Early Detection Of Ovarian Cancer Research Articles

Abstract 2302: Methylation profiling of circulating tumor DNA for the detection of ovarian cancer

Abstract Introduction: Ovarian cancer (OC), often diagnosed at an advanced stage, lacks effective screening strategy. It is the most lethal gynecological cancer, with a 5-year survival ranging from 35-50% depending on the subtype. A clear correlation between stage and survival has been demonstrated; therefore, early detection represents the best hope for mortality reduction. This study interrogates the potential of ctDNA methylation status as an early diagnostic marker for OC. Methods: Panel Design: Public data sets such as TCGA and GEO databases (tumor=4,014, normal=352) and in-house generated functional methylome (5.5 million CpG sites, 50X) sequencing data (n=561) were used to profile methylation alterations in 12 common cancer types. The panel covers 225,369 CpG sites, spanning ~3.5Mb of human genome. Deep targeted bisulfite sequencing (ELSA-seq) was performed on plasma samples of 126 Chinese ovarian cancer patients, 59 patients with benign ovarian diseases, including but not limited to endometrial cyst, ovarian cyst and serous cystadenoma and 49 age-matched asymptomatic females to interrogate their methylation statuses with an average sequencing depth of 1,000x. The plasma samples were collected and processed from two centers independently, and randomly assigned into the training (n=123) or validation group (n=113) at 1:1 ratio. Samples that failed quality control specifications were removed from the downstream analysis and the diagnostic information was not disclosed for the validation group until all analyses were completed. Results: First, ovarian tissues samples from 22 OC (FIGO stage II-III) patients, 11 ovarian benign diseases patients and 11 healthy donors were profiled to derive markers that can differentiate the 3 groups. In this study, 45,753 OC-specific differentially methylated CpG sites/loci (DMLs) were selected using modified wald-test with an adjusted p-value <0.05 and fold-change>2, and subsequently grouped into 3,160 differentially methylated blocks (DMBs) based on the co-methylation levels and genomic distances of adjacent CpG sites. Blood samples from 63 patients with OC (4 stage I, 6 stage II and 53 stage III), 33 patients with benign ovarian disease and 27 healthy individuals were obtained to train the diagnostic classification model using a support vector machine (SVM)-based machine learning classifier. Subsequently, iterated 5-fold cross-validation were performed to gain a robust estimation of model performance, achieving a specificity of 91% for benign ovarian diseases and sensitivity of 25%, 50% and 87% for stage I, II and III OC, respectively at a specificity of 93% and area under curve (AUC) of 91.3%. The model was subsequently validated in a hold-out cohort, consisting of 22 healthy individuals, 26 patients with benign ovarian disease and 63 stage I-III OC patients with comparable clinical characteristics as the patients in the training cohort. The model yielded a specificity of 93% for benign disease and a sensitivity of 84% for OC at a specificity of 96% in the validation cohort, suggesting its robustness. Conclusion: Our findings demonstrated the potential of ctDNA methylation profiling, which can effectively distinguish ovarian cancer, benign diseases and healthy, to serve as a diagnostic tool in ovarian cancer early detection. The general concept can be further extended to other types of cancer diagnostics. Citation Format: Ning Li, Yu Zhang, Xin Zhu, Zhenjing Zhang, Jiayue Xu, Bingsi Li, Han Han-Zhang, Fujun Qiu, Shuai Fang, Hao Liu, Zhihong Zhang. Methylation profiling of circulating tumor DNA for the detection of ovarian cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2302.

Association of gBRCA1/2 mutation locations with ovarian cancer risk in Japanese patients from the CHARLOTTE study.

Whether germline (g) breast cancer susceptibility gene (BRCA) mutations are located within or outside the ovarian cancer cluster region (OCCR) (1380‐4062 bp for gBRCA1, and between 3249‐5681 bp and 6645‐7471 bp for gBRCA2) may influence risk variations for ovarian cancers. This ad hoc analysis of the CHARLOTTE epidemiological study in Japan assessed the distribution of gBRCA1/2 mutations in patients with newly diagnosed ovarian cancer, and investigated an association between gBRCA1/2 mutation locations and ovarian cancer risk. Differences in patient background and clinical characteristics in subgroups stratified by gBRCA1/2 mutation locations were also evaluated. We analyzed the data of 93 patients (14.7%) from the CHARLOTTE study who were positive for gBRCA1/2 mutations. After excluding 16 cases with L63X founder mutation, 28 (65.1%) of gBRCA1 mutations were within the OCCR. Of 30 gBRCA2 mutations, 15 (50.0%) were within the OCCR. Of 27 patients (one patient excluded for unknown family history) with gBRCA1 mutations located in the OCCR, 11 (40.7%) had a family history of ovarian cancer; the proportion of patients with a family history of ovarian cancer and gBRCA1 mutations outside the OCCR was lower (13.3%). Sixty percent of patients with gBRCA1 mutations outside the OCCR had a family history of breast cancer; the proportion of patients with a family history of breast cancer and gBRCA1 mutations within the OCCR was relatively lower (33.3%). Understanding the mutation locations may contribute to more accurate risk assessments of susceptible individuals and early detection of ovarian cancer among gBRCA mutation carriers.

