Biomarker For Epithelial Ovarian Cancer Research Articles

Recent Advances in Surface Plasmon Resonance (SPR) Technology for Detecting Ovarian Cancer Biomarkers

Epithelial Ovarian Cancer (EOC) is a leading cause of cancer-related deaths among women, mainly due to a lack of early detection and screening methods. Advanced immunoassay techniques, such as Luminex and proximity extension assay (PEA) technology, show promise in improving EOC detection by utilizing highly sensitive and specific multiplex panels to detect multiple combinations of biomarkers. However, these advanced immunoassay techniques have certain limitations, especially in validating the performance characteristics such as specificity, sensitivity, limit of detection (LOD), and dynamic range for each EOC biomarker within the panel. Implementing multiplexing in point-of-care (POC) biosensors can enhance EOC biomarker detection, with Surface Plasmon Resonance (SPR) being a versatile option among optical biosensors. There is no study on multiplex SPR biosensors specifically tailored for diagnosing EOC. Recent studies have shown promising results in the single detection of EOC biomarkers using SPR, with LOD for cancer antigen 125 (CA125) at 0.01 U/mL−1 and human epididymis protein 4 (HE4) at 1pM. This study proposes a potential roadmap for scientists and engineers in academia and industry to develop a cost effective yet highly efficient SPR biosensor platform for detecting EOC.

Genome-wide expression profiling reveals novel biomarkers in epithelial ovarian cancer

Epithelial ovarian cancer (EOC) is the most aggressive and frequent malignancy detected among women worldwide. The pathophysiology of OC should, therefore be better understood to identify diagnostic, prognostic, and predictive novel biomarkers necessary for early detection, management, and prognostication. In this study, we aimed to investigate transcriptomic landscape and biomarker through RNA-seq data analysis. Further analysis by Protein Protein network identified top 10 Differentially Expressed Genes (DEGs). KEGG pathway enrichment analysis revealed the significant enrichment of DEGs in basal cell carcinoma, cell cycle and FoxO signalling pathway. The RNA-seq results of 10 DEGs were validated by QRT-PCR and TCGA database. Correlation studies were also performed between gene expression and clinical characteristics followed by survival analysis. Finally, 8 DEGs (CDKN1A, BCL6, CDC45, WNT2, TLR5, AQP5) including two novel DEGs (CSN1S1 and NKILA) were identified showing significant correlations with EOC characteristics. These may serve as interesting biomarkers and novel treatment targets and warrant further investigation into the functional outcome of EOC.

Immune checkpoints as potential theragnostic biomarkers for epithelial ovarian cancer.

Epithelial ovarian cancer (EOC) is the leading cause of death associated with gynecologic tumors. EOC is asymptomatic in early stages, so most patients are not diagnosed until late stages, highlighting the need to develop new diagnostic biomarkers. Mediators of the tumoral microenvironment may influence EOC progression and resistance to treatment. To analyze immune checkpoints to evaluate them as theranostic biomarkers for EOC. Serum levels of 16 immune checkpoints were determined in EOC patients and healthy controls using the MILLIPLEX MAP® Human Immuno-Oncology Checkpoint Protein Magnetic Bead Panel. Seven receptors: BTLA, CD40, CD80/B7-1, GITRL, LAG-3, TIM-3, TLR-2 are differentially expressed between EOC and healthy controls. Serum levels of immune checkpoints in EOC patients are positively significantly correlated with levels of their ligands, with a higher significant correlation between CD80 and CTLA4 than between CD28 and CD80. Four receptors, CD40, HVEM, PD-1, and PD-L1, are positively associated with the development of resistance to Taxol-platinum-based chemotherapy. All of them have an acceptable area under the curve (>0.7). This study has yielded a first panel of seven immune checkpoints (BTLA, CD40, CD80/B7-1, GITRL, LAG-3, TIM-3, TLR-2) associated with a higher risk of EOC and a second panel of four immune checkpoints (CD40, HVEM, PD-1, PD-L1) that may help physicians to identify EOC patients who are at high risk of developing resistance to EOC chemotherapy.

