Dissertations / Theses: 'High-grade serous ovarian carcinoma'

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Everitt, Gemma Louise Ann. "The inflammatory infiltrate of high-grade serous carcinoma omental metastasis." Thesis, Queen Mary, University of London, 2014. http://qmro.qmul.ac.uk/xmlui/handle/123456789/8038.

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The aim of this thesis is to investigate the role of inflammatory infiltrates and chemokines in metastasis of high-grade serous ovarian cancer, HGSC, to the omentum using human tissue biopsies and a 3- dimensional (3D) cell culture model. In ten patients with metastatic HGSC, omental tumour deposits contained a prominent leukocyte infiltrate of CD3+ T cells (9% of total cells) and CD68+ macrophages (11% of total cells). The presence of CD68+ macrophages showed a significant positive correlation with tumour cell proliferation analysed by Ki67 expression. Four ovarian cancer cell lines were co-cultured on a 3D model mimicking the microenvironment of the omentum for two weeks. The model was composed of collagen embedded human fibroblasts covered in a confluent layer of human primary mesothelial cells. The mesothelial cells in the 3D model significantly increased the growth (p = 0.002) and invasion (p = 0.0004) of the ovarian cancer cells. CXCL12 is the macrophage chemoattractant and ligand for the major chemokine receptor expressed on ovarian cancer cells. An association between CXCL12 and extracellular matrix remodelling was identified in two independent gene expression microarrays of ovarian cancer biopsies. The expression of CXCL12 in the HGSC omental metastases measured by quantitative Real Time-PCR positively correlated with decorin expression. Antibody mediated neutralisation of CXCL12 reduced growth (p = 0.012) and invasion (p = 0.029) in the 3D model. Mimicking an infiltrate of CD68+ macrophages in this multicellular 3D in vitro system also produced measurable changes in inflammatory cytokine and chemokine expression. There is currently a demand for more accurate models of HGSC and a necessity to study its metastasis that presents itself as the major clinical problem in patients. Therefore the development of this 3D model to mimic tumour-promoting inflammation in HGSC metastasis will provide researchers with an essential tool for testing novel therapeutic strategies.

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Sheets, Jordan N. "Inhibitory Functions of SUSD2 in the Progression of High-Grade Serous Ovarian Carcinoma." Thesis, University of South Dakota, 2017. http://pqdtopen.proquest.com/#viewpdf?dispub=10633628.

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Sushi Domain Containing 2 (SUSD2) encodes a type I transmembrane protein containing several functional domains inherent to adhesion molecules. A clinically annotated HGSOC tissue microarray was stained with an anti-SUSD2 antibody. Patients with tumors that had weak SUSD2 staining had a shorter median survival (31.7 months) compared to patients that had tumors with strong SUSD2 staining (49.1 months; p value = 0.0083).

To investigate the role of SUSD2 in HGSOC, stable OVCAR3, OVSAHO and KURAMOCHI cell lines were established through transfection of shRNA targeted to SUSD2 transcripts (SUSD2 knock-down [KD] cell lines) or non-targeting shRNA (SUSD2 NT) cell lines. Boyden chamber and wound healing assays demonstrated that OVCAR3, OVSAHO and KURAMOCHI SUSD2 KD cells migrated at significantly higher rates than their SUSD2 NT counterpart cell lines. RT-qPCR and western immunoblot analysis indicated an inverse relationship between SUSD2 and well characterized mesenchymal genes, such as TWIST1, ZEB1 and CHD2. In addition, OVCAR3 and KURAMOCHI SUSD2 KD spheroids displayed increased mesothelial clearance ability compared to SUSD2 NT spheroids.

To explore the potential for SUSD2 to inhibit late-stage HGSOC metastasis, female athymic nude mice were injected with either OVCAR3 NT or OVCAR3 KD cells. Fewer nodules were observed in the pancreas and omentum of the OVCAR3 NT mice when compared to the OVCAR3 KD mice. Furthermore, OVCAR3 KD mice had a significantly shorter median survival compared to OVCAR3 NT mice (175 days compared to 185.5 days, respectively; p-value = 0.0047).

KURAMOCHI lysate was immunoprecipitated for SUSD2-associated immunocomplexes and subjected to liquid chromatography, tandem mass spectrometry (LC-MS/MS) analysis, yielding a list of candidate SUSD2-interacting proteins associated with RNA processing. Immunofluorescence analysis of OVCAR3, KURAMOCHI and SKBR3 cells and western immunoblot analysis of their subcellular extracts revealed SUSD2 to be present in cell nuclei, mitochondria and cytoplasm; however, SUSD2 was relatively less abundant in SKBR3 nuclei.

Our findings suggest that increased SUSD2 expression in HGSOC impedes metastasis, consistent with prolonged survival observed in HGSOC patients with high SUSD2-expressing primary tumors. The differences in subcellular distribution between HGSOC cells and breast cancer cells may explain alternate functions of SUSD2 in different cancers.

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Cole, Alexander John. "The role of p53 and histone H2B monoubiquitination in high-grade serous ovarian carcinoma." Thesis, The University of Sydney, 2017. http://hdl.handle.net/2123/17639.

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Ovarian cancer is the eleventh most frequently diagnosed cancer in women and the fifth most common cause of cancer-related deaths. Epithelial ovarian cancer accounts for ~90% of cases. Serous epithelial ovarian cancer (SEOC) is the most common and aggressive histological subtype. Clinically, SEOC can be divided into type I and II tumours. Type II tumours (high grade serous ovarian carcinoma (HGSOC)), are generally diagnosed at later stages, grow aggressively and possess a mutation in the TP53 gene, which codes for the tumour suppressor protein p53. Mortality rates of HGSOC have remained largely unchanged for decades due to a lack of sensitive biomarkers and poor response to primary treatment. Finding new strategies to treat patients and identify early disease is critical to improving patient outcomes. Post-translational histone modifications describe the chemical and protein alteration of histone tail residues which are capable of altering gene expression. Monoubiquitination of lysine 120 on histone H2B (H2Bub1) disrupts chromatin strands and has important physiological functions including transcription, DNA repair, differentiation and crosstalk. Recent research has demonstrated that H2Bub1 is lost in cancer suggesting it may play an important role in malignancy. Interestingly, several studies have reported a link between the tumour suppressor p53 and H2Bub1, including a direct interaction of p53 with the H2Bub1 E3 ligase RNF20, and H2Bub1 enrichment at the p53 target gene CDKN1A. Consequently, this link may prove important in HGSOC where TP53 mutations occur almost ubiquitously and result in the loss of wildtype p53 function. 2 At the commencement of this work no study had comprehensively investigated the relationship between TP53 mutation status and p53 immunohistochemistry (IHC) using massively parallel sequencing (MPS) in HGSOC. The aim of this work was to determine the accuracy of p53 IHC to determine TP53 mutation status. We used MPS and IHC to characterise HGSOCs for TP53 mutation and p53 expression. Mutations in TP53 were identified in 94% (68/72) of HGSOCs, 62% of which were missense. Missense mutations demonstrated high p53 staining by IHC, as did 35% (9/26) of non-missense mutations. Low p53 staining was seen by IHC in 62% of HGSOC associated with non-missense mutations. Most wildtype TP53 tumours (75%, 6/8) displayed intermediate p53 expression levels. The overall sensitivity of detecting a TP53 mutation based on classification as ‘Low’, ‘Intermediate’ or ‘High’ for p53 IHC was 99%, with a specificity of 75%. We suggest p53 IHC can be used as a surrogate marker of TP53 mutation in HGSOC; however, this will result in misclassification of a small proportion of TP53 wildtype and mutant tumours. H2B monoubiquitination can be catalysed by five E3 ligases, and removed by up to eleven deubiquitinases. Loss of H2Bub1 has been reported in a range of malignancies, where it associates with higher tumour stage/grade, metastasis and poor patient survival. The aim of this section of work was to investigate H2Bub1 levels in HGSOC and correlate them with p53 mutation status/IHC, the deubiquitinase USP7 and the polycomb repressive complex subunit enhancer of zeste homolog 2 (EZH2), which is capable of negatively regulating H2Bub1 levels by competing for the same substrate. H2Bub1 was lost in 68% of HGSOC samples, but did not correlate with overall patient survival. P53 intermediate IHC and wildtype TP53 status correlated with higher H2Bub1 IHC staining, while USP7 expression was inversely correlated with H2Bub1 IHC. EZH2 expression did not correlate with H2Bub1 IHC or with patient overall survival. In vitro experiments using wildtype and mutant p53 3 constructs transfected into p53 functionally null (herein referred to as null) cell lines appeared to contradict the p53 and H2Bub1 IHC data, warranting further investigation. Along with other studies, our data suggested a link between p53 and H2Bub1. Consequently, this work aimed to investigate the effects of p53 on H2Bubl levels. We treated p53 wildtype cell lines with cisplatin, which activated p53 and resulted in global loss of H2Bub1 levels. In contrast, treatment of p53 null cell lines did not significantly alter H2Bub1 levels. Transfection of wildtype p53 into the same null cell lines resulted in a significant decrease in H2Bub1 levels compared to the vector only control. Downregulation of the main H2Bub1 E3 ligase, RNF20, and treatment with cisplatin showed a decrease of p53 levels in RNF20 sRNA treated cells compared to control siRNA. Chromatin immunoprecipitation coupled with next generation sequencing (ChIP-seq) of a p53 wildtype cell line treated with cisplatin, demonstrated H2Bub1 enrichment at p53 target genes. RNA-seq demonstrated an increased expression of these H2Bub1 enriched p53 target genes. Downregulation of RNF20 using siRNA, sensitised MCF7 and A2780 cells to cisplatin treatment. Together, this work suggested a functional link between H2Bub1 and p53. Chemoresistance is a hallmark of HGSOC and has been linked to the presence of cancer stem cells (CSCs). Ovarian CSCs make up a small subpopulation of cells within the primary tumour mass and express the markers CD133 and ALDH. Recent reports have suggested H2Bub1 plays an important role in stem cell differentiation and maintenance. The aim of this section was to investigate the role of H2Bub1 in ovarian CSCs. H2Bub1 was shown to be depleted in CD133+ cells and spheroids enriched for ALDH, consistent with a more stem-like phenotype. Decreasing H2Bub1 levels through RNF20 downregulation resulted in a profound effect on the CSC populations, where ALDH+ populations increased and the CD133+ 4 populations decreased. Spheroid culture of the RNF20 downregulated cells resulted in larger and more numerous spheroids than the shRNA control suggesting RNF20 and hence H2Bub1, can influence spheroid formation. Together, this data suggests H2Bub1 may play an important role in CSC differentiation.