Women who develop ovarian cancer show an increase in serum calcium and a decrease in serum albumin. A longitudinal study in the Janus Serum Bank Cohort

BackgroundOvarian cancer is associated with high serum calcium and low serum albumin in clinical and epidemiologic studies. Whether high calcium and low albumin predispose to ovarian cancer or reflect existing cancer is unclear. ObjectiveTest the hypothesis that serum calcium increases and serum albumin decreases in women who develop ovarian cancer. MethodsTwo hundred and four women donated sera to the Janus Serum Bank in Norway pre- and post-diagnosis of ovarian cancer, donations separated by approximately 14 years. We measured calcium and albumin in these sera and calculated the albumin-corrected calcium. Sera were adjusted for patient age and storage time. ResultsPost-diagnosis, mean age- and storage-adjusted calcium increased, from 2.53 to 2.68 mmol/L (p < .001). Mean age- and storage-adjusted, albumin-corrected calcium increased from 2.3 to 2.7 mmol/L (p < .001). Conversely, mean age- and storage-adjusted albumin decreased, from a mean of 51.3 to 40.9 g/L (p < .001). Significant changes were observed in women with early stage and metastatic cancer. ConclusionsThese data support the hypothesis that calcium and albumin are serum biomarkers of extant ovarian cancer. Longitudinal changes in calcium and albumin may be useful in ovarian cancer early detection.

MicroRNA-193b regulates human ovarian cancer cell growth via targeting STMN1.

Ovarian cancer is the eighth most common malignancy among women worldwide. Ovarian cancer exhibits no obvious symptoms in the early stage of tumorigenesis and currently, no effective methods for the early detection and treatment of ovarian cancer have been established. Therefore, the identification of novel targets is critical to the early diagnosis and clinical treatment of ovarian cancer. microRNAs (miRs) are small non-coding RNAs, which serve an important biological role in a number of physiological processes and in oncogenesis. Previous studies have reported that miRNA-193b is dysregulated in a variety of types of human cancer. However, the roles of miRNA-193b in human ovarian cancer has not been determined. The present study investigated the roles of miRNA-193b in human ovarian cancer cells. Reverse transcription-quantitative PCR results indicated that the expression of miRNA-193b in ovarian cancer cells was significantly down-regulated compared with non-malignant cells. Cell counting kit-8 results indicated that the up-regulation of miRNA-193b inhibited ovarian cancer cell proliferation and induced ovarian cancer cell apoptosis. The present study also indicated that stathmin 1 (STMN1) was a direct target of miRNA-193b, and the up-regulation of miRNA-193b significantly decreased the expression of STMN1 in ovarian cancer cells. In conclusion, the results demonstrated that miRNA-193b serves as a tumor suppressor in human ovarian cancer by inhibiting cell proliferation and inducing cell apoptosis. Therefore, the assessment of miRNA-193b may provide insight into a novel diagnostic biomarker and potential therapeutic target for patients with ovarian cancer.

Cu isotope ratios are meaningful in ovarian cancer diagnosis

BackgroundOvarian cancer diagnosis is currently based on imaging and circulating CA-125 concentrations with well-known limits to sensitivity and specificity. New biomarkers are required to complement CA-125 testing to increase effectiveness. Increases in sensitivity of isotopic separation via multi collector inductively coupled plasma-mass spectrometry have recently allowed highly accurate measurement of copper (Cu) isotopic variations. Studies in breast cancer patients have revealed changes of serum copper isotopic composition demonstrating the potential for development as a cancer biomarker. Evaluating 65Cu/63Cu ratios (δ65Cu) in serum samples from cancer patients has revealed a strong correlation with cancer development. In this study blood samples from forty-four ovarian cancer patients, and 13 ovarian biopsies were investigated. ResultsHere we demonstrate that changes in Cu isotopes also occurs in ovarian cancer patients. Copper composition determined by multiple collector inductively coupled plasma mass spectrometry revealed that the copper isotopic ratio δ65Cu in the plasma of 44 ovarian cancer patient cohort was significantly lower than in a group of 48 healthy donors, and indicated that serum was enriched for 63Cu. Further analysis revealed that the isotopic composition of tumour biopsies was enriched for 65Cu compared with adjacent healthy ovarian tissues. ConclusionsWe propose that these changes are due to increase lactate and Cu transporter activities in the tumour. These observations demonstrate that, combined with existing strategies, δ65Cu could be developed for use in ovarian cancer early detection.

