiation between benign and malignant ovarian lesions. Its accuracy suggests that it could be utilised as a replacement or an adjunct to frozen section, particularly in situations where histopathological expertise is scarce. Citation Format: Mahesh Choolani, Loganath Annamalai, Lin Liu, Khalil Razvi, Changqing Zhao, Gregory Rice, Aniza P Mahyuddin, Arijit Biswas, Jerry Chan, Narasimhan Kothandaraman. Haptoglobin identified within ovarian cyst fluid as an accurate intraoperative diagnostic biomarker for epithelial ovarian cancer [abstract]. In: Proceedings of the 10th Biennial Ovarian Cancer Research Symposium; Sep 8-9, 2014; Seattle, WA. Philadelphia (PA): AACR; Clin Cancer Res 2015;21(16 Suppl):Abstract nr POSTER-CTRL-1211.

Abstract A48: M2 macrophage inhibition reverses vascular leaks that cause malignant ascites in late-stage epithelial ovarian cancer

Abstract Introduction: Malignant ascites is a common symptom of epithelial ovarian cancer (EOC) that greatly reduces the quality of life for patients and has no good treatment options that target the source of the problem. Our study aims to find the cause of ascites in order for us to better know how to safely reverse this pathology. Experimental Procedures: Murine ID8 EOC tumor cells were implanted intraperitoneally or orthotopically into female C57BL/6 mice, and bioluminescent imaging was used to track the progress of the cancer. GW2580, a very selective colony stimulating factor 1 receptor (CSF1R) inhibitor, was given to mice for two weeks once ascites appeared. Flow cytometry, histology, and endothelial cell permeability assays were used to analyze the effects of blocking macrophages in the model. Flow cytometry and endothelial cell permeability assays were also used to analyze and characterize EOC patient ascites samples. Results: CSF1R inhibition blocked specifically M2 macrophages from the ascites and dramatically reduced ascites volume, leaving mice less anemic. When M2 macrophages numbers were reduced in the ascites, CD8 T cell numbers increased, causing the ascites microenvironment to be less immunosuppressive. Peritoneal vasculature that had grown disorganized and leaky became normalized with treatment. Ascites sera from GW2580-treated mice protected against endothelium permeability, while control mouse sera caused the endothelium to become leaky. This data agrees with our finding that patient ascites sera cause more endothelium permeability if more macrophages were present in the ascites. We also saw macrophage and myeloid cell depletion in the tumor and systemically with GW2580 treatment, along with a consistent reduction in tumor burden. Conclusions: M2 macrophages are key players that cause ascites through deregulating peritoneal vasculature. The leaky vessels are normalized once these macrophages are physically and functionally blocked from the ascites microenvironment. This finding suggests that macrophage inhibition could be a powerful tool for ascites management. VEGF inhibitors are currently in clinical trials for malignant ascites treatment with mixed, sometimes deadly, results. Targeting macrophages, the source of many vessel-deregulating proteins, might make it more difficult for resistance to build up than when just one protein, such as VEGF, is blocked, and CSF1R inhibitors have been shown to be safe and well-tolerated in the clinic. This novel approach toward ascites and EOC management should be further studied. Future work will focus on finding the specific factors that caused the vessel deregulation and normalization, looking at functional differences between macrophage from control and GW2580-treated ascites, seeing if endothelial cell-macrophage direct interaction may have any effects on the vasculature, and performing this study in genetic and human models of EOC. Citation Format: Diana Moughon, Huanhuan He, Shiruyeh Schokrpur, Ziyue Jiang, Madeeha Yaqoob, John David, Luisa Iruela-Arispe, Oliver Dorigo, Lily Wu. M2 macrophage inhibition reverses vascular leaks that cause malignant ascites in late-stage epithelial ovarian cancer. [abstract]. In: Abstracts: AACR Special Conference on Cellular Heterogeneity in the Tumor Microenvironment; 2014 Feb 26-Mar 1; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2015;75(1 Suppl):Abstract nr A48. doi:10.1158/1538-7445.CHTME14-A48

Large-scale profiling of metabolic dysregulation in ovarian cancer.