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Cheng, Jung-Chien. "Role of E-cadherin in the serous borderline ovarian tumor and low-grade serous ovarian carcinoma cell invasion." Thesis, University of British Columbia, 2012. http://hdl.handle.net/2429/43651.

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E-cadherin is a membrane glycoprotein located at cell adherens junctions. A switch from E-cadherin to N-cadherin expression has been considered a hallmark of the epithelial-mesenchymal transition (EMT), which is primarily due to the up-regulation of the transcription factors Snail, Slug, Twist and ZEB1. Epithelial ovarian cancer cells with low E-cadherin expression are more invasive, and the absence of E-cadherin expression in ovarian cancer is associated with poor prognosis and survival. Serous borderline ovarian tumors (SBOT) are slow-growing, non-invasive ovarian epithelial neoplasms. SBOT are considered distinct entities that give rise to invasive low-grade serous carcinomas (LGSC), which have a relatively poor prognosis and are unrelated to high-grade serous carcinomas (HGSC). The mechanisms underlying the progression of non-invasive SBOT to invasive LGSC are not understood. We have established short-term cultures of SBOT cells from tumor biopsies and have shown that inactivation of p53, Rb and/or PP2A by the SV40 large T (LT) and small T (ST) antigens allows SBOT cells to acquire characteristics associated with neoplastic progression, including increased cell motility, invasion and EMT. However, the overexpression of N-cadherin does not induce cell invasion in SBOT cells. In this study, using loss- and gain-of-function approaches, we show that p53 acts as a tumor suppressor in the regulation of SBOT and LGSC cell invasion by regulating E-cadherin expression through PI3K/Akt-mediated transcriptional and epigenetic machineries. In high-grade ovarian cancer cultures, it has been shown that epidermal growth factor (EGF) and transforming growth factor-beta (TGF-β) induce cell invasion by activating the EMT. However, the effects of EGF and TGF-β on SBOT and LGSC cell invasion remain unknown. We show that EGF induces SBOT cell invasion by activating the EMT. In addition, our results suggest that there are EMT-independent mechanisms that mediate EGF-induced LGSC cell invasion. Interestingly, we show a dual function for TGF-β in which it induces invasion in SBOT cells by activating the EMT and promotes apoptosis in LGSC cells. Overall, this study demonstrates that the loss of E-cadherin expression in SBOT may play an important role in the transition to invasive LGSC.

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Virtanen, Siru Sirkku Pauliina. "Phenotypic and functional characterisation of cancer stem cells in human high-grade serous ovarian carcinoma." Thesis, University of Cambridge, 2014. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.648879.

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6

Murakami, Ryusuke. "Establishment of a novel histopathological classification of high-grade serous ovarian carcinoma correlated with prognostically distinct gene expression subtypes." Kyoto University, 2016. http://hdl.handle.net/2433/215449.

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Dawson, Amy. "Targeted therapy in low-grade serous ovarian carcinoma : characterization of MEK inhibitor response in novel patient-derived cell lines." Thesis, University of British Columbia, 2016. http://hdl.handle.net/2429/59456.

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Low-grade serous ovarian carcinoma, or LGSOC, predominantly occurs in pre- menopausal women at lower frequency than the more common high-grade serous ovarian carcinoma. It tends to harbour few, but distinct, mutations, with a significant proportion of RAS oncogene mutations. Aberrations of the RAS-MAPK signaling pathway in LGSOC have led to the initiation of several clinical trials of MEK inhibitors in this disease. Due to the relative rarity of LGSOC, few representative model systems are available. Accordingly, there has been a lack of preclinical investigational drug testing in this disease. In this study, 11 histotype-specific cell lines have been derived and molecularly characterized in order to further elucidate the molecular biology of the disease, and serve as an important platform to test novel therapeutics in LGSOC. The most common missense mutations by HotSpot mutational profiling analysis were oncogenic KRAS/NRAS mutations, and varying levels of copy number aberration were seen. All cell lines were uniquely identifiable by short-tandem repeat analysis, with the exception of two patient-paired cell lines. The phenotypic and molecular responses to the 4 MEK inhibitors (MEKi) trametinib, selumetinib, refametinib, and binimetinib have been characterized in selected LGSOC cell lines through IC50 analysis, proliferation, viability, and apoptosis assays, and on-target drug effects by Western blot. Of the 4 MEKi, trametinib exhibited the best anti-proliferative effects and inhibition of MEK kinase activity. Biological and on-target data from two of the cell lines reveals an exquisite sensitivity to MEK inhibition. Reverse-phase protein array and quantitative mass spectrometry global proteomic analysis on control and MEKi-treated LGSOC cell lines was performed in order to examine basal proteomic differences between MEKi-sensitive and resistant LGSOC cell lines and gather data for examining proteomic changes upon MEKi treatment. Three significantly differentially expressed protein candidates (PKCα, EGFR, and Smac/DIABLO were identified between sensitive and resistant cell lines under control and MEKi treatment conditions by both proteomic platforms. Results from therapeutic testing in these pathologically reviewed, histotype- specific LGSOC models allow for a comparison of the overall efficacies of the 4 MEKi, characterization of drug sensitivity in novel cell lines, and identification of potential functional markers of MEKi response.
Medicine, Faculty of
Graduate

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Howe, Eleanor Arden. "MicroRNA expression and activity in high-grade serous ovarian cancer." Thesis, University of Oxford, 2012. http://ora.ox.ac.uk/objects/uuid:9d17590c-550b-4ae9-ac8d-15387cf70e5f.

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miRNAs are critical modulators in the development and progression of cancer. Emerging evidence suggests that they are drivers of ovarian cancer. A better understanding of the molecular underpinnings of the development, progression and chemoresistance of the disease is critical for the development of new, more eﬀective therapies. Here we explore the expression patterns of miRNAs as they relate to gene expression, as they diﬀer across molecular subtypes of the disease. We examine the correlation structure of miRNA expression with mRNA expression in two distinct genomic datasets and report on patterns in correlation structure in several subsets of the data. We ﬁnd that the datasets show consistency in their correlation structure, and in the speciﬁc miRNA-mRNA pairs that are either highly positively or negatively correlated. The data include a larger number of strong positive and strong negative correlations than would be expected by chance, indicating that biological relationships between the types of data are detectable in these datasets. We further ﬁnd an enrichment for positively-correlated miRNA-mRNA pairs in which the miRNA is encoded in close proximity to the mRNA. The correlation of miRNA and mRNA is apparently unaﬀected by miRNA and mRNA expression level; similarly the two molecular subtypes do not contain diﬀerences in their correlation. We ﬁnd that the recently described poorer prognosis, or angiogenic, subtype has a generally lower miRNA activity than the second, non-angiogenic, subtype. The subtypes are characterized by a consistent pattern of diﬀerential miRNA expression. We also report on a switch-like relationship between the expression levels of certain miRNAs and the genes that are anticorrelated with them. We propose these miRNAs drive many of the diﬀerences in the subtypes both directly, by RISC-mediated repression of target messages and indirectly, by repressing transcription factors that regulate expression in the cell. We build models of patient survival and time-to-relapse based on these miRNA expression data and inferred miRNA activity scores, using several types of univariate and variable selection models. We ﬁnd essentially no survival-predictive information provided by the RE score data. While the direct miRNA expression measurements may contain some predictive power, we ﬁnd that a larger dataset and the segretation of that dataset into distinct molecular phenotypes is likely to be necessary to produce a useful model of survival in ovarian cancer.

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Todeschini, P. "NON-CODING RNAS IN HIGH-GRADE SEROUS EPITHELIAL OVARIAN CANCER." Doctoral thesis, Università degli Studi di Milano, 2016. http://hdl.handle.net/2434/488108.