Abstract IA10: Traditional and modern approaches for prevention and early detection

Abstract Conventionally, serum CA-125 measurements and ultrasonography have been used to try to detect ovarian carcinoma at early stage. These methods have not been proven to result in early-stage detection and seem to convey little survival benefit. It is now widely accepted that such methods are not justified for widespread use in cancer screening, and the US Preventative Services Task Force recommends against their use. CA-125 and human epididymis protein 4 (HE4) are involved in ovarian tumorigenesis, overexpressed by malignant epithelial cells, and found in patient serum. Increased levels of these proteins as well as others in patient ascites, and uterine fluid, as compared to serum, suggest that their detection may be enhanced by measurements taken from within the uterine cavity or other locations more proximal to the ovaries and fallopian tube where tumors most commonly originate. The knowledge that most high-grade serous carcinomas originate from the distal fallopian tube creates an opportunity for new screening and prevention approaches that were previously impractical when the site of origin was thought to be the ovarian surface epithelium. The fallopian tube can be accessed via a transvaginal/transuterine route, and the fallopian tubes are not necessary for normal biologic function once childbearing is complete. Based on the hypothesis that serous tubal intraepithelial carcinoma (STIC) is the precursor of most high-grade serous carcinomas (HGSCs), innovative studies aimed at providing a realistic clinical method for reducing the burden of ovarian cancer are under way. In this presentation we will discuss the latest knowledge regarding approaches for prevention and early detection of ovarian cancer. We will discuss the role of precursor lesions in the development and metastatic spread of invasive ovarian cancers. We will review ongoing and planned clinical trials of surgical prevention and discuss missed opportunities for chemoprevention. We will also discuss more innovative approaches to early detection through minimally invasive access to the distal fallopian tube and nanotechnology for highly sensitive protein detection. The approaches and others hold great promise to transform the paradigm of early ovarian cancer detection from one of traditional serum-based biomarkers to one that incorporates the current concepts of ovarian cancer origins, proximal measurements of tissue-based biomarkers, and novel circulating biomarkers. Citation Format: Douglas A. Levine. Traditional and modern approaches for prevention and early detection [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research; 2019 Sep 13-16, 2019; Atlanta, GA. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(13_Suppl):Abstract nr IA10.

Developing an Epigenetic Biomarker for Early Detection of Ovarian Cancer

Objectives: Given the difficulty in early detection of ovarian tumors, and the negative impact that late detection has on patient survival rates, there is a vital need for early detection strategies. There is growing interest in immune cells and their surveillance and regulation of cancer growth. The objective of this study was to compare the gene expression and epigenetic patterns in white blood cells isolated from patients with ovarian cancer to age-matched women with pelvic masses without ovarian cancer.

Abstract A13: DNA methylation of SOX1 and HOXA9 as a biomarker for early detection of ovarian cancer in cell-free DNA

