Diagnostic performance of the biomarkers HE4 and CA125 in type I and type II epithelial ovarian cancer

AbstractObjectiveTo evaluate the diagnostic performance of HE4 and CA125 in patients presenting with suspicious malignant ovarian cysts. We especially wanted to investigate the levels of HE4 and CA125 with regard to the gene and histology-unifying model of type I and type II epithelial ovarian cancer (EOC).MethodsPlasma from 373 women presenting with a suspicious malignant ovarian cyst was collected prior to surgery. Histology, grade, and stage were determined according to FIGO-classification. HE4 and CA125 were analyzed using ELISA, and the markers were evaluated for significance separately and in combination. Receiver operating curves, the area under the curve, sensitivity and specificity were estimated.ResultsThe combination of HE4 and CA125 resulted in the best diagnostic power in comparing benign tumors to EOC (ROC AUC 0.93, sensitivity 94.4% at 75% specificity) for type II. Diagnostic power in type I (ROC AUC 0.79, sensitivity 61.9% at 75% specificity) was less impressive. In particular, mucinous benign vs. malignant tumors could not significantly be separated by the dual marker combination. Impressively high ROC AUC 0.99 was found for the late stage type II EOC with 100% sensitivity at 75% specificity.ConclusionsHE4 and CA125 have a good ability to diagnose the more aggressive type II tumors but a poor diagnostic ability when patients are presenting with slow-growing type I in the early stage. Our results support the hypothesis that EOC should be looked upon as several different diseases, and that we lack biomarkers for sub-groups of EOC

External validation suggests Integrin beta 3 as prognostic biomarker in serous ovarian adenocarcinomas

<p>Abstract</p> <p>Background</p> <p>The majority of women with ovarian cancer are diagnosed in late stages, and the mortality rate is high. The use of biomarkers as prognostic factors may improve the treatment and clinical outcome of these patients. We performed an external validation of the potential biomarkers CLU, ITGB3, CAPG, and PRAME to determine if the expression levels are relevant to use as prognostic factors.</p> <p>Methods</p> <p>We analysed the gene expression of CLU, ITGB3, CAPG, and PRAME in 30 advanced staged serous adenocarcinomas with quantitative real-time polymerase chain reaction (QPCR) and the protein levels were analysed in 98 serous adenocarcinomas with western blot for semiquantitative analysis. Statistical differences in mRNA and protein expressions between tumours from survivors and tumours from deceased patients were evaluated using the Mann-Whitney U test.</p> <p>Results</p> <p>The gene and protein ITGB3 (Integrin beta 3) were significantly more expressed in tumours from survivors compared to tumours from deceased patients, which is in concordance with our previous results. However, no significant differences were detected for the other three genes or proteins CLU, CAPG, and PRAME.</p> <p>Conclusion</p> <p>The loss of ITGB3 expression in tumours from deceased patients and high expression in tumours from survivors could be used as a biomarker for patients with advanced serous tumours.</p

Potential predictive markers of chemotherapy resistance in stage III ovarian serous carcinomas

<p>Abstract</p> <p>Background</p> <p>Chemotherapy resistance remains a major obstacle in the treatment of women with ovarian cancer. Establishing predictive markers of chemoresponse would help to individualize therapy and improve survival of ovarian cancer patients. Chemotherapy resistance in ovarian cancer has been studied thoroughly and several non-overlapping single genes, gene profiles and copy number alterations have been suggested as potential markers. The objective of this study was to explore genetic alterations behind chemotherapy resistance in ovarian cancer with the ultimate aim to find potential predictive markers.</p> <p>Methods</p> <p>To create the best opportunities for identifying genetic alterations of importance for resistance, we selected a homogenous tumor material concerning histology, stage and chemotherapy. Using high-resolution whole genome array comparative genomic hybridization (CGH), we analyzed the tumor genomes of 40 fresh-frozen stage III ovarian serous carcinomas, all uniformly treated with combination therapy paclitaxel/carboplatin. Fisher's exact test was used to identify significant differences. Subsequently, we examined four genes in the significant regions (<it>EVI1</it>, <it>MDS1</it>, <it>SH3GL2</it>, <it>SH3KBP1</it>) plus the <it>ABCB1 </it>gene with quantitative real-time polymerase chain reaction (QPCR) to evaluate the impact of DNA alterations on the transcriptional level.</p> <p>Results</p> <p>We identified gain in 3q26.2, and losses in 6q11.2-12, 9p22.3, 9p22.2-22.1, 9p22.1-21.3, Xp22.2-22.12, Xp22.11-11.3, and Xp11.23-11.1 to be significantly associated with chemotherapy resistance. In the gene expression analysis, <it>EVI1 </it>expression differed between samples with gain versus without gain, exhibiting higher expression in the gain group.</p> <p>Conclusion</p> <p>In conclusion, we detected specific genetic alterations associated with resistance, of which some might be potential predictive markers of chemotherapy resistance in advanced ovarian serous carcinomas. Thus, further studies are required to validate these findings in an independent ovarian tumor series.</p