The performance of Carbohydrate Antigen 125-Thomsen-nouveau and anti-Müllerian hormone combined with CA125, Human epididymis protein 4 and Risk of Malignancy Algorithm in diagnosis for patients with Epithelial ovarian cancer

ObjectivesWe examined the blood concentrations of Carbohydrate Antigen 125-Thomsen-nouveau (CA125-Tn) and anti-Müllerian hormone (AMH) in epithelial ovarian cancer (EOC) patients to evaluate their potential diagnostic utility together with CA125, human epididymis protein 4 (HE4) and Risk of Malignancy Algorithm (ROMA). Design & methods50 healthy subjects, 45 EOC patients, 22 patients with borderline ovarian tumors (BOT), 21 patients with benign ovarian tumor (BET) and 45 patients with chocolate cyst of ovary (CCO) were studied. Blood levels of CA125, HE4, CA125-Tn and AMH were measured, and the ROMA value was calculated. We compared the differences in the levels of these biomarkers among groups. Additionally, a total of 10 testing strategies were established for comparison to maximize the diagnostic value. ResultsThe levels of CA125, HE4, CA125-Tn and ROMA value were significantly higher in EOC group compared with either the disease control (DC) group (BOT group, BET group and CCO group) or healthy control (HC) group (p < 0.001). In addition, they had better discriminatory performance with an area under the receiver operator characteristic curve (AUC) 0.93; 0.93; 0.93; 0.85, respectively (p < 0.001) compared with the AUC value of AMH 0.67 (p < 0.001). Among all 10 testing strategies, both single-positive of ROMA and double-positive of any 2 markers showed better Youden index (0.82, 0.79, respectively) and kappa value (κ) (0.82, 0.81, respectively). ConclusionsCA125-Tn and AMH can be treated as useful biomarkers of EOC when combined with CA125, HE4 and ROMA, because when any two biomarkers of them are positive, the value of EOC diagnosis is maximized.

Mir-21 and Mir-125b as theranostic biomarkers for epithelial ovarian cancer in Tunisian women.

Ovarian cancer (OC) is the third most common cancer in women and the leading cause of death associated with gynecologic tumors. Because this disease is asymptomatic in the early stages, most patients are not diagnosed until the late stages. This highlights the need for the development of diagnostic biomarkers. MicroRNAs (miRNAs), small non-coding RNAs, are currently being explored as potential biomarkers for the early detection of various malignancies in humans. However, their expression and diagnostic value in OC have not been well studied. the plasma levels of miR-21, miR-200a, miR-200b, miR-200c, miR-205 and miR-125b were determined in epithelial ovarian cancer (EOC) patients and healthy controls by Reverse Transcription Quantitative Realtime Polymerase Chain Reaction (RT-qPCR). The expression levels of the deregulated microRNAs were analysed according to clinical characteristics. It was found that miR-21 and miR-125b were upregulated in EOC compared with healthy controls. Moreover, decreased miR-125b was associated with resistance to platinum-based chemotherapy. Our data suggest that miR-21 and miR-125b in plasma may serve as potential circulating biomarkers for the early detection of EOC. MiR-125b may also be useful for predicting chemosensitivity in EOC patients.

Strategies for data normalization and missing data imputation and consequences for potential diagnostic microRNA biomarkers in epithelial ovarian cancer.

MicroRNAs (miRNAs) are small non-coding RNA molecules regulating gene expression with diagnostic potential in different diseases, including epithelial ovarian carcinomas (EOC). As only a few studies have been published on the identification of stable endogenous miRNA in EOC, there is no consensus which miRNAs should be used aiming standardization. Currently, U6-snRNA is widely adopted as a normalization control in RT-qPCR when investigating miRNAs in EOC; despite its variable expression across cancers being reported. Therefore, our goal was to compare different missing data and normalization approaches to investigate their impact on the choice of stable endogenous controls and subsequent survival analysis while performing expression analysis of miRNAs by RT-qPCR in most frequent subtype of EOC: high-grade serous carcinoma (HGSC). 40 miRNAs were included based on their potential as stable endogenous controls or as biomarkers in EOC. Following RNA extraction from formalin-fixed paraffin embedded tissues from 63 HGSC patients, RT-qPCR was performed with a custom panel covering 40 target miRNAs and 8 controls. The raw data was analyzed by applying various strategies regarding choosing stable endogenous controls (geNorm, BestKeeper, NormFinder, the comparative ΔCt method and RefFinder), missing data (single/multiple imputation), and normalization (endogenous miRNA controls, U6-snRNA or global mean). Based on our study, we propose hsa-miR-23a-3p and hsa-miR-193a-5p, but not U6-snRNA as endogenous controls in HGSC patients. Our findings are validated in two external cohorts retrieved from the NCBI Gene Expression Omnibus database. We present that the outcome of stability analysis depends on the histological composition of the cohort, and it might suggest unique pattern of miRNA stability profiles for each subtype of EOC. Moreover, our data demonstrates the challenge of miRNA data analysis by presenting various outcomes from normalization and missing data imputation strategies on survival analysis.