Ovarian cancer is the leading cause of death in gynecologic malignancies. Profiling of endogenous metabolites has potential to identify changes caused by cancer and provide inspiring insights into cancer metabolism. To systematically investigate ovarian cancer metabolism, we performed metabolic profiling of 448 plasma samples related to epithelial ovarian cancer (EOC) based on ultra-performance liquid chromatography mass spectrometry in both positive and negative modes. These unbiased metabolomic profiles could well distinguish EOC from benign ovarian tumor (BOT) and uterine fibroid (UF). Fifty-three metabolites were identified as specific biomarkers for EOC, and this is the first report of piperine, 3-indolepropionic acid, 5-hydroxyindoleacetaldehyde and hydroxyphenyllactate as metabolic biomarkers of EOC. The AUC values of these metabolites for discriminating EOC from BOT/UF and early-stage EOC from BOT/UF were 0.9100/0.9428 and 0.8385/0.8624, respectively. Meanwhile, our metabolites were able to distinguish early-stage EOC from late-stage EOC with an AUC of 0.8801. Importantly, analysis of dysregulated metabolic pathways extends our current understanding of EOC metabolism. Metabolic pathways in EOC patients are mainly characterized by abnormal phospholipid metabolism, altered l-tryptophan catabolism, aggressive fatty acid β-oxidation and aberrant metabolism of piperidine derivatives. Together, these metabolic pathways provide a foundation to support cancer development and progression. In conclusion, our large-scale plasma metabolomics study yielded fundamental insights into dysregulated metabolism in ovarian cancer, which could facilitate clinical diagnosis, therapy, prognosis and shed new lights on ovarian cancer pathogenesis.

The serum glycome to discriminate between early-stage epithelial ovarian cancer and benign ovarian diseases.

Epithelial ovarian cancer (EOC) is the sixth most common cause of cancer deaths in women because the diagnosis occurs mostly when the disease is in its late-stage. Current diagnostic methods of EOC show only a moderate sensitivity, especially at an early-stage of the disease; hence, novel biomarkers are needed to improve the diagnosis. We recently reported that serum glycome modifications observed in late-stage EOC patients by MALDI-TOF-MS could be combined as a glycan score named GLYCOV that was calculated from the relative areas of the 11 N-glycan structures that were significantly modulated. Here, we evaluated the ability of GLYCOV to recognize early-stage EOC in a cohort of 73 individuals comprised of 20 early-stage primary serous EOC, 20 benign ovarian diseases (BOD), and 33 age-matched healthy controls. GLYCOV was able to recognize stage I EOC whereas CA125 values were statistically significant only for stage II EOC patients. In addition, GLYCOV was more sensitive and specific compared to CA125 in distinguishing early-stage EOC from BOD patients, which is of high relevance to clinicians as it is difficult for them to diagnose malignancy prior to operation.

Utility of Progranulin and Serum Leukocyte Protease Inhibitor as Diagnostic and Prognostic Biomarkers in Ovarian Cancer

Epithelial ovarian cancer (EOC) is the fifth leading cause of cancer-related death in females and leading gynecologic cause of cancer-related death. Despite the identification of a number of serum biomarkers, methods to identify early-stage disease and predict prognosis remain scarce. We have evaluated two biologically connected serum biomarkers, serum leukocyte protease inhibitor (SLPI) and progranulin (PGRN). Two-hundred frozen plasma samples were acquired from the Mayo Clinic Biospecimen Repository for Ovarian Cancer Research. Samples were obtained from 50 patients with benign conditions, 50 with American Joint Committee on Cancer (AJCC) stage I and II EOC, and 100 with AJCC stage III and IV EOC. Samples were obtained before surgical resection of a mass and were analyzed for absolute levels of SLPI and PGRN using ELISA assays. Receiver-operator characteristic curves were generated for SLPI and PGRN. Median follow-up was 48 months. Absolute levels of SLPI were significantly elevated in patients with EOC compared with benign disease and predicted the presence of EOC (AUC of 0.812; P = 0.04); SLPI remained elevated in the subset of patients with normal CA-125. PGRN levels were not significantly increased in patients with early-stage or late-stage EOC as a whole, but an increase in PGRN levels was associated with decreased overall survival in advanced EOC. SLPI levels are elevated in EOC, and SLPI shows promise as a diagnostic biomarker for patients with both elevated and normal CA-125 levels. An increase in PGRN is associated with decreased overall survival. SLPI is elevated in EOC and warrants investigation in a screening study in women at risk for EOC.