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Abstract Introduction: High-grade serous ovarian carcinoma (HGSOC) is the most lethal gynecologic malignancy, mainly because the disease is frequently diagnosed at an advanced stage and is characterized by the early onset of chemoresistant recurrences. The lack of reliable diagnostic and prognostic markers, together with the lack of effective therapies, are the major obstacles to the clinical management of patients with HGSOC. A new class of non-coding RNAs (ncRNAs), such as microRNA (miRNAs) and long non-coding RNAs (LncRNAs), with a function of gene expression regulation, have been discovered to play an important role in human cancers. Increasing evidences suggest that ncRNAs are involved in cancer progression and development of chemoresistance, and support their role as potential diagnostic, predictive and prognostic biomarkers. The hypoxic condition within the tumor microenvironment, improving the tumor neovascularization, represents an essential event contributing to the development of a more aggressive HGSOC phenotype. Recently, a group of miRNAs, termed hypoxia regulated-miRNAs (HRMs), have been identified as key elements in response to hypoxia, regulating important mechanisms involved in tumor progression. The complexity of hypoxia molecular mechanisms has not been fully elucidated yet in HGSOC, therefore there is an urgent need to discover novel biomarkers clinically useful to select patients with hypoxic tumor, that may benefit of tailored treatments. Aims of the study: My PhD project aims at elucidating transcriptional and post-transcriptional signatures characterizing HGSOC, both at the serum and tissue levels. In detail, the research effort includes: i) the investigation of circulating miRNAs as novel potential biomarkers for HGSOC detection; ii) the analysis of mRNA, miRNA and lncRNA expression profiles of HGSOC and normal tissues; iii) the evaluation of hypoxia-regulated miRNA expression in HGSOC and normal tissues. Methods: Sera from 168 HGSOC stage III-IV patients and 65 healthy donors were gathered together from two independent collections and stratified into a training set, for miRNA marker identification, and a validation set, for data validation. Nine synthetic viral/C.Elegans spike-in oligos were added to serum samples before RNA extraction, to allow accurate normalization. miRNA expression profiles were obtained using Agilent Microarray Technologies®. An innovative statistical approach for microarray data normalization, based on the contribute of spike-in oligos and the most invariant miRNAs, was developed to identify, in the training set, differentially expressed miRNAs. Signature validation in both the training and validation sets was performed by Real Time quantitative PCR (RT-qPCR) and confirmed by droplet digital PCR (ddPCR). A total of 99 tumor biopsies were collected from HGSOC stage III-IV patients, partially matched with the serum sample cohort (n=76). Thirty normal tissues were obtained from normal ovary (HOSE) and luminal fallopian tube surface epithelia, both representing the normal counterpart for HGSOC, whose histogenesis is still a matter of debate. Gene and miRNA expression profiles were obtained using Agilent Microarray Technologies®. miRNA expression levels were correlated with patient outcomes, as overall survival (OS) and progression-free survival (PFS). Additionally, a subgroup of 14 chemo-resistant and 14 chemo-sensitive HGSOC patients, together with 10 normal tissues were deep sequenced for the discovery of novel HGSOC specific coding and non-coding transcripts. Results: A panel of 97 miRNAs emerged significantly differentially expressed (92 up-regulated and five down-regulated) between sera of HGSOC patients and healthy donors by microarray analysis. Among them, the following miRNAs, i.e., miR-1246, miR-595, miR-574-5p, miR-483-3p, miR-4290, miR-2278, miR-32, miR-4281, and miR-3148, exhibiting both the highest average expression and log fold change measured in patients compared to healthy donors, were selected for further validation. miR-1246, miR-595 and miR-2278 were confirmed as significantly over-expressed in serum of HGSOC patients compared to controls by RT-qPCR (all p-values<0.03), in both the training and validation sets. Receiver Operating Characteristic (ROC) curve analysis revealed miR-1246 as the best diagnostic biomarker, with a sensitivity of 87%, a specificity of 77% and an accuracy of 84%. The absolute quantification of circulating miR-1246 by ddPCR confirmed its potential as diagnostic biomarker in HGSOC. Microarray analysis of tissue miRNA profiling revealed a total of 265 miRNAs significantly dysregulated (123 up-regulated and 142 down-regulated) in HGSOC compared to normal tissues. A group of nine miRNAs (i.e., miR-199b-5p, miR-423-5p miR-455-3p, miR-22-3p, miR-199a-3p, miR-15b-5p, miR-140-5p, miR-1246, and miR-320c) were associated with platinum response and prognosis. In particular, among tumor samples, miR-1246 up-regulation was consistently associated with platinum-resistance, poor OS and poor PFS (p-values<0.05). Kaplan-Meier survival curves, according to miR-1246 expression levels obtained by RT-qPCR, showed that OS and PFS decreased in patients with high miR-1246 expression compared to those with low miR-1246 expression (p-value<0.001, HR=2.57; p-value=0.024, HR=1.68; respectively). In addition, multivariate analysis revealed miR-1246 over-expression as an independent prognostic factor for poor OS and PFS (p-value=0.002, HR=2.31; p-value<0.05, HR=1.59; respectively). Interestingly, compared to normal tissues, both with microarray and RT-qPCR techniques, miR-1246 showed a down-regulation compared to HOSEs (p-value<0.0001), but we did not detect a significantly differential expression compared to fallopian tubes. This result mirrors the global miRNA expression trend revealed by principal component analysis (PCA) on microarray data. Subsequently, we focused our analysis on a group of 16 miRNAs belonging to the group of hypoxia-regulated miRNAs (HRMs) emerged from literature as relevant in other solid tumors. Among them, we confirmed miR-210 and miR-27a-3p/23a-3p/24-3p cluster as significantly up-regulated in HGSOC vs normal tissues by RT-qPCR (all p-values≤0.002). More interestingly, we validated the significant over-expression of miR-23a-3p in the group of patients resistant to platinum-based chemotherapy compared to platinum-sensitive patients (p-value=0.03). In addition, in univariate survival analysis miR-23a-3p over-expression showed a significant correlation with decreased progression-free survival (p-value=0.009, HR=1.8), but not with overall survival variable. Importantly, miR-23a-3p over-expression has emerged as an independent prognostic marker for shortened progression-free survival in multivariate Cox regression analysis (p-value=0.01, HR=1.78). Finally, the preliminary analysis of the transcriptome sequencing allowed us to identify 1371 transcripts differentially expressed between platinum-resistant and platinum-sensitive samples. Among them, 125 transcripts showed a complete match of intron chain with known transcripts, 686 were potentially novel isoforms or showed a generic overlap with known transcripts. The remaining 560 sequences, if validated, could be novel intergenic transcripts or transcripts with an exonic overlap with reference ones. Conclusions: This study demonstrates, for the first time, miR-1246 as a potential diagnostic serum biomarker in HGSOC, as assessed by three independent technologies (microarray, RT-qPCR and ddPCR) and validated in two independent cohorts of patients. Moreover, high-throughput analysis reveals most of the gene and miRNA dysregulated in HGSOC biopsies compared to the normal counterpart. In particular, our findings indicate, for the first time, that miR-1246 over-expression correlates with a platinum-resistant HGSOC phenotype and may constitute a novel prognostic factor for HGSOC patients. Furthermore, our results regarding HRMs suggest an important role of miRNAs in response to hypoxic conditions within HGSOC. Particularly, the miR-23a-3p over-expression in the group of platinum-resistance patients may contribute to explain the importance of hypoxia in HGSOC mechanism of drug resistance and could represent an independent prognostic marker for HGSOC patients. Lastly, preliminary data emerged from transcriptome analyses, suggesting a prominent non-coding role in HGSOC platinum resistance, will be integrated with gene and miRNA expression profiles previously obtained, with the aim to identify tumor circuits associated with response to treatment and prognosis, as well as to better elucidate the molecular mechanisms characterizing HGSOC progression and adaptation to hypoxic tumor microenvironment.

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Moore, Elizabeth. "Improving earlier non-invasive diagnosis of high-grade serous ovarian cancer." Thesis, University of Cambridge, 2018. https://www.repository.cam.ac.uk/handle/1810/285405.

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The majority of women with ovarian cancer (OC) have advanced disease at diagnosis and 5-year survival rates of less than 25%. Women with stage I disease have significantly better 5-year survival rates of over 90%. Recent large studies using CA 125 and transvaginal ultrasound have failed to improve mortality in a screened population. There is therefore a pressing need for new diagnostic biomarkers in OC. The primary aim of my project, as a first step in developing a diagnostic circulating tumour DNA (ctDNA) biomarker for high grade serous ovarian cancer (HGSOC), was to investigate low-cost high-throughput next generation sequencing assays in plasma samples collected from women with newly diagnosed OC. The secondary aim was to apply these methods to other non-invasive samples including cervical liquid based cytology samples that might contribute to earlier diagnosis or screening for women with OC. ctDNA was detected in 30-49% of women with newly diagnosed OC from the UKOPS (n=54) and CTCR-OV04 (n=156) cohorts using targeted sequencing. Using the trimmed median absolute deviation (t-MAD) score, a quantitative measure of genome wide copy number aberration generated from shallow whole genome sequencing (sWGS) data, ctDNA was detected in 39-41% of the women with newly diagnosed disease. To improve sensitivity of ctDNA detection I developed an optimised method for targeted sequencing that has the potential to lower the limit of detection of ctDNA in HGSOC by 100 fold. I have also shown that the size profile of HGSOC ctDNA fragments is different to that of wildtype DNA fragments and shown that selecting for DNA fragments between 90-150 bp can increase rates of ctDNA detection in HGSOC. ctDNA detection increased to 53-67% of women with newly diagnosed OC using the size selected t-MAD score. I have evaluated the utility of cervical sampling for earlier diagnosis of OC by testing and optimising DNA extraction, library preparation and sequencing methods. I have detected tumour DNA in routine cervical cytology samples collected from women subsequently diagnosed with cervical and endometrial cancers. In summary I have developed methods for ctDNA detection in women with newly diagnosed HGSOC that can be applied and refined in larger prospective studies of women undergoing follow-up for treated HGSOC, women with symptoms suggestive of OC and women at high risk of OC.

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Shahabi, Shohreh. "Exploring Novel Precision Medicine Approaches in High Grade Serous Ovarian Cancer." Doctoral thesis, Universite Libre de Bruxelles, 2020. https://dipot.ulb.ac.be/dspace/bitstream/2013/312182/3/ToC.pdf.

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In this dissertation, we aimed to bring together a team of clinical experts, translational researchers, biostaticians and bioinformaticians to develop and implement innovative scientific methodologies in precision medicine applied to High Grade Serous Ovarian Cancer (HGS OvCa). We used a variety of translational and computational methods in order to generate impactful outcomes. These pipelines produced statistically robust results, with particular emphasis on drawing clinical and biological correlations. The results presented here contribute to the body of evidence necessary to substantiate these findings in a clinical setting. Bioassays, PDX models and ancillary specimen evaluation of previous clinical trials will help to validate our candidate biomarkers. Enhanced understanding of the molecular pathology of disease grounded in acquisition of genomic knowledge will facilitate the development of targeted treatment in cancer. Because clinical trials must be developed with correct metrics, patient selection and drug efficacy should incorporate adaptive designs.
Doctorat en Sciences médicales (Santé Publique)
info:eu-repo/semantics/nonPublished

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OPINATO, VIVIANA MARIA. "Role of Mesenchymal Stromal Cells in High-Grade Serous Ovarian Cancer." Doctoral thesis, Università degli Studi di Trieste, 2018. http://hdl.handle.net/11368/2924766.