Serum Exosomes From Epithelial Ovarian Cancer Patients Contain LRP1, Which Promotes the Migration of Epithelial Ovarian Cancer Cell

Ovarian cancer is a gynecological tumor with extremely high mortality and poor prognosis. Exosomes derived from tumor cells contain abundant proteins that may influence tumor metastasis. The purpose of our study was to explore the proteomic profile of serum exosomes from epithelial ovarian cancer (EOC) patients and to find potential diagnostic markers for EOC. We obtained purified exosomes from serum using ultracentrifugation. Migration assay was used to evaluate the effects of exosomes on the migration of EOC cells. Proteomic profile of serum exosomes was analyzed by liquid chromatogram-tandem mass spectrometry. The levels of low-density lipoprotein receptor-related protein 1 (LRP1) in serum and serum exosomes were determined by enzyme-linked immunosorbent assay. Western blot and Immunohistochemistry were used to determine the level of LRP1 in tissues. Moreover, we performed small-interfering RNA-mediated knockdown of LRP1 in EOC cells to obtain SI-LRP1-Exos and SI-NC-Exos. The detailed mechanisms by which exosomal LRP1 affected the migration of EOC cells in vitro and in vivo were also explored. We found that serum exosomes from EOC patients contributed to the migration of EOC cells. The level of serum exosomal LRP1 of EOC patients was significantly upregulated compared with that of healthy volunteers, which was consistent with the result of enzyme-linked immunosorbent assay. We found that exosomal LRP1 regulated the expression of MMP2 and MMP9 through ERK signaling pathway and affected the migration of EOC cells in vitro and in vivo. Therefore, we propose that exosomal LRP1 contributes to the migration of EOC and may act as an important diagnostic and prognostic biomarker of EOC.

Predicting chemoresponsiveness in epithelial ovarian cancer patients using circulating small extracellular vesicle-derived plasma gelsolin

BackgroundResistance to chemotherapy continues to be a challenge when treating epithelial ovarian cancer (EOC), contributing to low patient survival rates. While CA125, the conventional EOC biomarker, has been useful in monitoring patients’ response to therapy, there are no biomarkers used to predict treatment response prior to chemotherapy. Previous work in vitro showed that plasma gelsolin (pGSN) is highly expressed in chemoresistant EOC cell lines, where it is secreted in small extracellular vesicles (sEVs). Whether sEVs from tumour cells are secreted into the circulation of EOC patients and could be used to predict patient chemoresponsiveness is yet to be determined. This study aims to identify if sEV-pGSN in the circulation could be a predictive biomarker for chemoresistance in EOC.MethodsSandwich ELISA was used to measure pGSN concentrations from plasma samples of 96 EOC patients (primarily high grade serous EOC). sEVs were isolated using ExoQuick ULTRA and characterized using western blot, nanoparticle tracking analysis, and electron microscopy after which pGSN was measured from the sEVs. Patients were stratified as platinum sensitive or resistant groups based on first progression free interval (PFI) of 6 or 12 months.ResultsTotal circulating pGSN was significantly decreased and sEV-pGSN increased in patients with a PFI ≤ 12 months (chemoresistant) compared to those with a PFI > 12 months (chemosensitive). The ratio of total pGSN to sEV-pGSN further differentiated these groups and was a strong predictive marker for chemoresistance (sensitivity: 73.91%, specificity: 72.46%). Predetermined CA125 was not different between chemosensitive and chemoresistant groups and was not predictive of chemoresponsiveness prior to treatment. When CA125 was combined with the ratio of total pGSN/sEV-pGSN, it was a significant predictor of chemoresponsiveness, but the test performance was not as robust as the total pGSN/sEV-pGSN alone.ConclusionsTotal pGSN/sEV-pGSN was the best predictor of chemoresponsiveness prior to treatment, outperforming the individual biomarkers (CA125, total pGSN, and sEV-pGSN). This multianalyte predictor of chemoresponsiveness could help to inform physicians’ treatment and follow up plan at the time of EOC diagnosis, thus improving patients’ outcomes.