Abstract Background: Accumulated evidence revealed that aberrant CpG island hypermethylation plays significant roles in carcinogenesis that can serve as a promising target for molecular detection in body fluids. Ovarian cancer remains the most lethal gynecologic malignancy and is characterized by few early symptoms, late stage presentation, and resulting poor survival, which thereby renders challenge to its prompt diagnosis. Despite a myriad of attempts at early diagnosis of ovarian cancer, this clinical aim still remains a major challenge. To date, no single biomarker is able to accurately detect early OC in either tissue or body fluid. Aberrant circulating DNA methylation patterns provide highly specific cancer signals. Methods: To develop a DNA methylation-based screening assay for early diagnosis of ovarian cancer, we quantitatively assessed the promoter methylation of DAPK1, HOXA9, RASSF1A, and SOX1 genes in 105 ovarian cancer and 30 non-neoplastic ovarian specimens by means of a high-throughput quantitative, real-time PCR-based technique (MethyLight) and clonal bisulfite sequencing. The best-performing gene cassette was further evaluated for their methylation status in cell-free DNA from serum of matched 35 ovarian cancer patients and 20 normal controls. Area under the ROC (Receiver-operator characteristic) curve was used to evaluate the discriminatory performance of both individual and combined gene panel. Results: RASSF1A, HOXA9, DAPK1, and SOX1 displayed a methylation frequency of 72.72, 82.67, 68.0, and 80.0%, respectively, in tissue samples. We identified DNA methylation of HOXA9 and SOX1 as the best discriminator between cancer and non-neoplastic tissue. In the multiplex assay, the combined panel of HOXA9 and SOX1 achieved a sensitivity of 88.0% with a specificity of 86.7%, when either or both of the gene promoters showed methylation. These genes appear to have great potential to be evaluated for their methylation level in cell-free DNA as a noninvasive diagnostic marker for early diagnosis of ovarian cancer. In cell-free DNA, this novel panel achieved a sensitivity of 62.86.0% and a specificity of 95% with an area under the ROC curve of 0.81 for discriminating OC samples from normal control. The results of MethyLight were concordant with those of clonal bisulfite sequencing. Conclusion: Our findings revealed the potential of epigenetic biomarker for early detection of OC in cell-free DNA as a minimally invasive tool. Here, we have established that promoter hypermethylation can be detected in serum cell-free DNA from patients with early as well as advanced-stage ovarian cancer. Based on the multiplex MethyLight assay, study will focus on analyzing the clinical utility of HOXA9 and SOX1 as methylation biomarkers. These findings further underline the potential of multiplexing individual markers into a panel that helps to achieve higher sensitivity and specificity than individual marker and provides robustness to a noninvasive early screening test. Citation Format: Alka Singh, Jaydeep Aravindbhai Badarukhiya, Sameer Gupta, Manisha Sachan. DNA methylation of SOX1 and HOXA9 as a biomarker for early detection of ovarian cancer in cell-free DNA [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr A13.

Abstract A060: [Advocate Abstract] Pathways to ovarian cancer

Abstract Ovarian cancer has common symptoms but how easily are they ignored in the African American community? How is the diagnosis shared amongst family members, particularly amongst females in the immediate family? Is there a cultural stigma that prohibits African American women from being specific about the type of cancer they've been diagnosed with? Can answers to these questions result in earlier detection for the next generation of ovarian cancer survivors? I will explore how and why symptoms can be ignored by age groups from 30 to 65 and the role the gynecologist can play in earlier detection of ovarian cancer. I will lead a discussion on the stigma of cancer in the black community and why African American women tend not to disclose the specificity of their cancer, nor encourage female family members to test for BRCA1. Note: This abstract was not presented at the conference. Citation Format: Kimberly Richardson. [Advocate Abstract] Pathways to ovarian cancer [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr A060.

Abstract D087: Racial/ethnic differences in ovarian cancer incidence and mortality among adult women in the United States, 2001-2016

Abstract Background: Ovarian cancer is the fifth most common cause of cancer death among US women. Assessing racial/ethnic differences in ovarian cancer incidence and mortality will provide important information regarding improving efforts of prevention, early detection, and treatment in population groups that need to be targeted. Methods: We included adult women diagnosed with ovarian cancer ≥ ages 20 from the National Program for Cancer Registries and Surveillance, Epidemiology, and End Results Incidence Incidence–-U.S. Cancer Statistics 2001–2016 database. Incidence of ovarian cancer was age- adjusted to the U.S. standard population. Joinpoint analyses were used to assess incidence trend and calculate annual percentage change (APC). Hazard ratios (HRs) and 95% CIs were estimated from the Cox proportional hazard models for comparing differences in 5-year survival probability across races/ethnicities, after controlling for age at diagnosis and county of residence. Results: Among adult women ≥ 20 years old, ovarian cancer incidence decreased among Hispanics, non-Hispanic Whites, and non-Hispanic Blacks, from 9.4 per 100,000 in 2001 to 6.9 per 100,000 in 2016 (APC -1.8, 95% confidence interval (CI) -2.2 - -1.5) in Hispanics, from 11.0 per 100,000 in 2001 to 7.6 per 100,000 in 2016 (APC -2.1, 95% CI -2.3 - -1.9) in non-Hispanic Whites, and from 8.0 per 100,000 in 2001 to 6.6 per 100,000 in 2016 (APC -1.3, 95% CI -1.6 - -1.0) in non-Hispanic Blacks. No joinpoints were found among those racial/ethnic groups. Among ovarian cancer patients, observed 5-year ovarian-cancer-specific survival was 51.6% (95% CI 50.4%-52.8%) in Hispanics, 44.7% (95% CI 44.2%-45.1%) in non-Hispanic Whites, and 37.4% (95% CI 36.1%-38.8%) in non-Hispanic Blacks (HR for dying from ovarian cancer, Whites vs. Blacks 0.68, 95% CI 0.66-0.71; Hispanics vs. Blacks 0.73, 95% CI 0.69-0.76). Similar patterns were also observed for 5-year overall survival and 5-year relative survival among adult ovarian patients. Conclusions: Ovarian cancer incidence decreased across all three racial/ethnic groups from 2001 to 2016. Although Non-Hispanic Blacks had the lowest ovarian cancer incidence, they had the lowest survival rate. More efforts are needed to focus on this underserved population to target early detection and treatment of ovarian cancer. Citation Format: Fangjian Guo, Abbey B. Berenson, Yong-Fang Kuo. Racial/ethnic differences in ovarian cancer incidence and mortality among adult women in the United States, 2001-2016 [abstract]. In: Proceedings of the Twelfth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2019 Sep 20-23; San Francisco, CA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl_2):Abstract nr D087.