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Mesenchymal Stromal Cells (MSCs) are the precursors of various cell types that compose both normal and cancer tissue microenvironments. In order to support the widely diversified parenchymal cells and tissue organization, MSCs are characterized by a large degree of heterogeneity, although available analysis of molecular and transcriptional data do not provide any clear evidence. Moreover, a wealth of study has demonstrated a significant role of the microenvironment and MSCs in tumor growth. In the laboratory in which I carried out my PhD project, MSCs were purified from different healthy tissues (N-MSCs) and from High-Grade Serous Ovarian Carcinomas (HG-SOC-MSCs). In order to study the role of MSCs in High-Grade Serous Ovarian Cancer, two-dimensional (2D) simplified co-culture approaches were established to characterize the interaction between MSCs and an ovarian carcinoma cell line – Skov3 – at short-term (1 hour) and long-term (5 days) settings. It was demonstrated that short-term co-cultures, specifically performed with transient contact between the two cell types, were able to induce a transcriptional remodeling of Skov3 cells as shown by the up-regulation of ALDH1A3, IL1β, PDPN and MT1E transcripts. Moreover, under the same settings, Skov3 cells co-cultured with HG-SOC-MSCs but not N-MSCs nor when cultured in the presence of HG-SOC-MSCs conditioned medium, were reprogrammed as assessed by specific functional assays, highlighting an increased viability, not linked to proliferation, and higher tumorigenicity and motility. Whole genome expression analysis performed on Skov3 cells subjected to long-term co-culture experiments (5 days), allowed us to focus on different up-regulated transcripts, such as IL8, CCL2, CXCL1, CXCL2, TNF (involved in the interleukin-10 signaling pathway), CTGF and CYR61 (members of the CCN family), in addition to FOS, EGR1 and ATF4. Such transcripts are specifically up-regulated in Skov3 cells upon direct co-culture with HG-SOC-MSCs but not N-MSCs or in membrane-separated co-culture experiments. Furthermore, subcutaneous injection of an admix of Skov3 cells and HG-SOC-MSCs in NOD-SCID mice displayed an increased kinetics of the Skov3-derived tumor xenograft, showing that HG-SOC-MSCs are able to shorten the latency period of the tumor formation.

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RUSSO, SPENA CONCETTA. "Pin1 protein: a druggable target in high grade serous ovarian cancer." Doctoral thesis, Università degli Studi di Trieste, 2019. http://hdl.handle.net/11368/2957167.

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I focused my attention on Pin1 expression and chemical inhibition in HGS-EOC models. First, I confirmed that Pin1 is effectively involved in tumor progression utilizing mouse ovarian surface epithelial cancer cell line (STOSE), which closely recapitulates the characteristics of human HGS-EOC. So, I showed that comparing normal and knock down cells there is a tumor formation only in mice with Pin1 expression and activity. Then I focused my attention in order to develop an efficient Pin1 inhibitor, able to down regulate Pin1 expression in vitro but also in vivo. Starting from the inhibitor developed by Pfizer (Fig.9), which showed a good enzymatic activity in vitro 90, and it’s very specific for the catalytic site of the enzyme, in the last 3 years I worked in order to increase the permeability of this compound.The presence of carboxylate group permits the formation of different H-bond interactions in the proline binding site, interactions necessary for Pin1 catalytic function. Also the benzyl-imidaziole is a donator of hydrogen bonds that stabilize the interaction with Pin1. Despite significant improvements in Pin1 inhibitor affinity, these benzimidazole-based inhibitors failed to show cellular effects (up to 100 μM) on cancer cells. The authors hypothesized that this was due to poor cell permeability caused by high polarity on the benzimidazole series. The hydrophobic and nonionizable drugs cannot be loaded into liposomes through conventional means. In fact, ionisable hydrophilic drug can be remote loaded inside the liposomes using a transmembrane pH with efficient incorporation The most important example is the Doxil. But a poorly soluble hydrophobic drug is not encorporated into liposomes with the same high efficacy. For this reasons, how reported from Volgestein group, the hydrophobic compound can be actively loaded into liposomes by encapsulating them into modified cyclodextrins.The encapsulation of a poorly soluble drug into an ionizable cyclodextrin (preloading) enhances its water solubility and permits efficient liposomal loading via a pH gradient. In our protocol the Pin1 inhibitor, called compound 8, is preloaded inside modified cyclodextrins, Heptakis 6ammino6deoxy cyclodextrins, and then the complex cyclodextrin-compound 8 (C8) is loaded inside the liposomes via a pH gradient. How reported in the published article, the complex liposomes-cyclodextrin-compound 8 (LC8) is able to down regulate the Pin1 level in vitro, reducing the vitality of cancer cells, and also had activity in vivo, reducing the tumor volume and the Pin1 expression.

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Khalique, Lalarukh. "A Molecular Study of Clonality and Heterogeneity in High-Grade Serous Epithelial Ovarian Cancer." Thesis, Queen Mary, University of London, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.518266.

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TIERNO, DOMENICO. "Comprehensive Characterization and Effective Combinatorial Targeting of Epithelial Ovarian Cancer and High-Grade Serous Ovarian Cancer via Single-cell Analysis." Doctoral thesis, Università degli Studi di Trieste, 2021. http://hdl.handle.net/11368/2996074.

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Ovarian cancer kills more than 40 000 women in Europe and more than 150 000 women globally each year. The epithelial ovarian cancer (EOC) is the most common subtype and encompasses a collection of neoplasms with distinct clinical-pathological, molecular features and prognosis; among them the most common are the clear cells (CCOC), the mucinous (MOC), the endometrioid (ENOC), the low grade serous (LGSOC) and the high grade serous ovarian cancer (HGSOC). The HGSOC, in particular, is the most deadly form of EOC due to its high intratumor heterogeneity and lack of early diagnosis. The mechanical properties can be useful as new possible biomarkers of EOC subtypes in association to the classic ones used in clinical routine. In this thesis, the mechanical characterization of 9 ovarian cancer cell lines with different histological and morphological classification was carried out by AFM. Then, the achieved mechanical properties were evaluated on the same cell lines as markers of metastatic potential and drug sensitivity against 2C, a compound synthesized by the group of prof. Benedetti at the University of Trieste with anti-tumoral and cytoskeleton depolymerizing activity. Finally, an assessment of the effects of the culture medium composition on mechanical properties was performed. The mechanical characterization showed that the HGSOC cell lines had a high variability in the average Young’s modulus and resulted stiffer than the other cell lines with different histological classification. Moreover, the cell lines displayed two distinct single cell Young’s moduli distribution patterns: unimodal and bimodal. The cell lines with bimodal pattern showed two different populations with distinct mechanical behaviors. The invasion assay indicated a correlation between the stiffness decrease and the increase of invasion capacity. Accordingly, in cell lines with bimodal pattern only the “softer” population showed eventually the metastatic potential to invade. The cell lines with bimodal pattern were more resistant to 2C than the ones with unimodal pattern. For cell lines with bimodal pattern, the “stiffer” population had tendentially a higher resistance to 2C than the “softer” one. The F-actin network organization could influence the 2C resistance and the stiffness of cell lines: the HEY and OVCAR4 (high 2C resistance and high average Young’s modulus) had an Actin cytoskeleton more distributed over the cell than the TYKNU (low 2C resistance and low average Young’s modulus). Finally, variations in the culture medium components had an impact on the achieved Young’s moduli. This highlighted the need to develop optimized culture protocols for elasticity measurements, able to overcome the effects of different media on the mechanical properties.
Ovarian cancer kills more than 40 000 women in Europe and more than 150 000 women globally each year. The epithelial ovarian cancer (EOC) is the most common subtype and encompasses a collection of neoplasms with distinct clinical-pathological, molecular features and prognosis; among them the most common are the clear cells (CCOC), the mucinous (MOC), the endometrioid (ENOC), the low grade serous (LGSOC) and the high grade serous ovarian cancer (HGSOC). The HGSOC, in particular, is the most deadly form of EOC due to its high intratumor heterogeneity and lack of early diagnosis. The mechanical properties can be useful as new possible biomarkers of EOC subtypes in association to the classic ones used in clinical routine. In this thesis, the mechanical characterization of 9 ovarian cancer cell lines with different histological and morphological classification was carried out by AFM. Then, the achieved mechanical properties were evaluated on the same cell lines as markers of metastatic potential and drug sensitivity against 2C, a compound synthesized by the group of prof. Benedetti at the University of Trieste with anti-tumoral and cytoskeleton depolymerizing activity. Finally, an assessment of the effects of the culture medium composition on mechanical properties was performed. The mechanical characterization showed that the HGSOC cell lines had a high variability in the average Young’s modulus and resulted stiffer than the other cell lines with different histological classification. Moreover, the cell lines displayed two distinct single cell Young’s moduli distribution patterns: unimodal and bimodal. The cell lines with bimodal pattern showed two different populations with distinct mechanical behaviors. The invasion assay indicated a correlation between the stiffness decrease and the increase of invasion capacity. Accordingly, in cell lines with bimodal pattern only the “softer” population showed eventually the metastatic potential to invade. The cell lines with bimodal pattern were more resistant to 2C than the ones with unimodal pattern. For cell lines with bimodal pattern, the “stiffer” population had tendentially a higher resistance to 2C than the “softer” one. The F-actin network organization could influence the 2C resistance and the stiffness of cell lines: the HEY and OVCAR4 (high 2C resistance and high average Young’s modulus) had an Actin cytoskeleton more distributed over the cell than the TYKNU (low 2C resistance and low average Young’s modulus). Finally, variations in the culture medium components had an impact on the achieved Young’s moduli. This highlighted the need to develop optimized culture protocols for elasticity measurements, able to overcome the effects of different media on the mechanical properties.

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Wang, Yiying, Lingmin Li, Yue Wang, Zeng Yuan, Wenjing Zhang, Kenneth Hatch, and Wenxin Zheng. "IMP3 as a cytoplasmic biomarker for early serous tubal carcinogenesis." BioMed Central, 2014. http://hdl.handle.net/10150/610179.

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BACKGROUND:Serous tubal intraepithelial carcinoma (STIC) and the p53 signature in tubal mucosa have been supported to be precursor lesions in high-grade serous carcinoma (HGSC) of the fallopian tube, ovary, and peritoneum. It remains critical to find biomarkers for precursor lesions in order to detect HGSCs efficiently. IMP3 is an oncoprotein that has been explored in human malignancies. No studies have specifically addressed the expression of IMP3 in precursor or early lesions of HGSC. The main purposes of this study are to evaluate if IMP3 plays any role in the process of pelvic serous carcinogenesis by examining its expression in HGSC precursor lesions, to examine the relationship between IMP3 and p53 in those precursor lesions, and to check if IMP3 can be used as a biomarker for early diagnosis.METHODS:Immunohistochemistry for IMP3 and p53 was performed and evaluated in 48 HGSCs with STIC, 62 HGSCs without STIC, and 60 benign cases as negative controls. Sections of fallopian tubes with or without STIC , as well as cancers within the ovaries, were studied. IMP3 signature was defined as strong IMP3 cytoplasmic staining in 10 or more consecutive benign-looking tubal epithelial cells. The relationship between IMP3 and p53 overexpression was examined.RESULTS:In the 48 HGSC patients with STIC, IMP3 was positive in 46% of STIC lesions and had a similar positive rate in the invasive components of HGSC. IMP3 was also expressed in normal appearing tubal epithelia (IMP3 signature) in 15 (31%) of 48 HGSC cases with STIC and 10 (16%) of 62 cases without STIC. In contrast, no single IMP3 signature was found in the benign control group. Concordant expression of IMP3 and p53 signatures in the STIC group was found in up to one-third of the cases. There were also five (10%) STIC cases with positive IMP3 and negative p53.CONCLUSIONS:We conclude that IMP3 may be involved in the process and progression of pelvic HGSC and may serve as a complimentary biomarker in diagnosing STIC.