Analysis of Erythrocyte Sedimentation Rate Order in Epithelial Ovarian Cancer.

Erythrocyte sedimentation rate order (ESRO), platelet-lymphocyte ratio (PLR), red cell distribution width (RDW), and neutrophil-lymphocyte ratio (NLR) are hematological parameters that reflect the presence of biomarkers in epithelial ovarian cancer (EOC). This study evaluated the best haematological parameter in order to distinguish between EOC patients and healthy individuals. There were a total of 33 patients with EOC as subjects treated in Dr. Sardjito hospital, Yogyakarta, Indonesia and 32 volunteer subjects as controls. Curves of receiver operation, area under the curve, specificity, and sensitivity were estimated. To demonstrate EOC's presence, the ESRO was better than the hematological parameters of RDW, PLR, and NLR (AUC = 0.9245, 0.9010, 0.8351, and 0.7457, respectively; sensitivity and specificity = 83.3 and 90.6; 88.9 and 87.5; 100.0 and 68.8; and 94.4 and 53.1, respectively) at the cut-off points 1,125, 44.2, 163.2, and 2,551. Hence, ESRO is a better parameter to indicate the presence of EOC compared to RDW, PLR and NLR.

Identification of two theranostic biomarker panels for epithelial ovarian cancer.

Epithelial Ovarian cancer (EOC) is the leading cause of death associated with gynecologic tumors. Because the disease is asymptomatic in early-stage, the majority of patients are not diagnosed until late stages, highlighting the need for the development of novel diagnostic biomarkers. Mediators of tumoral microenvironment may affect EOC progression and resistance to treatment. Analysis of serum proteins to identify a panel of theranostic biomarkers for EOC. Serum levels of 65 analytes were determined in EOC patients, and healthy controls with the ProcartaPlex Human Immune Monitoring 65-Plex Panel. Twenty-one analytes: 7 cytokines (IFN-γ, IL-12p70, IL-13, IL-18 and TSLP), 7 chemokines (Eotaxin, eotaxin-2, IP-10, BLC, I-TAC, SDF-1α, and fractalkine), 2 growth factors (MMP-1, VEGF-α), and 5 soluble receptors (APRIL, CD40L, TWEAK, CD30 and TNFRII; were significantly differentially expressed between the two groups. ROC curves showed that only seven of them (IL-9, TNF-α, Eotaxin, IP-10, BLC, Fractalkine, and Tweak) had AUC values greater than 0.70 and thus had potential clinical utility. Moreover, five cytokines: IFN-γ, IL-1 β, IL-8, MIP-1β, and TNF-α are positively associated with patients who developed resistance to taxol-platinum-based chemotherapy (CT). This study has revealed a first panel of 7 analytes (IL-9, TNF-α, Eotaxin, IP-10, BLC, Fractalkine and Tweak) that can be used for early detection of EOC and a second panel of five cytokines (IFN-γ, IL-1β, IL-8, MIP-1β, TNF-α) that can help clinicians to identify EOC patients who are at higher risk to develop resistance to CT of EOC.

Identify Biomarkers and Design Effective Multi-Target Drugs in Ovarian Cancer: Hit Network-Target Sets Model Optimizing.

Epithelial ovarian cancer (EOC) is highly aggressive with poor patient outcomes, and a deeper understanding of ovarian cancer tumorigenesis could help guide future treatment development. We proposed an optimized hit network-target sets model to systematically characterize the underlying pathological mechanisms and intra-tumoral heterogeneity in human ovarian cancer. Using TCGA data, we constructed an epithelial ovarian cancer regulatory network in this study. We use three distinct methods to produce different HNSs for identification of the driver genes/nodes, core modules, and core genes/nodes. Following the creation of the optimized HNS (OHNS) by the integration of DN (driver nodes), CM (core module), and CN (core nodes), the effectiveness of various HNSs was assessed based on the significance of the network topology, control potential, and clinical value. Immunohistochemical (IHC), qRT-PCR, and Western blotting were adopted to measure the expression of hub genes and proteins involved in epithelial ovarian cancer (EOC). We discovered that the OHNS has two key advantages: the network's central location and controllability. It also plays a significant role in the illness network due to its wide range of capabilities. The OHNS and clinical samples revealed the endometrial cancer signaling, and the PI3K/AKT, NER, and BMP pathways. MUC16, FOXA1, FBXL2, ARID1A, COX15, COX17, SCO1, SCO2, NDUFA4L2, NDUFA, and PTEN hub genes were predicted and may serve as potential candidates for new treatments and biomarkers for EOC. This research can aid in better capturing the disease progression, the creation of potent multi-target medications, and the direction of the therapeutic community in the optimization of effective treatment regimens by various research objectives in cancer treatment.