A Systematic Review on Cost-effectiveness Studies Evaluating Ovarian Cancer Early Detection and Prevention Strategies.

Ovarian cancer imposes a substantial health and economic burden. We systematically reviewed current health-economic evidence for ovarian cancer early detection or prevention strategies. Accordingly, we searched relevant databases for cost-effectiveness studies evaluating ovarian cancer early detection or prevention strategies. Study characteristics and results including quality-adjusted life years (QALY), and incremental cost-effectiveness ratios (ICER) were summarized in standardized evidence tables. Economic results were transformed into 2017 Euros. The included studies (N = 33) evaluated ovarian cancer screening, risk-reducing interventions in women with heterogeneous cancer risks and genetic testing followed by risk-reducing interventions for mutation carriers. Multimodal screening with a risk-adjusted algorithm in postmenopausal women achieved ICERs of 9,800-81,400 Euros/QALY, depending on assumptions on mortality data extrapolation, costs, test performance, and screening frequency. Cost-effectiveness of risk-reducing surgery in mutation carriers ranged from cost-saving to 59,000 Euros/QALY. Genetic testing plus risk-reducing interventions for mutation carriers ranged from cost-saving to 54,000 Euros/QALY in women at increased mutation risk. Our findings suggest that preventive surgery and genetic testing plus preventive surgery in women at high risk for ovarian cancer can be considered effective and cost-effective. In postmenopausal women from the general population, multimodal screening using a risk-adjusted algorithm may be cost-effective.

MiR-200b/200a/429 Cluster Stimulates Ovarian Cancer Development by Targeting ING5.

Ovarian cancer is the second most common gynaecological malignancy, and microRNAs (miRNAs) play important role in the cancer development. Here, we found that the level of miR-200b/200a/429 was significantly increased in serum and tumor tissues of patients with stage-I ovarian cancer. Consistent with these results, we detected increased expression levels of miR-200b/200a/429 in ovarian cancer cell lines compared with the human nontumorigenic ovarian epithelial cell line T80. The overexpression of miR-200b/200a/429 in T80 cells stimulated proliferation and caused their growth in soft agar and tumor formation in nude mice. Furthermore, we determined that miR-200b/200a/429 targets inhibitor of growth family 5 (ING5) and that the overexpression of ING5 can block miR-200b/200a/429-induced T80 cell transformation and tumorigenesis. Our findings suggest that miR-200b/200a/429 may be a useful biomarker for the early detection of ovarian cancer and that miR-200b/200a/429 significantly contributes to ovarian cancer development through ING5.

Development of a Targeted Ultrasound Microbubble for Early Detection of Ovarian Cancer