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Farquharson, Malcolm John. "Improving the understanding of platinum sensitivity and the tumour microenvironment in high grade serous ovarian cancer." Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/31006/.

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Ovarian cancer is one of the most lethal malignancies and often presents at an advanced stage, resulting in a poor prognostic outlook. Platinum chemotherapy leads to an initial clinical response, however most patients will ultimately relapse and there remains a sub-group who are intrinsically resistant to platinum. I focussed on high grade serous ovarian cancer (HGSOC), the most common subtype of ovarian cancer. The ID8 CRISPR-generated models represented a novel and simple tool to investigate the biology of HGSOC. By using this in vivo model, I aimed to further the understanding of platinum sensitivity in HGSOC by investigating the homologous recombination pathway and the tumour microenvironment. In vitro work showed that sensitivity to PARP inhibitors was clearly correlated with defective homologous recombination but the relationship with platinum sensitivity was more complicated. Using the ID8 derivatives, in vivo cisplatin experiments identified Pten and Nf1 loss to be associated with the worst prognosis with the knockout of Brca1 or Brca2 prolonging survival. A Brca1 mutation in the PALB2 domain compared to the BRCT2 domain was found to be associated with a greater sensitivity to cisplatin. The tumour microenvironment was shown to differ between genotypes and altered with the addition of platinum chemotherapy. Specifically, the loss of Pten was associated with an immunosuppressive microenvironment with increased levels of myeloid-derived suppressor cells (MDSCs) and tumour-associated macrophages (TAMs). The chemokines, Ccl2 and Ccl7 were shown to be significantly increased in the Trp53-/-;Pten-/- genotype. I targeted both the cytokine/chemokine response directly by using a transgenic mouse model (CCR1, 2, 3, 5 receptors knockout) and the PI3K/AKT pathway by using a PI3K inhibitor (p110β) (AZD8186) to attempt to reverse the effect of Pten loss. The transgenic mouse model (GGTACKO) showed encouraging early results with a reduction in MDSCs and TAMs in the knockout mice injected with the Trp53-/-;Pten-/- genotype but a repeat experiment is required before valid conclusions can be made. The AZD8186 in vivo experiment showed a significant reduction in MDSC levels in the ascites following AZD8186 treatment in mice injected with the Trp53-/; Pten-/- genotype and a non-significant decrease in the tumour samples. There was also a reversal in the anaemia previously shown with the loss of Pten and a decrease in Ccl2 and Ccl7 expression. I have used a transplantable in vivo model for HGSOC to investigate potential mechanisms of platinum sensitivity and identified poor prognostic genotypes (Pten, Nf1). I have found Pten loss to be associated with an immunosuppressive microenvironment and highlighted potential therapeutic targets. By targeting the PI3K/AKT pathway I have shown that the effect of Pten loss can be reversed. The next step will be to determine whether this reversal results in a prolonged survival.

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AZZALINI, EROS. "Comprehensive characterization and effective combinatorial targeting of high-grade serous ovarian cancer via single-cell analysis." Doctoral thesis, Università degli Studi di Trieste, 2020. http://hdl.handle.net/11368/2967979.

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Ovarian cancer kills more than 40 000 women in Europe and more than 150 000 women globally each year. High-grade serous ovarian cancer (HGSOC) is the most common and most difficult to treat subtype of the disease. The high-grade serous tumors are highly heterogeneous, therefore, though most of the patients respond well to surgery and chemotherapy initially, more than half experience relapse. HERCULES is a research project funded by the EU H2020 program with the target to characterize comprehensively high grade serous ovarian cancers to find novel therapeutic strategies to fight them. The aim of my PhD thesis was to validate biomarkers identified within HERCULES project in a retrospective case study of patients affected by HGSOC, studying tumor heterogeneity and evaluating the effects of pre-analytical variables, in particular fixation, in the validation process. High grade serous ovarian cancer samples were characterized validating selected biomarkers at both RNA and protein level. Molecular analysis and in situ analysis were performed on multiple tissue biopsies in order to detect spatial heterogeneity and, moreover, biomechanical proprieties of fixed tumor tissues were measured. Lastly, the reliability of molecular analyses on archive tissues were assessed, determining the effect of formalin and Bouin’s fixation at RNA level and evaluating their impact on gene expression using different platforms. Our results showed that detail morphological and immunophenotypical analyses, at the level of the entire tissue slide and not of TMA spots (Tissue micro array), of HGSOC tumors are paramount for the differential diagnosis as well as for both prognostication and therapy. In this view, biomechanical properties, by AFM can support the morphological findings. Among the immunohistochemical markers, Ki67 and BRCA1 have been shown their predictive value for response to first line chemotherapy and overall survival in HGSOC patients. Furthermore, neo adjuvant chemotherapy seems to have a detrimental effect on patient in our cohort. At the RNA level, cyclin C and HLA-B biomarkers showed their prognostic value indicating longer overall survival, while AKTs isoforms have shown a different impact on patients’ outcome. Regarding the pre-analytical variables, fixation confirmed to have a deep impact on molecular analyses, especially in RNA expression. Tissues with highly fragmented RNA such those fixed in Bouin’s can lead to analytical bias in both ddPCR, RT-qPCR, Nanostring and RNAscope technologies. A careful selection of samples with proper nucleic acids quality and integrity is of paramount importance before starting any molecular analysis. Also in that case, to minimize the effect of sample to sample variability a proper sample size should be used. Bouin’s fixed samples because of their high level of nucleic acids fragmentation are not recommended for mRNA expression analyses, especially for low expressed targets. Contrarily, miRNAs, giving their length, are more resistant to fixation procedures and can be used for RNA expression analyses in both formalin and Bouin’s tissues after a proper method of normalization

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Chen, Hao, Robert Klein, Stacy Arnold, Setsuko Chambers, and Wenxin Zheng. "Cytologic studies of the fallopian tube in patients undergoing salpingo-oophorectomy." BIOMED CENTRAL LTD, 2016. http://hdl.handle.net/10150/621541.

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Background: Mounting evidence suggests the fallopian tube as the origin for ovarian high grade serous carcinoma (HGSC). We attempted to identify the tubal cytological features that allow us to distinguish malignant from benign conditions. Methods: Tubal specimens (n = 56) were collected from patients who underwent bilateral salpingo-oophorectomy (BSO) due to various clinical indications. A standard procedure to collect fallopian tube brushings from freshly received surgical specimens was developed. Cytological diagnoses were classified into three categories: benign, atypical, and suspicious for malignancy/malignant. Cytological variables of individual cells and epithelia were subjected to statistical analysis. The fallopian tube histology was used as diagnostic reference for confirmation of cytology diagnosis. Results: Among the 56 fallopian tube specimens, 2 (3.7 %) showed inadequate cellularity preventing further evaluation, 11 (20.4 %) were diagnosed as malignant or suspicious of malignancy, 7 were atypical, and 36 were benign. The presence of three dimensional clusters (p < 0.0001, Fisher's Exact Test), or prominent nucleoli (p = 0.0252, Fisher Exact test) was highly correlated with the diagnosis of malignancy. The suspicious malignant/malignant cytological diagnosis was also highly correlated with presence of HGSC with or without serous tubal intraepithelial carcinoma (STIC). Conclusions: Tubal cytology may be useful for ovarian cancer screening and early detection.

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LUONGO, RAFFAELE. "TOWARDS PATIENT-SPECIFIC MODELS IN HIGH GRADE SEROUS OVARIAN CANCER (HGSOC): LINKING EPIGENETIC TRACING OF CELL OF ORIGIN WITH ACTIONABLE ORGANOID MODELS." Doctoral thesis, Università degli Studi di Milano, 2021. http://hdl.handle.net/2434/883835.

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Ovarian Cancer (OC) is a major cause of cancer-related mortality, due to the late-stage diagnosis and failure of surgery and chemotherapy to fully eradicate the disease, that is reflected in a high rate of tumor relapse after treatments. Patients with high-grade serous ovarian cancer (HGSOC), representing the largest majority of OC (~70%), have not experienced significant improvement in overall survival in last decades (1), pointing to the acute need to identify new predictive biomarkers and therapeutic targets for clinical settings. This unresolved emergency derives from our poor understanding of HGSOC biology combined with the recognized lack of suitable clinically relevant models for this disease, as the available models fail to relate molecular aberrations to clinical histories (2). Moreover, one major challenge that sets HGSOC apart from all other solid tumors, that has witnessed significant progress in recent years, is the persistent uncertainty about its cell of origin and the consequent lack of molecular signatures for the unequivocal assignment of specific samples to either an ovarian surface epithelium (OSE) or a fallopian tube fimbrial epithelium (FI) origin, the two candidate tissues still debated in literature (3, 4). Altogether, these aspects have hampered the dissection of pathogenic mechanisms and the identification of targets for improved clinical care of patients, driving us towards the embracement of new paradigms for effective advancements in the clinical setting. To overcome these issues, in the past few years, in the lab, we have devoted our efforts to the identification of the cell of origin of the tumor and the development of a clinical model allowing the study of this disease. Our applications aimed at brought a valid boost towards those precision oncology approaches that promises to transform cancer care by delivering personalized treatments tailored to the genetic and epigenetic specificities of patients’ tumors, whose remarkable heterogeneity characterizes even seemingly homogeneous histotypes. This level of complexity urged the need to develop and validate patient-derived cancer models that recapitulate to a meaningful extent the features of primary or metastatic tumors, both for efficient drug development and for guiding therapeutic strategies. Among new technologies, patient-derived organoid cultures are emerging as powerful models able to recapitulate in a meaningful extent the features of primary tumors (5, 6). Their value was proved both in guiding therapeutic decision for broadly curable tumors (7) and in gaining new insights for intractable tumors such as pancreatic adenocarcinoma (8). OC studies have started to benefit from these advanced models only recently (9–11), and a huge part of this PhD work regards the generation of organoids models from both HGSOC and normal samples of FI and OSE, in order to encompass all kind of tissues useful for the study of the pathology. Indeed, while tumor organoids represent an in vitro proxy of OC at an advanced stage, normal organoids from healthy tissues can be used as control but also as an indefinitely expandable cellular model in which investigate tumor alterations in the context of a normal tissue. This is really significant also in light of our recent publications on the cell of origin of HGSOC (12), where we showed the establishment of a method that is able to stratify HGSOC tumors through an approach based on methylomics and transcriptomics that: i) allows to distinguish OSE- from FI- derived tumors; ii) allows to identify for each tumor subtype the specific transcriptomic alterations that could drive the tumorigenic process; iii) reveals a prognostic value for the tissue of origin of this disease. Taken together, these two sets of information could help in define the molecular mechanisms governing the disease at an unprecedented resolution, setting a paradigm for harnessing patient-specific cancer models in the complementary goal of dissecting pathogenic mechanisms and extracting their actionable features. The extrapolation of information from organoids taking in consideration the cell of origin, through both the increase of our knowledge of the pathology and the identification of new markers and targets to be used as novel drug therapies, will prospectively pave the way to the improve the clinical care of patients with HGSOC.