Plasma exosomal miR-320d, miR-4479, and miR-6763-5p as diagnostic biomarkers in epithelial ovarian cancer.

Exosomal miRNA had been proved as the promising biomarkers for multiple cancers including epithelial ovarian cancer (EOC). This study aimed to validate the diagnostic accuracy of exosomal miR-320d, miR-4479, and miR-6763-5p for EOC. Exosomes isolated from the plasma by ultracentrifugation were verified using TEM, qNano and western blot. MiRNAs sequencing was used to screen out the differential exosomal miRNAs and miR-320d, miR-4479, and miR-6763-5p were selected as candidates, which were further verified by RT-qPCR in 168 healthy donors and 161 primary EOC patients. Besides, the diagnostic accuracy of these three exosomal miRNAs were evaluated using the receiver operating characteristic curve (ROC). MiRNAs sequencing revealed 95 differential exosomal miRNAs between EOC patients and healthy donors. Subsequently, exosomal miR-320d, miR-4479, and miR-6763-5p were significantly down regulated in EOC patients compared with healthy controls and benign patients. More importantly, these three miRNAs could serve as circulating diagnostics biomarkers for EOC, possessing areas under the curve (AUC) of 0.6549, 0.7781, and 0.6834, respectively. Moreover, these three exosomal miRNAs levels were closely associated with lymph node metastasis, meanwhile exosomal miR-320d and miR-4479 expression was related to tumor stage. Exosomal miR-320d, miR-4479, and miR-6763-5p might serve as potential biomarkers for EOC.

Systematic Pan-Cancer Analysis and Experimental Verification Identify FOXA1 as an Immunological and Prognostic Biomarker in Epithelial Ovarian Cancer.

Epithelial ovarian cancer (EOC) has the lowest survival rate among female reproductive cancers present with symptoms of aggressive malignancies, poor prognosis, drug resistance and postoperative recurrence. The majority of patients with EOC are diagnosed at an advanced stage due to the therapeutic challenges including lack of early diagnosis and effective therapeutic targets for EOC. Pan-cancer analyses were performed to explore the features of forkhead-box (FOX) A1 (FOXA1) using data from TCGA and GTEx databases. R package "clusterprofiler" was used to perform the enrichment analysis of FOXA1 in EOC. Data downloaded from Drug Sensitivity in Cancer (GDSC) database were used to evaluate the association between FOXA1 and antitumor drug sensitivity. In experimental verification, FOXA1 expression was detected using qRT-PCR and western blot assays. Western blot, immunofluorescence staining, and Transwell assays were used to assess the influence of FOXA1 silencing on epithelial-mesenchymal transition (EMT) of EOC cells. We found that FOXA1 was highly expressed in EOC and predicted poorer survival of EOC patients. We observed that FOXA1 expression was positively correlated EMT-related pathways. Through experimental verification, we found the underlying function of FOXA1 to promote EMT in ovarian cancers. The results from western blot, immunofluorescence staining, and Transwell assays showed that FOXA1 silencing impeded the progression of EMT and invasiveness of the cancer cells. Furthermore, CCK-8 and invasion assays suggested that siRNA-FOXA1 attenuated the ability of cancer cells to metastasize and proliferate. Dual-luciferase reporter assays confirmed the binding activity of FOXA1 to the promoter of connective tissue growth factor (CTGF). In addition, we found that FOXA1 was closely correlated immunosuppressive microenvironment of EOC. High FOXA1 expression may contribute to the resistance of many anticancer drugs. Our results predict and validate the function of FOXA1 in promoting EMT and the progression of disease in EOC. Targeting FOXA1 may improve the sensitivity of EOC treatment.

NANOG regulates epithelial-mesenchymal transition via AMPK/mTOR signalling pathway in ovarian cancer SKOV-3 and A2780 cells.