Comparative effectiveness of treatments for recurrent ovarian cancer.

e16551 Background: Traditionally, recurrent ovarian cancer has been treated with chemotherapy. However, increasingly, secondary cytoreductive surgery (SCS) is being performed. We undertook this study to compare survival between SCS vs chemotherapy vs both or neither to treat recurrent ovarian cancer. Methods: Using SEER-Medicare 1997-2007, we identified women age 66+ years with primary late stage epithelial ovarian cancer with fee-for-service Medicare treated with surgery and chemotherapy for their primary cancer who experienced a recurrence. We defined recurrent cancer using claims for chemotherapy, hospice or SCS following a 6 month treatment free window after primary treatment. Survival time is calculated from time of first recurrence. We conducted parametric and semi-parametric multivariate survival analyses to assess comparative treatment survival rates and factors affecting survival. We use propensity score weighting methods to obtain robust estimates of comparative effectiveness. To address the risk that a fraction of patients classified by our algorithms as non-recurrent are, indeed, recurrent, we use finite mixture models. Results: Of the 1,623 women with recurrent cancer, 72% received secondary chemotherapy while 15% receive both chemotherapy and surgery. &lt;2% receive secondary surgery and 12% received hospice with no active treatment. Univariate analyses show that the mean survival time after first recurrence of ovarian cancer is 1,357 days. Multivariate survival models controlling for patient demographics, cancer type and comorbidities show that, compared to patients who received both surgery and chemotherapy, those who received only SCS (-569 days) as well as those who received only chemotherapy (-381 days) had significantly lower survival times. Survival time is negatively associated with black race and age at recurrence and positively associated with date of reoccurrence. Patients with greater comorbidities have significantly shorter survival times. Conclusions: Patients with recurrent ovarian cancer treated with both secondary surgery and chemotherapy survive longer than patients treated with either chemotherapy or surgery. Women who are Black, or older at time of recurrence have worse survival.

Abstract PL05-04: Epigenetic marks as biomarkers and their mechanisms of maintenance as targets in ovarian cancer.

Abstract PL05-04: Epigenetic marks as biomarkers and their mechanisms of maintenance as targets in ovarian cancer.

Measuring the intra-individual variability of the plasma proteome in the chicken model of spontaneous ovarian adenocarcinoma

The domestic chicken (Gallus domesticus) has emerged as a powerful experimental model for studying the onset and progression of spontaneous epithelial ovarian cancer (EOC) with a disease prevalence that can exceed 35% between 2 and 7 years of age. An experimental strategy for biomarker discovery is reported herein that combines the chicken model of EOC, longitudinal plasma sample collection with matched tissues, advanced mass spectrometry-based proteomics, and concepts derived from the index of individuality (Harris, Clin Chem 20: 1535-1542, 1974). Blood was drawn from 148 age-matched chickens starting at 2.5 years of age every 3 months for 1 year. At the conclusion of the 1 year sample collection period, the 73 birds that remained alive were euthanized, necropsied, and tissues were collected. Pathological assessment of resected tissues from these 73 birds confirmed that five birds (6.8%) developed EOC. A proteomics workflow including in-gel digestion, nanoLC coupled to high-performance mass spectrometry, and label-free (spectral counting) quantification was used to measure the biological intra-individual variability (CV(W)) of the chicken plasma proteome. Longitudinal plasma sample sets from two birds within the 73-bird biorepository were selected for this study; one bird was considered "healthy" and the second bird developed late-stage EOC. A total of 116 proteins from un-depleted plasma were identified with 80 proteins shared among all sample sets. Analytical variability (CV(A)) of the label-free proteomics workflow was measured using a single plasma sample analyzed five times and was found to be ≥CV(W) in both birds for 16 proteins (20%) and in either bird for 25 proteins (31%). Ovomacroglobulin (ovostatin) was found to increase (p < 0.001) over a 6 month period in the late-stage EOC bird providing an initial candidate protein for further investigation.