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Yamanoi, Kouji. "Suppression of ABHD2, identified through a functional genomics screen, causes anoikis resistance, chemoresistance and poor prognosis in ovarian cancer." Kyoto University, 2017. http://hdl.handle.net/2433/227585.

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Silva, Munasinghage Lakmali. "Determination of the kallikrein-related peptidase 7 degradome and Transciptome in ovarian cancer." Thesis, Queensland University of Technology, 2016. https://eprints.qut.edu.au/92869/1/Munasinghage%20Lakmali_Silva_Thesis.pdf.

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Ovarian cancer is the leading cause of cancer-related death in women, and the need for curative treatments is urgent. This study characterised an enzyme associated with the most lethal form of ovarian cancer, showing this enzyme to be a promising therapeutic target. Fifteen novel protein targets and key signalling pathways were determined to be regulated by this enzyme, kallikrein-related peptidase 7, in the ovarian tumour microenvironment, highlighting its involvement in cancer progression. Inhibition of this enzyme may be a useful therapeutic option to improve the life expectancy of women suffering from this cancer.

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Zickgraf, Franziska Maria [Verfasser], and Andreas [Akademischer Betreuer] Trumpp. "SPDEF is a mediator of tumorigenicity in SSEA1- tumor-initiating cells in high grade serous ovarian cancer / Franziska Maria Zickgraf ; Betreuer: Andreas Trumpp." Heidelberg : Universitätsbibliothek Heidelberg, 2021. http://d-nb.info/1230067167/34.

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Jiménez, Sánchez Alejandro. "Characterisation of the tumour microenvironment in ovarian cancer." Thesis, University of Cambridge, 2019. https://www.repository.cam.ac.uk/handle/1810/287935.

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The tumour microenvironment comprises the non-cancerous cells present in the tumour mass (fibroblasts, endothelial, and immune cells), as well as signalling molecules and extracellular matrix. Tumour growth, invasion, metastasis, and response to therapy are influenced by the tumour microenvironment. Therefore, characterising the cellular and molecular components of the tumour microenvironment, and understanding how they influence tumour progression, represent a crucial aim for the success of cancer therapies. High-grade serous ovarian cancer provides an excellent opportunity to systematically study the tumour microenvironment due to its clinical presentation of advanced disseminated disease and debulking surgery being standard of care. This thesis first presents a case report of a long-term survivor (>10 years) of metastatic high-grade serous ovarian cancer who exhibited concomitant regression/progression of the metastatic lesions (5 samples). We found that progressing metastases were characterized by immune cell exclusion, whereas regressing metastases were infiltrated by CD8+ and CD4+ T cells. Through a T cell - neoepitope challenge assay we demonstrated that pre- dicted neoepitopes were recognised by the CD8+ T cells obtained from blood drawn from the patient, suggesting that regressing tumours were subjected to immune attack. Immune excluded tumours presented a higher expression of immunosuppressive Wnt signalling, while infiltrated tumours showed a higher expression of the T cell chemoattractant CXCL9 and evidence of immunoediting. These findings suggest that multiple distinct tumour immune microenvironments can co-exist within a single individual and may explain in part the hetero- geneous fates of metastatic lesions often observed in the clinic post-therapy. Second, this thesis explores the prevalence of intra-patient tumour microenvironment het- erogeneity in high-grade serous ovarian cancer at diagnosis (38 samples from 8 patients), as well as the effect of chemotherapy on the tumour microenvironment (80 paired samples from 40 patients). Whole transcriptome analysis and image-based quantification of T cells from treatment-naive tumours revealed highly prevalent variability in immune signalling and distinct immune microenvironments co-existing within the same individuals at diagnosis. ConsensusTME, a method that generates consensus immune and stromal cell gene signatures by intersecting state-of-the-art deconvolution methods that predict immune cell populations using bulk RNA data was developed. ConsensusTME improved accuracy and sensitivity of T cell and leukocyte deconvolutions in ovarian cancer samples. As previously observed in the case report, Wnt signalling expression positively correlated with immune cell exclusion. To evaluate the effect of chemotherapy on the tumour microenvironment, we compared site-matched and site-unmatched tumours before and after neoadjuvant chemotherapy. Site- matched samples showed increased cytotoxic immune activation and oligoclonal expansion of T cells after chemotherapy, unlike site-unmatched samples where heterogeneity could not be accounted for. In addition, low levels of immune activation pre-chemotherapy were found to be correlated with immune activation upon chemotherapy treatment. These results cor- roborate that the tumour-immune interface in advanced high-grade serous ovarian cancer is intrinsically heterogeneous, and that chemotherapy induces an immunogenic effect mediated by cytotoxic cells. Finally, the different deconvolution methods were benchmarked along with ConsensusTME in a pan-cancer setting by comparing deconvolution scores to DNA-based purity scores, leukocyte methylation data, and tumour infiltrating lymphocyte counts from image analysis. In so far as it has been benchmarked, unlike the other methods, ConsensusTME performs consistently among the top three methods across cancer-related benchmarks. Additionally, ConsensusTME provides a dynamic and evolvable framework that can integrate newer de- convolution tools and benchmark their performance against itself, thus generating an ever updated version. Overall, this thesis presents a systematic characterisation of the tumour microenvironment of high grade serous ovarian cancer in treatment-naive and chemotherapy treated samples, and puts forward the development of an integrative computational method for the systematic analysis of the tumour microenvironment of different tumour types using bulk RNA data.

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Garnier, Camille. "Rôle de la protéine MAP3K8 et impact de la rigidité dans les cancers ovariens sereux de haut grade." Thesis, Sorbonne Paris Cité, 2016. http://www.theses.fr/2016USPCC223/document.

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Les cancers ovariens, se développant de façon silencieuse, diagnostiqués à des stades tardifs et de mauvais pronostiques, requièrent urgemment la mise au point de nouvelles options thérapeutiques. Ma thèse s'est attachée à caractériser les propriétés physiques et biologiques des cancers ovariens Séreux de Haut Grade (HGSOC), représentant 75% des tumeurs ovariennes.En premier lieu, nous avons démontré la valeur pronostique de la protéine MAP3K8 s'accumulant dans les HGSOC. Nous avons montré que MAP3K8 contrôle la prolifération et la migration des cellules cancéreuses via la transition G 1/S et les mécanismes d'adhésion dynamique. Aussi, nous avons mis en évidence que MAP3K8 active majoritairement la voie MEK, présentant ainsi un potentiel prédictif des inhibiteurs de MEK, les positionnant comme une stratégie thérapeutique prometteuse, en combinaison des thérapies conventionnelles, chez les HGSOC.Dans un second axe de ma thèse, nous avons montré que la rigidité augmente avec la taille tumorale, chez les HGSOC présentant une signature moléculaire « Fibrose ». Cette rigidification tumorale s'associe à une accumulation de stroma et un remodelage du réseau de collagène, mais aussi à une activation spécifique de la voie MEK. De façon intéressante, la rigidification tumorale accompagne un « switch » métabolique glycolytique, restreint au centre de la tumeur, la périphérie demeurant plus molle, et différant par une production élevée de collagène et un métabolisme OXPHOS. La rigidité pourrait donc être au carrefour de 3 processus majeurs, tels un remodelage de la matrice, l'activation de MEK et un switch métabolique stromal, expliquant, au moins en partie, la progression des HGSOC
Ovarian cancers, which develop in a silent manner in the peritoneal cavity, resulting in a late diagnosis and a poor prognosis, urgently require new therapeutic strategies. In this context, my thesis aimed at better characterize the physical and biological properties of the High Grade Serous ovarian cancers (HGSOCs), accounting for 75% of the tumours.First, we found that the protein MAP3K8 accumulates in HGSOC and is a potential prognostic marker for these tumours. We demonstrated that MAP3K8 controls cancer cell proliferation and migration by regulating key players in Gl/S transition and adhesion dynamics. Importantly, we highlighted that MAP3K8 function is mainly mediated by the MEK pathway, and exhibits a predictive potential for MEK inhibitors, defining them as a promising therapeutic option, in combination with conventional therapy, for HGSOC patients.In a second part of my thesis, we showed that tumor stiffness is increased during tumor growth in HGSOC presenting a "Fibrosis" molecular signature. Moreover, tumor stiffening is associated with high stromal content and remodeling of the collagen network. Interestingly, the MEK kinase was specifically activated upon tumor stiffening. Furthermore, tumor stiffness accompanies a glycolytic metabolic switch, restricted to the central part of stiff tumors. Indeed, the periphery of stiff tumors remains softer than the central part with stromal cells secreting high levels of collagens and showing an OXPHOS metabolism. Thus, tumor stiffness could be at the crossroad of three major processes, i.e. matrix remodeling, MEK activation and stromal metabolic switch, that might explain, at least in part, the progression of HGSOC

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Liu, Yueyang [Verfasser], Viktor [Akademischer Betreuer] Magdolen, Birgit [Gutachter] Luber, and Viktor [Gutachter] Magdolen. "Kallikrein-related peptidase 10: a novel independent prognostic marker in advanced high-grade serous ovarian cancer and triple-negative breast cancer / Yueyang Liu ; Gutachter: Birgit Luber, Viktor Magdolen ; Betreuer: Viktor Magdolen." München : Universitätsbibliothek der TU München, 2019. http://d-nb.info/1183910061/34.