NANOG engages with tumour initiation and metastasis by regulating the epithelial–mesenchymal transition (EMT) in epithelial ovarian cancer (EOC). However, its role in association with pAMPKα, and its clinical significance in EOC have not been elucidated even though AMPK is known to degrade NANOG in various human cancers. Hence, we investigated the role of pAMPKα and its association with NANOG as potential prognostic biomarkers in EOC. Both NANOG and pAMPKα expression were significantly overexpressed in EOCs comparing nonadjacent normal epithelial tissues, benign tissues, and borderline tumours. NANOG overexpression was significantly associated with poor disease‐free survival (DFS) and overall survival (OS), whereas pAMPKα overexpression was associated with good DFS and OS. Importantly, multivariate analysis revealed that the combination of high NANOG and low pAMPKα expression was a poor independent prognostic factor for DFS and was associated with platinum resistance. In ovarian cancer cell lines, siRNA‐mediated NANOG knockdown diminished migration and invasion properties by regulating the EMT process via the AMPK/mTOR signalling pathway. Furthermore, treatment with AMPK activator suppressed expression of stemness factors such as NANOG, Oct4 and Sox2. Collectively, these findings established that the combination of high NANOG and low pAMPKα expression was associated with EOC progression and platinum resistance, suggesting a potential prognostic biomarker for clinical management in EOC patients.

Elevated serum level of human epididymal protein 4 (HE4) predicts poor prognosis in the critically ill with sepsis: A prospective observational cohort study

ObjectiveHuman epididymal protein 4 (HE4) has been widely used as an important clinical tumor biomarker for epithelial ovarian cancer. HE4 has recently been suggested to be an inflammatory biomarker and we hypothesized that the serum HE4 level upon intensive care unit (ICU) admission might predict prognosis in septic patients. We hypothesized that serum HE4 level upon intensive care unit (ICU) admission could predict prognosis in septic patients. MethodsSerum levels of HE4, procalcitonin (PCT), C-reactive protein (CRP), IL-6 and IL-8 were quantified, and sequential organ failure assessment (SOFA) scores were recorded on day one of admission to ICU. The area under the receiver operating characteristic (ROC) curve (AUC) analysis of HE4, IL-6, PCT and SOFA at ICU admission for 28-day mortality was used to evaluate the ability of HE4 in predicting 28-day mortality of sepsis. Multivariate regression analysis was used to identify the independent risk factors for 28-day mortality. ResultsA total of 1289 patients were recruited, and 117 patients were included for final analysis. On day of ICU admission, septic patients had significantly higher levels of serum HE4 than those with infection without sepsis, those with ovarian cancer, or healthy controls. Compared with septic survivors, septic non-survivors presented with significantly higher serum HE4 concentrations. Serum levels of HE4 correlated with disease severity scores and cytokine levels (IL-6 and IL-8). Upon ICU admission, the AUC for HE4 level association with 28-day mortality was 0.881, higher than the AUC for SOFA (0.713), IL-6 (0.589), and PCT (0.567). A regression analysis showed that HE4 was an independent mortality predictor. ConclusionHE4 can predict poor prognosis in septic patients, which may help to identify a group of septic patients at high risk of death.

NANOG regulates tumor metastasis via AMPK/mTOR signaling pathway in patients with epithelial ovarian cancer (252)

<b>Objectives:</b> NANOG engages with tumor initiation and metastasis by regulating EMT (epithelial-mesenchymal transition) in epithelial ovarian cancer (EOC). However, its role in association with pAMPK and its clinical significance in EOC has not been elucidated, even though pAMPK is known to degrade NANOG in various human cancer. Therefore, here we investigated the role of pAMPK and its association with NANOG as a potential prognostic, predictive biomarker in EOC. <b>Methods:</b> Immunohistochemistry of NANOG and pAMPK in epithelial ovarian cancer tissues were performed to define the clinical significance with TMA. Moreover, ovarian cancer cell lines were used for functional studies to investigate the underlying mechanisms. <b>Results:</b> Both NANOG and pAMPK expression were significantly overexpressed in EOCs compared to nonadjacent normal epithelial tissues, benign tissues, and borderline tumors. NANOG overexpression was significantly associated with poor DFS and OS, whereas pAMPK overexpression was associated with good DFS and OS. Importantly, multivariate analysis revealed that combining high NANOG and low pAMPK expression was an independent prognostic factor for DFS and associated with poor platinum resistance. In ovarian cancer cell lines, NANOG knockdown diminished migration and invasion property by regulating the EMT process via the AMPK/mTOR signaling pathway. Furthermore, treatment of AMPK activator suppressed expression of stemness-related genes, such as <i>NANOG, Oct4</i>, and <i>Sox2</i>. <b>Conclusions:</b> Collectively, our findings established that a combination of high NANOG and low pAMPK expression was associated with EOC progression and platinum resistance, suggesting a potential predictive biomarker for clinical management in EOC patients. Also, NANOG knockdown inhibited tumor metastasis and invasion by regulating EMT via the AMPK/mTOR signaling pathway.