Background and ObjectiveOvarian cancer is the deadliest gynecologic malignancy. Although curable in up to 90% of patients diagnosed during early‐stage disease, most patients are not diagnosed until late‐stage, at which point, the overall 5‐year survival rate is less than 25%. This disparity clearly shows that early detection is a key factor for improving patient survival. However, there is no accurate screening tool available, and currently utilized diagnostic modalities lack specificity for early disease. Transvaginal ultrasound, the imaging gold standard for routine examination of ovarian masses, has many advantages that make it an ideal diagnostic tool, such as its low cost, safety, and sensitivity. Despite its advantages, ultrasound is lacking in specificity, as it is limited in its ability to distinguish between benign and malignant ovarian masses. This study seeks to address this deficiency via a novel technique known as targeted ultrasound. Targeted ultrasound involves the use of micron‐sized (~1–4μm) gas‐filled bubbles stabilized by an outer shell to which targeting molecules, such as antibodies, are conjugated. Microbubbles are injected intravenously, where they travel systemically to bind to their target. Upon confrontation with ultrasound waves, they create a unique ultrasound backscatter, indicating the presence of the target. Due to their micron‐scale size, microbubbles are too large to exit the vascular space and enter surrounding tissues; therefore, targets must be expressed on endothelial cells lining the luminal surface of blood vessels. Previous studies have shown tissue factor (TF) to be a unique, tumor‐specific marker expressed in a number of different tumor types, including ovarian cancer. Although not expressed on normal, healthy vascular endothelial cells (VECs), TF expression has been shown on tumor‐associated VECs. Additionally, TF is associated with tumor angiogenesis ‐ an early event in tumorigenesis. This unique expression profile makes TF an attractive target for early cancer detection via targeted ultrasound. The objective of this study was to develop a TF‐targeted ultrasound microbubble for early detection of ovarian cancer.MethodsTF expression on murine and human ovarian cancer tissues was examined using western blotting and immunohistochemistry. Targeted microbubbles were made by conjugating an anti‐TF antibody to the outer shell of microbubbles via streptavidin‐biotin chemistry. Their binding efficacy and specificity was validated using in vitro cell binding assays.ResultsTF expression was shown on ovarian tumor‐associated VECs in patient and murine tissues. TF‐targeted microbubbles, with the ability to detect and bind to TF‐expressing cells in vitro, were developed.ConclusionsBased upon these findings, we anticipate that in vivo translation of TF‐targeted microbubbles will show effective detection of tumor VECs in a murine model of ovarian cancer. Development of a targeted microbubble against a unique tumor marker, such as TF, would have significant implications towards increasing the number of early‐stage ovarian cancer diagnoses and ultimately, help save the lives of more women affected by this disease.

Detection of aberrant methylation of HOXA9 and HIC1 through multiplex MethyLight assay in serum DNA for the early detection of epithelial ovarian cancer.

Accumulated evidence revealed that aberrant CpG island hypermethylation plays an important role in carcinogenesis which can serve as a promising target for molecular detection in body fluids. Despite a myriad of attempts to diagnose ovarian cancer (OC) at an early stage, this clinical aim remains a major challenge. To date, no single biomarker is able to accurately detect early OC in either tissue or body fluid. Aberrant DNA methylation patterns in circulating DNA provide highly specific cancer signals. In our study, we establish a novel panel of methylation-specific genes for the development of a TaqMan based qPCR assay to quantify methylation levels. We analyzed promoter methylation of homeobox A9 (HOXA9) and hypermethylated in cancer 1 (HIC1) quantitatively in 120 tissue samples and in 70 matched serum cell-free DNA (CFDNA) of cancerous and noncancerous samples by MethyLight assay. HOXA9 and HIC1 methylation occurred in 82.3 and 80.0% of OC tissue samples in singleplex assay, thereby confirming that methylation was highly cancer-specific. When either or both gene promoter showed methylation, the sensitivity was 88.2% with a specificity of 88.6% in tissue samples. The combined sensitivity for this novel marker panel in serum CFDNA was 88.9% (area under the curve [AUC] = 0.95). In contrast, no hypermethylation was observed in serum from matched cancer-free control women. Our results confirm the elevated performance of novel epigenetic marker panel (HOXA9 and HIC1) when analyzed in tissue and matched serum samples. Our findings reveal the potential of this biomarker panel as a suitable diagnostic serum biomarker for early screening of OC.

Ovarian cancer cells with CD133+ phenotype is more resistant against Ngai Bun Boesenbergia pandurata extract than original ovarian cancer cells

Introduction: Ovarian cancer is one of the most common cancers in women. Due to the difficulty in early detection and treatment of ovarian cancer, many research studies and clinical trials have been developed to discover more efficient therapies. Besides Western medicine, traditional medicine has gained increased interest as a research field with potential to lead to the production of marketable therapeutic products. With the diversity of tropical plants in Asia, traditional medicine has been very popular and has served as a traditional therapy for generations. The Ngai bun (Boesenbergia pandurata) root is used not only as a food spice but also in ethnomedicine. This study aimed to compare the anti-tumor activity of Boesenbergia pandurata root extract against ovarian cancer cells and CD133+ ovarian cancer cells that were enriched from the original ovarian cancer cells.&#x0D; Methods: Crude extract of Boesenbergia pandurata roots were prepared in two kinds of solvents (methanol and chloroform). The ovarian cancer cells OVP-10 were used in this study. The population of CD133+ ovarian cancer cells (CD133+OVP-10) were sorted from the OVP-10 cancer cells. Both OVP-10 cells and CD133+OVP-10 cells were treated with these crude extracts. Adiposederived stem cells (ADSCs) were used as control normal cells for all assays. The anti-tumor activity of extracts were evaluated based on the IC50 values.&#x0D; Results: Based on the IC50 index, the chloroform extract had an anti-tumor activity higher than that of methanol extract, on both OVP-10 and CD133+OPV-10 cells (IC50 of methanol and chloroform extracts were 330.1 +/- 16.9 ug/mL and 246.5 +/- 21.2 ug/mL, respectively, for OVP-10 cells; IC50 of methanol and chloroform extracts were 411.8 +/- 83.7 ug/mL and 307 +/- 9.2 ug/mL respectively, for CD133+OVP-10 cells). The results also showed that CD133+OVP-10 cells were more resistant to chloroform extract than were OVP-10 cells (307 +/- 9.2 mg/mL vs. 246.5 +/- 21.2 mg/mL, respectively, for CD133+OVP-10 vs. OVP-10 cells, p &lt; 0.05).&#x0D; Conclusion: The chloroform extract of Boesenbergia pandurata roots displayed strong antitumor activity against ovarian cancer cells OVP-10 and CD133+OVP-10; the latter cells were found to be more resistant than the original ovarian cancer cells.&#x0D;