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Geng, Xiaocong [Verfasser], Viktor [Akademischer Betreuer] Magdolen, Birgit [Gutachter] Luber, and Viktor [Gutachter] Magdolen. "Clinical relevance of kallikrein-related peptidases in advanced high-grade serous ovarian (KLK11, KLK15) and in triple-negative breast cancer (KLK11) / Xiaocong Geng ; Gutachter: Birgit Luber, Viktor Magdolen ; Betreuer: Viktor Magdolen." München : Universitätsbibliothek der TU München, 2019. http://d-nb.info/1200098668/34.

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Gong, Weiwei [Verfasser], Viktor [Akademischer Betreuer] Magdolen, Karl-Friedrich [Gutachter] Becker, and Viktor [Gutachter] Magdolen. "Assessment of kallikrein-related peptidases 4, 5, 7 and 12 as prognostic biomarkers in advanced high-grade serous ovarian cancer and triple-negative breast cancer / Weiwei Gong ; Gutachter: Karl-Friedrich Becker, Viktor Magdolen ; Betreuer: Viktor Magdolen." München : Universitätsbibliothek der TU München, 2020. http://d-nb.info/1213898986/34.

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Chen, Shuhui. "Étude des mutations des gènes KRAS, NRAS, BRAF, PIK3CA, MET et de l’expression des protéines P53 et PTEN et leurs implications cliniques dans le carcinome ovarien de haut grade." Thesis, Université de Lorraine, 2016. http://www.theses.fr/2016LORR0093/document.

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Objectifs: Malgré leur grande hétérogénéité histologique et moléculaire, la prise en charge clinique des carcinomes ovariens de haut-grade (COHG) reste peu variable. Le pronostic sombre de cette pathologie implique un réel besoin des nouvelles thérapies. Au-delà des marqueurs pronostiques histologiques classiques et des enquêtes oncogénétiques, l’objectif de cette étude a consisté à rechercher des cibles moléculaires pharmacologiquement recrutables afin de pouvoir proposer aux patientes un accès à la thérapie innovante et personnalisée. Méthodes: Cette étude a été réalisée chez 53 patientes (pts) (âge moyen 58,9 ans, intervalle 25-87) de COHG histologiquement prouvés dont 45 pts de sous-type séreux. 19 pts ont fait l’objet d’une consultation et d’un test oncogénétique sur la base d’antécédents familiaux / personnel de cancer de sein/ovaire. chez. L’expression de P53 et de PTEN a été évaluée sur des tissus fixés au formol et inclus en paraffine par immunohistochimie. Les mutations somatiques de KRAS, NRAS, BRAF, PIK3CA et MET ont été recherchées par PCR-HRM (Polymerase Chain Reaction High Resolution Melting) puis vérifiées par NGS (Next Generation Sequencing) sur des extraits d'ADN préparés à partir d'échantillons de tumeurs congelés, prélevés au moment du diagnostic. Résultats: Des mutations germinales de BRCA1 / 2 ont été identifiées chez 7 pts, toutes atteintes des carcinomes séreux. Une mutation du gène KRAS (exon 2), 2 mutations du gène NRAS (exon 3), 6 mutations du gène PIK3CA (exon 5, 10 et 21) et 5 mutations du gène MET (exon 14 et 18) ont été identifiées chez les 53 tumeurs par NGS, dont deux mutations du gène NRAS et 2 mutations du gène PIK3CA détectées précédemment par PCR-HRM. Aucun profil mutationnel multiple n’a été retrouvé. La surexpression de P53 et la perte d’expression de PTEN ont été constatées chez 32 sur 53 (60%) et 19 sur 46 (41%) des tumeurs. L’analyse statistique n’a été réalisée que chez le sous-groupe de pts atteintes des carcinomes séreux à cause de l’effectif de l’étude. Avec un suivi médian de 38 mois (intervalle de 6-93), 35 pts ont eu une rechute de la maladie et 25 pts sont décédées. La survie sans progression à 2 ans est 28%, et la survie globale à 5 ans est 37%. La surexpression de P53 a été trouvée associée à une meilleure chimiosensibilité, une meilleure survie sans progression et une meilleure survie globale. Conclusion: Pour des COHG, au-delà des altérations de P53 et PTEN, des anomalies génétiques somatiques concernant les voies de signalisation PI3K et MAPK ne sont pas rares et peuvent être détectées par NGS. L’identification de ces anomalies somatiques pourrait offrir une possibilité des thérapies ciblées innovantes pour les patientes sur la base d’éléments diagnostics moléculaires
Objectives: Despite the great histological and molecular heterogeneity, the clinical management of high-grade ovarian carcinoma remains univo-cal. As a major subgroup of ovarian carcinoma, high-grade ovarian carci-nomas (HGOC) need novel therapy. Additionally to conventional histolog-ical prognostic markers and oncogenetic investigations, molecular diag-nostic was performed using PCR-HRM (Polymerase Chain Reaction High Resolution Melting) and NGS (Next Generation Sequencing) to identify "druggable" targets that could provide access to innovative personalized therapy. Methods: This study was performed in 53 patients (pts) (mean age 58.9 years, range 25-87) with histologically proven HGOC of which 45 pts with serous carcinoma. BRCA1/2 germline mutations had been screened in 19 pts with familial/personal history of breast/ovarian cancer justifying on-cogenetic investigations. P53 and PTEN expression was assessed on for-malin fixed paraffin-embedded tissues using immunohistochemistry. So-matic mutations of KRAS, NRAS, BRAF, PIK3CA and MET were screened using PCR-HRM and then confirmed using NGS on DNA extracts from frozen tumor specimens taken at diagnosis. Results: Seven pts had BRCA1 / 2 germline mutations, all had serous carcinomas. One mutation of KRAS (exon 2), 2 mutations of NRAS (exon 3), 6 mutations of PIK3CA (exon 5, 10 and 21) and 5 mutations of MET (exon 14 and 18) were identified using NGS, of which 2 mutations of NRAS and 2 mutations de PIK3CA detected previously by PCR-HRM, no multiple mutation was detected. P53 overexpression and PTEN loss of expression was detected respectively in 32 of 53 (60%) and 19 of 46 (41%) of all the tumors. Because of the efffective of the study, statistical analyses were restricted to pts with serous carcinoma. With a median follow-up of 38 months (range 6-93), 35 pts had disease progression and 25 pts died during the follow-up. The 2-year progression-free survival (PFS) rate was 28% and 5-year overall survival (OS) rate was 37%. Overexpression of mutant P53 was found to be associated with chemosensitivity and longer PFS and OS. Conclusion: In HGOC, beside P53 and PTEN alterations, somatic genetic abnormalities of PI3K and MAPK signaling pathways can be detected us-ing NGS and provide molecular rationale for targeted therapies, potential-ly offering new therapeutic opportunities to the patients

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Milea, Anca. "Abrogation of the Retinoblastoma Pathway Defines Clinical Outcome in High Grade Serous Carcinoma." Thesis, 2012. http://hdl.handle.net/1807/42403.

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High Grade Serous Carcinoma is the most aggressive tubal/ovarian histotype. The Rb pathway, which functions to preserve cell cycle regulation and genomic stability, is frequently deregulated in HGSC. The aim of this study was to identify non-redundant mechanisms of Rb pathway deregulation with clinical relevance. Immunohistochemistry analysis of P16 and RB1 expression identified a P16 homogeneous/RB1+ subgroup with the shortest recurrence free survival, while P16 heterogeneous/RB1+ and P16 homogenous/RB1- subgroups showed longer recurrence free survival. These P16/RB1 categories were shared between precursor lesions and the associated HGSC. Characterization of the Rb pathway in the three subgroups revealed CCNE1 amplification and highest protein expression in the P16 homogeneous/RB1+, high cyclin D1 protein expression in P16 heterogeneous/RB1+, and E2F3 amplification and highest Ki67 protein expression in P16 homogeneous/RB1- subgroup. Rb pathway deregulation is common in HGSC, occurs early in tumor development, and results from multiple non-redundant mechanisms that correlate with clinical outcome.

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Chung, I.-Jiuan, and 鍾怡娟. "Determining Cooperative Lethal Effect of Targeted therapy in High-Grade Serous Ovarian Carcinoma." Thesis, 2019. http://ndltd.ncl.edu.tw/handle/txg6nv.

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Tone, Alicia. "The Role of Luteal Phase Fallopian Tube Epithelium in High-grade Ovarian Serous Carcinoma." Thesis, 2010. http://hdl.handle.net/1807/32954.

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Studies of prophylactic salpingectomy specimens from BRCA1/2 mutation carriers, at risk for tubal and ovarian high-grade serous carcinoma (SerCa), have consistently revealed occult carcinomas and putative histological cancer precursors in the distal fallopian tube epithelium (FTE), supporting the FTE as the source of SerCa. In this thesis I molecularly characterized and compared non-malignant FTE from mutation carriers (FTEb) and control patients (FTEn) to identify alterations that may predispose to malignant transformation. Gene expression profiling of laser capture microdissected FTEn, FTEb and SerCa indicated that SerCa have similar molecular profiles whether of presumed ovarian or tubal origin, supporting the notion they share a common cell of origin within the FTE. Furthermore, FTEb samples obtained during the post-ovulatory luteal phase showed gene expression profiles closely resembling SerCa samples, suggesting that the luteal phase milieu may contribute to serous carcinogenesis. An initial hypothesis was that FTEb may respond differently to luteal progesterone compared to FTEn, via differential expression of progesterone receptor (PR) isoforms. However, similar relative isoform expression in FTEn and FTEb samples suggested that a luteal phase-associated factor other than progesterone directs gene expression changes in FTEb. The possibility that FTEb respond differently to ovulation-associated inflammatory cytokines that are locally elevated during the luteal phase was next investigated. Importantly, FTEb specimens previously found to cluster with SerCa based on their global gene expression profiles showed evidence of increased nuclear factor-κB (NFκB)-dependent (pro-inflammatory) signalling and diminished glucocorticoid receptor (GR)-dependent (anti-inflammatory) signalling. Furthermore, I demonstrate that disabled homolog 2 (DAB2), an adaptor molecule decreased in SerCa and FTE luteal samples, enhances both GR-mediated transactivation and suppression of NFκB signalling, implicating DAB2 as a crucial determinant of inflammatory signalling and ovarian cancer risk. Altogether, this thesis identifies gene expression changes in FTE from BRCA mutation carriers during the post-ovulatory luteal phase that parallel those detected in SerCa. The data support a proposed novel testable model for predisposing events contributing to SerCa that centres on an altered ability to quickly resolve the pro-inflammatory environment created by the ovulatory event.