Abstract B5: Haptoglobin proved to be a novel biomarker for intraoperative triage of epithelial ovarian cancer at early stage

Abstract Introduction: Epithelial ovarian cancer (EOC) is the leading cause of gynecological cancer-related deaths worldwide, and the prognosis can be greatly improved if cancer is detected at early stage. Intra-operative suspicion of malignancy for women going for ovarian cystectomy is critical for triage of patients for the most suitable surgical procedures. In this study, we aimed to evaluate the diagnostic accuracy of haptoglobin level in ovarian cyst fluid as a potential biomarker for intra-operative triage of EOC in both local (Singapore) and regional (Indonesia and Vietnam) centers and to explore its possible clinical applications. Methods: We measured haptoglobin concentration in ovarian cyst fluid samples, that were carefully collected during surgery without spillage, from 113 benign tumors, 24 early stage and 31 late stage cancers in Department of Obstetrics and Gynaecology, National University Hospital in Singapore and 6 regional centers in Southeast Asia from 2004–2009, using an in-house sandwich enzyme-linked immunosorbent assay (ELISA). We also tested the feasibility of using a rapid colorimetric assay (Phase™ range haptoglobin assay, Tridelta Development Limited, Wicklow, Ireland) which measures haptoglobin in 5 minutes and could be potentially used in the operation theater for intra-operative cancer triage and compared it with ELISA and frozen section results. Results: Our data indicated that cyst fluid haptoglobin level was significantly elevated in both early and late stage EOCs (6.51±2.2 and 6.00±1.9 mg/ml, respectively) as compared to benign tumors (0.83±0.9 mg/ml, p &amp;lt;0.001). It was also proven to be an independent marker from the gold standard serum CA125 as there was absence of linear correlation between the two proteins in early stage (r=-0.109, p=0.735), late stage EOCs (r=0.216, p=0.299) and EOCs of all stages (r=-0.039, p=0.796). It showed significantly superior overall diagnostic accuracy [area under the curve (AUC) 0.998, 95% CI 0.995–1.002] over serum CA125 (AUC 0.925, 95% CI 0.874–0.977) and risk of malignancy index (RMI) (AUC 0.914, 95% CI 0.859–0.969) in discriminating between benign and malignant tumors by the receiver operating characteristic (ROC) curve analysis (p&amp;lt;0.05). Using 2.85 mg/ml as the cut-off level, cyst fluid haptoglobin achieved sensitivity of 98.2% (95% CI 89.0–99.9%), specificity of 97.3% (95% CI 91.9–99.3%), positive predictive value (PPV) of 94.7% (95% CI 84.5–98.6%) and negative predictive value (NPV) of 99.1% (95% CI 94.4–100%). A rapid way of measuring haptoglobin level was needed in order to be used for intra-operative diagnosis. Thus a rapid colorimetric assay was employed and the result was almost as good as ELISA. Haptoglobin level measured by the rapid assay was able to separate malignant from benign tumors with sensitivity of 98.1% (95% CI 88.6–99.9%), specificity of 92.1% (95% CI 84.5–96.3%), positive predictive value (PPV) of 86.7% (95% CI 74.9–93.7%) and negative predictive value (NPV) of 93.4% (95% CI 93.4–99.9%). Hence haptoglobin measured by both ELISA and rapid colorimetric assay can used as an alternative to frozen section diagnosis (especially in developing countries where frozen section is not available) or as low-cost triage methods. A point-of-care diagnostic device that employs the principle of the colorimetric assay has been developed and is currently being tested for rapid intra-operative cancer triage. Conclusion: Cyst fluid haptoglobin is a reliable maker for intra-operative diagnosis of EOC at early stage not only in local but also in regional sample sets, with high sensitivity for malignant tumors and a low false-positive rate allowing for optimal surgical procedure to be carried out. Citation Information: Clin Cancer Res 2010;16(7 Suppl):B5