Identification of a four-gene methylation biomarker panel in high-grade serous ovarian carcinoma.

Background The lack of effective biomarkers for the screening and early detection of ovarian cancer (OC) is one of the most pressing problems in oncogynecology. Because epigenetic alterations occur early in the cancer development, they provide great potential to serve as such biomarkers. In our study, we investigated a potential of a four-gene methylation panel (including CDH13, HNF1B, PCDH17 and GATA4 genes) for the early detection of high-grade serous ovarian carcinoma (HGSOC). Methods For methylation detection we used methylation sensitive high-resolution melting analysis and real-time methylation specific analysis. We also investigated the relation between gene hypermethylation and gene relative expression using the 2-ΔΔCt method. Results The sensitivity of the examined panel reached 88.5%. We were able to detect methylation in 85.7% (12/14) of early stage tumors and in 89.4% (42/47) of late stage tumors. The total efficiency of the panel was 94.4% and negative predictive value reached 90.0%. The specificity and positive predictive value achieved 100% rates. Our results showed lower gene expression in the tumor samples in comparison to control samples. The more pronounced downregulation was measured in the group of samples with detected methylation. Conclusions In our study we designed the four-gene panel for HGSOC detection in ovarian tissue with 100% specificity and sensitivity of 88.5%. The next challenge is translation of the findings to the less invasive source for biomarker examination, such as plasma. Our results indicate that combination of examined genes deserve consideration for further testing in clinical molecular diagnosis of HGSOC.

Feasibility, patient compliance and acceptability of ovarian cancer surveillance using two serum biomarkers and Risk of Ovarian Cancer Algorithm compared to standard ultrasound and CA 125 among women with BRCA mutations

ObjectiveWe assessed the feasibility, patient acceptability of and compliance of a new surveillance strategy for ovarian cancer surveillance in women with BRCA mutations, based on assessments of serum CA125 and HE4 every 4 months (Risk of Ovarian Cancer Algorithm (ROCA) arm), compared to Standard of Care (SOC) surveillance with CA125 blood tests and pelvic ultrasounds every 6 months. MethodsWomen were recruited 6/13/16–9/11/17 from an integrated health care system in California for this non-randomized prospective cohort study. Women were invited to participate in a novel serum biomarker surveillance strategy using ROCA or they could opt to be in the standard of care control arm with ultrasound and CA 125 every 6 months. Outcomes assessed included compliance, self-reported distress using the Impact of Event Scale (IES) and cancer anxiety using the Cancer Worry Scale. ResultsThere were 159 women in the ROCA arm and 43 in the SOC arm. Overall, compliance was higher in the ROCA arm (83.2%) than in SOC (51.9%), p < 0.0001. Based on the IES, ROCA arm women reported less feelings about intrusion and avoidance at 12 months compared to baseline; the difference approached significance for intrusion (7.6% vs 4.1% severe, p = 0.057) and was statistically significant for avoidance (20.8% vs 9.9% severe, p = 0.034). ConclusionsThis pilot demonstrated that compliance was high with blood tests performed every four months for ovarian cancer surveillance. Moreover, ROCA women had lower stress scores over time than SOC women. Given the lack of clinical utility and poor compliance shown with traditional ultrasound and CA125 tests, further investigation is warranted of longitudinal biomarker surveillance for early detection of ovarian cancer.

Evaluation of paraneoplastic antigens reveals TRIM21 autoantibodies as biomarker for early detection of ovarian cancer in combination with autoantibodies to NY-ESO-1 and TP53.