Circulating ciRS-7 as a potential non-invasive biomarker for epithelial ovarian cancer: An investigative study.

BackgroundOvarian cancer (OC) is the most common malignant neoplasm of the female reproductive system in developed countries. Early detection, diagnosis and prognosis are particularly important to OC. The potential of circulating circular RNAs (circRNAs) as non-invasive biomarkers of various tumors has been especially described in recent years. The aim of this study was to explore the diagnostic and prognostic value of circulating cirRS-7 in patients with epithelial ovarian cancer (EOC).MethodsPre- and postoperative plasma samples from 111 EOC patients (47 cases with FIGO stage IA-IIB and 64 cases with FIGO stage IIB-IV) and healthy female volunteers was collected. Circulating ciRS-7 and hsa-miR-7-5p was analyzed using reverse transcription polymerase chain reaction (qRT-PCR). The diagnostic and prognostic value of circulating cirRS-7 as biomarker was estimated by the Receiver Operating Characteristic (ROC) curve and the area under the curve (AUC) and Kaplan–Meier analysis.ResultsThe preoperative expression levels of circulating ciRS-7 were increased in plasma of EOC FIGO stage I-IV patients than in the healthy controls (p < 0.001). However, the expression levels of ciRS-7 in the postoperative period were significantly lower in EOC FIGO stage IIA-IIA patients than healthy controls and EOC FIGO stage IIB-IV patients (p < 0.05, p < 0.001). The AUC of ciRS-7 for diagnosing EOC FIGO stage I-IV patients in pre-and postoperative periods was 0.90, 0.92, 0.84, 0.88, 0.58 and 0.86, respectively. Higher circulating ciRS-7 expression is associated with lymph node invasion, FIGO stage, distant metastasis, and worse overall survival (OS) of patients. Moreover, multivariate Cox analysis showed that higher circulating ciRS-7 was an independent predictor of OS in EOC FIGO stage IIB-IV patients. In addition, in plasma of EOC patients, ciRS-7 negatively correlated with has-miR-7-5p in pre-and postoperative periods (p < 0.001).ConclusionsCirculating ciRS-7 levels in plasma can be considered a potential candidate biomarker for diagnosing EOC patients. Dysregulation of ciRS-7 may participate in the molecular mechanism of EOC through hsa-miR-7-5p sponging.

Plasma exosomal miR-1260a, miR-7977 and miR-192-5p as diagnostic biomarkers in epithelial ovarian cancer.

Aim: The study aimed to clarify the diagnostic value of exosomal miRNAs in epithelial ovarian cancer (EOC). Methods: Plasma exosomes were isolated from peripheral blood of EOCpatients and healthy donors by ultracentrifugation and verified by transmission electron microscopy, qNano and western blot. The expression of exosomal miRNAs was detected by quantitativePCR, and the diagnostic efficiency of exosomal miRNAs was evaluated by receiver operating characteristic analysis. Results: Exosomal miR-1260a, miR-7977 and miR-192-5p were significantly decreased in EOC as compared with healthy controls. The area under the curve of the combination of three exosomal miRNAs was 0.8337. Moreover, the level of exosomal miR-7977 was related to the neutrophil-lymphocyte ratio, which decreased in EOC patients with ahigh neutrophil-lymphocyte ratio. Conclusion: Exosomal miR-1260a, miR-7977 and miR-192-5p act as potentially EOC diagnostic and prognostic biomarkers.