New Prognostic Markers in Stage III Serous Ovarian Adenocarcinomas

Ovarian carcinoma is the fifth most common cause of cancer death in Swedish women. Patients with advanced-stage disease respond differently to treatment, and the clinical outcome is difficult to predict for an individual patient. To increase the knowledge about ovarian adenocarcinomas, we aimed to investigate genetic changes relevant to the growth and progression of advanced ovarian tumours. We classified the tumours on a biological basis to identify potential biomarkers for use as prognostic factors. We analyzed the cytogenetic alterations with comparative genomic hybridization (CGH) and found significant differences in cytogenetic alterations in relation to survival, surgical outcome, and substage. Gain of regions at chromosome 1 and loss of regions at chromosome 4, 5, 8, 16, and X were common disparities associated with reduced survival. Gene expression analysis with microarray revealed a subgroup of survivors with a specific genetic signature that may be associated with less aggressive tumour progression or with tumours more sensitive to treatment. Quantitative real-time polymerase chain reaction (QPCR) was used to evaluate potential biomarkers and to narrow down the number of relevant genes to study at the protein level. Four genes, CLU, ITGB3, CAPG, and PRAME were differently expressed in tumours from survivors and tumours from deceased patients. We used western blot for semiquantitative analysis of the corresponding proteins, and found a significant difference in expression concerning survival for all four proteins. We performed an external validation of the four potential biomarkers in a new set of advanced ovarian adenocarcinomas. This established ITGB3 (Integrin beta 3) as significantly differently expressed concerning survival. The loss of ITGB3 expression in tumours from deceased patients and high expression in tumours from survivors could be used as a biomarker for patients with advanced serous tumours. In conclusion, we have found cytogenetic changes and differences in gene and protein expressions between advanced ovarian adenocarcinomas from survivors and deceased patients. These differences indicate that it is possible to predict the clinical outcome with a biological model for ovarian cancer patients

BAC CGH-array identified specific small-scale genomic imbalances in diploid DMBA-induced rat mammary tumors.

ABSTRACT: BACKGROUND: Development of breast cancer is a multistage process influenced by hormonal and environmental factors as well as by genetic background. The search for genes underlying this malignancy has recently been highly productive, but the etiology behind this complex disease is still not understood. In studies using animal cancer models, heterogeneity of the genetic background and environmental factors is reduced and thus analysis and identification of genetic aberrations in tumors may become easier. To identify chromosomal regions potentially involved in the initiation and progression of mammary cancer, in the present work we subjected a subset of experimental mammary tumors to cytogenetic and molecular genetic analysis. METHODS: Mammary tumors were induced with DMBA (7,12-dimethylbenz[a]anthrazene) in female rats from the susceptible SPRD-Cu3 strain and from crosses and backcrosses between this strain and the resistant WKY strain. We first produced a general overview of chromosomal aberrations in the tumors using conventional kartyotyping (G-banding) and Comparative Genome Hybridization (CGH) analyses. Particular chromosomal changes were then analyzed in more details using an in-house developed BAC (bacterial artificial chromosome) CGH-array platform. RESULTS: Tumors appeared to be diploid by conventional karyotyping, however several sub-microscopic chromosome gains or losses in the tumor material were identified by BAC CGH-array analysis. An oncogenetic tree analysis based on the BAC CGH-array data suggested gain of rat chromosome (RNO) band 12q11, loss of RNO5q32 or RNO6q21 as the earliest events in the development of these mammary tumors. CONCLUSIONS: Some of the identified changes appear to be more specific for DMBA-induced mammary tumors and some are similar to those previously reported in ACI rat model for estradiol-induced mammary tumors. The later group of changes is more interesting, since they may represent anomalies that involve genes with a critical role in mammary tumor development. Genetic changes identified in this work are at very small scales and thus may provide a more feasible basis for the identification of the target gene(s). Identification of the genes underlying these chromosome changes can provide new insights to the mechanisms of mammary carcinogenesis.JOURNAL ARTICLESCOPUS: ar.jinfo:eu-repo/semantics/publishe