The majority of ovarian cancer cases are diagnosed at an advanced stage with poor prognosis. This study evaluates autoantibodies against tumor antigens to identify candidate biomarkers for early detection of ovarian cancer in women at increased risk. To assess the immunoreactivity of paraneoplastic antigens and tumor associated antigens with high-grade serous ovarian cancer (HGSOC) samples. Five paraneoplastic antigens along with three tumor-associated antigens were evaluated with HGSOC patient serum samples. Validation screening was performed with n= 164 serum samples consisting of: 50 late stage HGSOC, 14 early stage HGSOC, 50 benign ovarian cyst, and 50 healthy control samples on ELISA and western blot. The four markers TRIM21, NY-ESO-1, TP53, and PAX8 were evaluated on a second validation serum set, n= 150. TRIM21 achieved the highest sensitivity in the first validation screening of 33% with 100% specificity. Combining TRIM21 with NY-ESO-1, TP53, and PAX8 provided 67% sensitivity with 94% specificity, and 56% sensitivity at 98% specificity. These four markers resulted in 46% sensitivity with 98% specificity in the second validation cohort; TRIM21 achieved the highest individual sensitivity of 36%. Autoantibodies to TRIM21, NY-ESO-1, and TP53 may complement CA125 in screening of women at genetic risk for ovarian cancer.

Reducing Ovarian Cancer Mortality Through Early Detection: Approaches Using Circulating Biomarkers.

More than two-thirds of all women diagnosed with epithelial ovarian cancer (EOC) will die from the disease (>14,000 deaths annually), a fact that has not changed considerably in the last three decades. Although the 5-year survival rates for most other solid tumors have improved steadily, ovarian cancer remains an exception, making it the deadliest of all gynecologic cancers and five times deadlier than breast cancer. When diagnosed early, treatment is more effective, with a 5-year survival rate of up to 90%. Unfortunately, most cases are not detected until after the cancer has spread, resulting in a dismal 5-year survival rate of less than 30%. Current screening methods for ovarian cancer typically use a combination of a pelvic examination, transvaginal ultrasonography, and serum cancer antigen 125 (CA125), but these have made minimal impact on improving mortality. Thus, there is a compelling unmet need to develop new molecular tools that can be used to diagnose early-stage EOC and/or assist in the clinical management of the disease after a diagnosis, given that more than 220,000 women are living with ovarian cancer in the United States and are at risk of recurrence. Here, we discuss the state of advancing liquid-based approaches for improving the early detection of ovarian cancer.See all articles in this Special Collection Honoring Paul F. Engstrom, MD, Champion of Cancer Prevention.

Evaluation of miR-22 and miR-21 as diagnostic biomarkers in patients with epithelial ovarian cancer.

Among the 11 most common cancers, ovarian cancer is the fifth leading cause of death after lung, breast, colorectal and pancreatic cancer. Although diagnosis of ovarian cancer in early stages is followed by various successful treatments, no accurate and reliable method is available at present. Currently microRNAs are being explored as signature biomarkers for early detection of various types of cancer. However little information is available on expression and correlation of microRNA in ovarian cancer. In this study, we have chosen two microRNA on the basis of their altered frequency in epithelial ovarian cancer cases. The main objective of this study is to evaluate the expression of microRNA-22 and microRNA-21 along with various clinicopathological parameters. Expression level of microRNA-22 and microRNA-21 in different stages and subtypes of epithelial ovarian carcinoma has been analyzed to find out its role as a potential diagnostic biomarker. Present study has been conducted in the serum of 80 epithelial ovarian cancer patients and 80 age matched healthy women. Quantitative real time PCR was used to compare the expression of miR-22 and miR-21 between the cases and control groups. Statistical analysis showed 7.85-fold increase in miR-21 expression and 2.1-fold reduction in miR-22 expression of ovarian cancer patients. Increased serum level of miR-21 in ovarian cancer patients and decreased level of miR-22 has been correlated with advanced international federation of gynecology and obstetrics (FIGO) stage and histological sub types of epithelial ovarian cancer. Serous ovarian carcinoma was the most common cancer in the present study. Calculated fold change value for miR-21 was 3.98 and - 2.86 for miR-22 in serous ovarian cancer. Fold change value in the miR-21 expression in advanced stage was 6.29 and 4.25 in early stage. Whereas lower calculated fold change was observed for miR-22 in advanced stage than in early stage (- 2.16). Present study revealed up-regulated expression of miR-21 and down regulated expression of miR-22 in the serum of epithelial ovarian cancer patients. Both of these could be validated as good diagnostic biomarkers for early detection of ovarian cancer.