Abstract 5147: The prion-like protein doppel as a seurm biomarker for epithelial ovarian cancer

Abstract Ovarian cancer has a dire prognosis with an overall 5-year survival rate of ~40%, whereas stage III and IV has a survival rate of only 3-19%. Failure to detect early contributes highly to the poor survival rate of ovarian cancer. Thus, early detection of EOCs is likely to translate into mortality benefit. Two tests used clinically to detect ovarian cancer are serum cancer antigen-125 (CA-125) and transvaginal ultrasound. Both have limitations of specificity and sensitivity. CA-125 along with other biomarkers that are in clinical trials express both in the ovaries and serum of healthy females. Thus, the specific detection among cancer-free controls versus ovarian cancer cases remains highly variable. We hypothesize to assess the Doppel (PRND gene) as a serum biomarker in the detection of epithelial ovarian cancers (EOC) at the early stages. Doppel, a prion-like protein first identified in 1996, is known to express in testis and appears to regulate male fertility. Previously, we and others reported that Doppel is a highly specific tumor endothelium marker. In this study, to validate the potential of Doppel as an EOC serum biomarker, using ELISA, we detected Doppel concentration in the sera of 76 sera samples from patients with stage and histopathology information, and in 18 normal human sera (NHS). Doppel level was significantly higher in sera of EOC patients than NHS (0.720 vs 6.653 ng/ml, p&amp;lt;0.0001); 72 out of 76 patients were positive for Doppel expression (cut off value was calculated as Mean+3×SEM = 1.296 ng/mL) while only 3 out of 18 cases were positive for NHS. Receiver operating characteristic (ROC) analysis showed that Doppel can differentiate EOC patients from normal individuals with AUC of 0.957, sensitivity of 90.8%, and specificity of 90%. Doppel was markedly higher in all stages of EOC than NHS (Stage I, 6.261 ng/mL; Stage II, 8.045 ng/ml; Stage III, 7.040 ng/mL; and Stage IV, 5.726 ng/ml vs NHS 0.720 ng/ml; p&amp;lt;0.001). Interestingly, serum Doppel level was higher among EOC of serous adenocarcinoma (7.020 ng/mL; n = 39), mucinous (10.780 ng/mL; n = 8), and endometroid (7.295 ng/mL; n = 7) than clear cell (3.114 ng/mL; n = 7) or other types (4.855 ng/mL; n = 15). Together, our study showed the potential of Doppel as a serological biomarker for EOC detection. Citation Format: Xiaojun Zhang, Bowon Kang, Mazharul Karim, Jianying Zhang, Jae Yun Song, In-San Kim, Taslim A. Al-Hilal. The prion-like protein doppel as a seurm biomarker for epithelial ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5147.

Prognostic Impact of Circulating Methylated Homeobox A9 DNA in Patients Undergoing Treatment for Recurrent Ovarian Cancer.

Simple SummaryOvarian cancer remains a clinical challenge with considerable mortality. Circulating tumor DNA (ctDNA) has been suggested as a prognostic biomarker and enables the longitudinal evaluation of a patient’s disease and response to treatment. However, the role of ctDNA in treatment monitoring and for guiding treatment decisions in ovarian cancer remains unclear. We aimed to examine a gene methylation biomarker in the plasma of patients suffering a relapse of ovarian cancer in order to investigate prognostic potential and identify patients most likely to benefit from treatment, measured by overall survival. In the study, the methylated gene HOXA9 was found to be significantly related to poor survival, with the potential to observe the progression of the disease at an early stage and spare patients from ineffective treatment. Monitoring ctDNA during treatment is clinically feasible, further efforts are, however, required for standardization and for demonstrating improvement in treatment management.Methylated Homeobox A9 circulating tumor DNA (meth-HOXA9) has been suggested as a blood-based biomarker in epithelial ovarian cancer (EOC), although its prognostic significance remains unproven. The aim of the present study was to investigate the prognostic impact of meth-HOXA9 in patients with recurrent EOC. DNA was purified from 4 mL plasma and, following bilsulfite conversion, meth-HOXA9 was analyzed using a methylation-specific droplet digital PCR. Detection of meth-HOXA9 was reported as a percentage of total DNA and as a binary variable (detectable and undetectable). Meth-HOXA9 status and its dynamics during palliative treatment were correlated with overall survival (OS) as the primary endpoint. At baseline, meth-HOXA9 was detected in 65.9% (83/126) of the patients. The median OS was 8.9 and 17.9 months in patients with detectable and undetectable meth-HOXA9 at baseline (hazard ratio: 2.04, p = 0.002), which remained significant in the multivariate analysis. Median OS in patients with an increase in meth-HOXA9 after one treatment cycle was 5.3 months compared to 33 months in patients with undetectable meth-HOXA9 (p < 0.001). Meth-HOXA9 was significantly related to poor survival and may serve as a prognostic marker in patients with recurrent EOC. The longitudinal monitoring of meth-HOXA9 is clinically feasible with the perspective of aiding clinical decision making.