Predictive Value of Aurora-A/STK15 Expression for Late Stage Epithelial Ovarian Cancer Patients Treated by Adjuvant Chemotherapy

To investigate the expression and regulation of the centrosomal kinase Aurora-A/STK15 (AURKA) in epithelial ovarian cancers and to determine the prognostic and predictive value of this marker for patients with late stage epithelial ovarian cancer treated by distinct adjuvant chemotherapies. Archival resection specimens of epithelial ovarian cancers (n=115) and nonneoplastic ovaries (n=28) were analyzed for AURKA mRNA and protein expression by microdissection and quantitative reverse transcriptase-PCR and immunohistochemistry. AURKA DNA copy numbers were measured by fluorescence in situ hybridization in 37 cases. Statistical evaluation was done with respect to clinicopathologic variables, disease-free survival, and overall survival. AURKA mRNA expression was significantly elevated in cancers (P<0.001) and correlated with AURKA protein expression (P=0.0134). Overexpression of AURKA protein was detected in 68 of 107 (63.5%) cases and was linked with increased AURKA DNA copy numbers (P=0.0141) and centromere 20 aneusomy (P=0.0137). Moreover, AURKA overexpression was associated with improved overall survival in optimal debulked patients receiving taxol/carboplatin therapy (n=43, P=0.018). Finally, in an exploratory approach, patients receiving non-taxane-based therapy, AURKA overexpression was predictive for worse overall survival (n=30, P=0.049). AURKA overexpression is seen in the majority of late stage epithelial ovarian cancers, most likely due to increased AURKA DNA copy numbers and/or chromosome 20 aneusomy. Importantly, AURKA overexpression may differentially affect taxane and non-taxane-based adjuvant therapy responses. The study sheds new light on AURKA expression and regulation in epithelial cancers in vivo and specifically shows its value as a clinically relevant marker and as a potential therapeutic target per se.

Oregovomab: profile report

The lifetime risk of ovarian cancer has been estimated to be 1.4% (1 in 70 women), with a 5-year survival rate for all stages of 52% compared with 95% if diagnosed and treated while the disease is still localised.[4] Owing to the poorly defined symptom profile of ovarian cancer, approximately 70% of women are diagnosed with ovarian cancer at stages III and IV with an associated 5-year survival rate of 15–20% and 70% of patients relapse within 6–24 months of second-line treatment,[6] suggesting there is a need for further treatment options. Oregovomab (OvaRex®1) is an immunotherapeutic monoclonal antibody (MAb) with high affinity for the ovarian tumour-associated antigen, cancer 125 (CA125).[1] CA125 is shed from ovarian cancer cells and is elevated in 50% and 80% of patients with stage I and late-stage epithelial ovarian cancer, respectively.[4] Oregovomab induces broad humoral and cellular responses against CA125,[1] and is currently being evaluated as a treatment for patients with late-stage ovarian cancer after completion of first-line therapy. In patients in remission with biochemical relapse, the median time to relapse was greater in oregovomab recipients with an immune response than in nonresponders (35.3 vs 7.5 weeks; p = 0.0011) in a double-blind, placebo-controlled study.[7] A similar study in patients without biochemical relapse also reported that median time to relapse was greater in oregovomab recipients with an immune response than in those who did not respond (13.4 vs 4.7 months; p < 0.0001).[8] In a preliminary integrated analysis of two noncomparative trials in patients in relapse, the median time to disease progression was greater in oregovomab-treated patients with an immune response than in nonresponders (60.0 vs 10.7 weeks; p = 0.0026).[9] The tolerability profile of oregovomab is similar to that of placebo[8] with no treatment-related toxicity reported.[10]