Early Diagnosis of Epithelial Ovarian Cancer - Analysis of Novel Biomarkers

Early Diagnosis of Epithelial Ovarian Cancer - Analysis of Novel Biomarkers Björg Kristjánsdóttir Department of Obstetrics & Gynecology, Institute of Clinical Sciences at Sahlgrenska Academy, University of Gothenburg, Sweden ABSTRACT Background: Majority of epithelial ovarian cancer (EOC) is detected in advanced stage with bad prognosis and high mortality. Reliable diagnostic markers are lacking, pre-cancerous lesions in the more aggressive tumors are not clearly defined, vague or unspecific early symptoms, and the localization of the ovaries, deep in the pelvis contributes to late diagnose. Heterogeneity, not only different type of histology, but also different intrinsic biology and behavior characterizes ovarian cancer. Invasive surgery with histological examination is needed to confirm the diagnosis. Less than 25% EOC are diagnosed early, when there is great possibility to cure and 5-year survival >90%, in contrast to 20-30% 5-year survival in late stage EOC. Thus, early detection is of utmost importance. Proximal fluids, like ovarian cyst fluid, are promising in the search for early markers. Cancer antigen 125 (CA125), the most used biomarker since 30 years, and a promising marker human epididymis 4 (HE4) have recently been approved by FDA to be used in the prediction of malignancy in women with a pelvic mass. Aims: To explore ovarian cyst fluid as a source mining for new diagnostic biomarkers for EOC, and to validate the markers found together with CA125 (Paper I-III); and to evaluate the diagnostic performance of HE4 and CA125, to distinguish between benign cysts and EOC, and EOC divided into slow growing type I and the aggressive type II EOC (Paper IV-V). Method: Cross sectional, observational, explorative, and diagnostic clinical studies, with prospective and consecutive collection of cystic fluid, blood and tumor tissue at the time of operation and retrospective analysis. Women with suspicious malignant pelvic cysts, already scheduled for operation at our clinic for tumor surgery were included. High throughput proteomic analyses were used for searching for novel markers, and selected proteins were validated with ELISA or immunoblot. Paper I: The cyst fluid proteome was mined with surfaceenhanced laser desorption/ionization time of flight (SELDI-TOF) mass spectrometry (MS) (n=192). Paper II: Enrichment of a selection of known cancer antigens to overcome high abundant proteins, and with focus on inflammation, was followed by Immunoprecipitation MS (n=38). Significantly differently expressed chemokines were validated (n=256). Paper III: Serous cystadenoma (n=5) and serous adenocarcinoma (n=10) of different stages were analyzed with isobaric tag for relative and absolute quantification (iTRAQ), followed by immunoblot validation (n=68). Paper IV-V: HE4 and CA125 levels in plasma were analyzed with ELISA and Risk of Ovarian Malignancy Algorithm (ROMA) was calculated (n=393). Significant differences, receiver operator characteristics (ROC) area under the curve (AUC), cut-off levels, sensitivity and specificity were estimated with regard to malignancy, grade, stage histologic subtype and type I and type II. Results: Paper I: Combination of Apolipoprotein CIII and Protein C inhibitor had the best AUC (0.91) in cyst fluid, and improved by CA125 (0.94). Abundant proteins were a problem in the cyst fluid analyses. Paper II: Interleukin-8 and Chemoattractant Protein-I were highly significantly increased expressed in cyst fluid. Increased inflammatory response was present in early tumor development and earlier than in blood. Paper III: Two of 87 differentially expressed proteins in cyst fluid, with high significance and fold change, Serum Amyloid A-4 (SAA4) and astacin-like metalloendopeptidase (ASTL) were validated, and SAA4 was significantly increased in cyst fluid, but not in blood. Paper IV: HE4 complemented CA125 in the diagnosis of ovarian cysts, especially in the premenopausal women. Sensitivity for ROMA at set specificity of 75% was highest in the postmenopausal cohort (87%). Paper V: HE4 and CA125 diagnosed the aggressive type II EOC most correctly (AUC 0.93), but the results were not acceptable in early stage type II (AUC 0.85) or in type I EOC (AUC 0.79) respective early type I AUC 0.73). Conclusion: Ovarian cyst fluid is an excellent source for the search of novel biomarkers for early diagnosis of EOC. Early events are found near the tumor in the early phase, like the inflammatory response and later on in the peripheral circulation. HE4 complements CA125 in predicting malignancy in cystic ovarian tumors. The result from this thesis support, that EOC should be looked upon as several different diseases. Finding early markers that are specific for each histology subgroup will be the future challenge. Combination of such markers in a panel could improve the early diagnosis of EOC. Keywords: EOC; ovarian adenocarcinoma; ovarian cyst fluid; pelvic mass; tumor biomarker; mass spectrometry; SELDI-TOF MS; iTRAQ; ISBN 978-91-628-8727-8 http://hdl.handle.net/2077/3309