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Yeo, Hsin Yueh, and 姚欣樂. "Investigating Cooperative Lethal Effect of EGFR and MDM2 Inhibitors on High-Grade Serous Ovarian Carcinoma." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/45108402529231371681.

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Alrubaish, Sarah. "Role of zinc transporter LIV-1 protein in high-grade serous ovarian cancer." Thèse, 2018. http://hdl.handle.net/1866/21364.

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Lin, Han-Wei, and 林漢威. "High Alpha-Folate Receptor Correlates with Poorer Survival in Serous Ovarian Carcinoma." Thesis, 2010. http://ndltd.ncl.edu.tw/handle/26149725544105240508.

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碩士
臺灣大學
分子醫學研究所
98
Objectives: To investigate the relationship between alpha-folate receptor (α-FR) expression levels to the clinico-pathologic features and outcomes of patients with serous ovarian carcinoma. Methods: Semi-quantitative reverse-transcription polymerase chain reaction detected α-FR expression in 91 patients with serous ovarian carcinoma. Clinico-pathologic parameters and biomarkers, including FIGO stage, tumor grade, optimal surgery, lymph nodes metastasis, pre-operative serum CA-125, and α-FR expression levels in cancerous tissues to evaluate disease-free interval (DFI) and overall survival (OS), were analyzed. Results: There were 4, 6, 65, and 16 patients with stages I, II, III, and IV ovarian carcinoma, respectively. Forty (44%) underwent optimal debulking surgery. Patients with advanced stages (stage I: 0.451, stage II: 0.415, stage III: 0.652, stage IV: 0.768, p=0.004) or those with sub-optimal debulking surgery (optimal: 0.490, sub-optimal: 0.766, p=0.003) had significantly higher α-FR expression levels compared to those with early stages or optimal debulking surgery. In prognostic analysis, high α-FR expression level (HR: 2.70 (1.20-6.05), p=0.016) was an independent poor prognostic factor for DFI and had a negative impact on OS with marginal significance (HR: 3.51 (0.93-13.29), p=0.065) using multivariate analyses. Conclusions: Patients of serous ovarian carcinoma with high α-FR expression in cancerous tissue have shorter DFI and OS. α-FR may be a potential biomarker for predicting the outcome of serous ovarian carcinoma patients. Key words: alpha-folate receptor, CA-125, ovarian serous carcinoma, survival

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ANGELICO, GIUSEPPE. "Aquaporin-1 expression as predictive marker of chemoresistance in ovarian high-grade serous carcinoma: a comparative study between preoperative peritoneal biopsies and surgical samples." Doctoral thesis, 2020. http://hdl.handle.net/11570/3181787.

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Introduction. Aquaporin 1 (AQP1), as component of transmembrane-water-channel family proteins, is involved in different aspects of cancer biology and metastatic events. AQPs expression has been documented in different human tumors and, recently, also in ovarian carcinoma (OC). Anyway, only few studies have investigated the prognostic role of AQP1 in OC; however, the relationship between AQP1 expression and response to chemotherapy remains to be fully elucidated. Materials and Methods. By immunohistochemistry, the expression of AQP1 was evaluated in the preoperative peritoneal biopsies of 32 patients with peritoneal carcinosis, in which a histological diagnosis of high grade serous ovarian carcinoma was made; a cut-off of >1 % positive tumor cells was utilized to distinguish immunoreactive positive cases from negative ones. Furtherly, the potential association between immunohistochemical expression of AQP1, chemotherapeutic response and clinico-pathological parameters, was statistically investigated. Results. Taking into consideration the cut-off, 20 (62.5%) cases showed positive membranous staining (AQP1+), while 12 (37.5%) cases were considered negative (AQP1-). In the AQP+ group, Fisher exact test showed a significant association of AQP1 expression with poor chemotherapy response in omental tissues complete response score (CRS) 1-2 (p= 0.0039), while a CRS 3 was never observed in all positive cases. No significant relationship emerged between AQP1 expression and other clinico-pathological variables. Discussion and conclusions. The study findings suggest that AQP1 may be expressed in a sub-group of OC cases and it could be proposed as predictive marker. Therefore, at the time of the diagnostic peritoneal biopsy, patients affected from high grade serous carcinoma should be categorized in 2 different predictive groups, such as AQP+ and AQP-; OC AQP+ cases may represent a subset of poor responders patients, that could be considered more eligible for cytoreductive surgery rather than for neoadjuvant chemotherapy.

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Langthasa, Jimpi. "A study of extracellular matrix dynamics in epithelial cancer progression with a special focus on ovarian cancer spheroidogenesis." Thesis, 2022. https://etd.iisc.ac.in/handle/2005/5979.

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Epithelial ovarian cancer (EOC) is one of the most debilitating gynecological cancers in women worldwide due to its insidious symptoms. The predominant subtype of ovarian cancer: high-grade serous ovarian carcinoma (HGSOC), is responsible for 75% of all fatalities associated with EOC. 90% of EOC patients have already reached an advanced stage of metastasis when they are diagnosed with the disease. Metastasis is frequently associated with ascites: an abnormal accumulation of fluid in the peritoneal cavity due to the occurrence of spheroids, clusters of disseminated malignant EOC cells. Spheroids contribute significantly to the morbidity and mortality associated with EOC. Despite this, the mechanisms associated with the formation of EOC spheroids are ill-understood. Investigations indicate intricate connections between these signaling modules with elements of reciprocal and hierarchical connections that underlie spheroidal morphogenesis and may provide insights into the identification of targets for future therapeutic strategies for EOC.

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WANG, KAI-HUNG, and 王凱弘. "Epigenetic Study in Gynecological Cancers: Discovery of New DNA Methylation Biomarker in Detecting Cervical Cancer and Downregulation of Tumor Suppressor Gene CNN1 in the Development of Ovarian High-Grade Serous Carcinoma." Thesis, 2017. http://ndltd.ncl.edu.tw/handle/f2xv8b.

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博士
慈濟大學
醫學科學研究所
105
Gynecologic cancers contain five main types: cervical, ovarian, uterine, vaginal, and vulvar, whereas the first 2 types are high morbidity and mortality. Except genetic changes, epigenetic alteration is also a common phenomenon in cancer development, which is used to downregulate the expression of tumor suppressor genes. DNA methylation is one of the mechanisms of epigenetic alterations. There are several DNA methylation biomarkers that had been identified by isolating the DNA from Pap smear in cervical cancer patients. However, the biomarker for detecting early stage of cervical is lack. Therefore, this study surveyed the cohorts from Buddhist Tzu Chi General Hospital, and tried to discover new DNA methylation biomarkers for early detection of cervical cancers. The results demonstrated that one non-clustered PCDH gene, PCDH10, and two clustered PCDH genes, PCDHA4 and PCDHA13 were hypermethylated since cervical intraepithelial neoplasia 2 (CIN2) stage. The methylation test of these PCDH genes is more specific but less sensitive than high risk human papilloma virus (HPV) test. In this regard, combination of DNA methylation and HPV test could generate new triage strategy for early detecting cervical and precancerous lesion. On the other hand, more and more evidences proved that the cell origin of ovarian high-grade serous carcinoma (HGSC) is from fallopian tube epithelium (FTE). However, the detail mechanisms of this process and HGSC carcinogenesis are largely unknown. Thus, our group generated HGSC cell model by immortalization of FTE cells, named FE25, and further transformation to HGSC-like cells, named FE-RAS. By using this cell model, this study examined the function of the putative tumor suppressor gene, Calponin h1 (CNN1). The results showed that CNN1 were downregulated in FE-RAS and malignant ovarian cancer cells as well as HGSC tissues. Overexpressed CNN1 colocalized with Actin stress fiber structure, leading cells in a flatten morphology. Overexpression of CNN1 in FE-RAS cells significantly reduced cell motility, invasiveness, tumorigenesis and increased anoikis and adhesion. Conversely, when knocking-down CNN1 in FE25 cells, the malignant properties were increased. Microarray analysis showed overexpression of CNN1 may upregulate the genes contained kinase and phosphatase activities, Integrin binding, and Actin biding but downregulate others which has chemokine activity. Indeed, Integrin α2 and Integrin β1 were positively expressed with CNN1, explaining why the flatten morphology formed and adhesion increased. Besides, this study found that the microenvironment of ovarian stroma favored the growth of FE-RAS cells, which fits the theory of “HGSC originated from FTE,” and overexpression of CNN1 could significantly inhibits FE-RAS migrating toward to ovarian microenvironment. In summary, this study identified new methylation markers, PCDH10, PCDHA4, and PCDHA13, for detecting early cervical cancers and demonstrated the tumor suppressor, CNN1, which has to be turned down before the tubal originating HGSC can exfoliate and survival the anoikis. This tumor suppressor mechanism is likely through the modification of cell scaffolds and the associated signal transductions. Integrin α2 and Integrin β1 are the important mediators.

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Jadhav, Rohit. "ROLE OF OVARIAN CANCER-INITIATING CELLS IN HIGH-GRADE SEROUS OVARIAN CARCINOGENESIS." Thesis, 2012. http://hdl.handle.net/1805/2773.

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Indiana University-Purdue University Indianapolis (IUPUI)
A subpopulation of tumor cells known as ovarian cancer initiating cells (OCICs) have been shown to be the cells that propagate the tumor phenotype in ovarian cancer. Studies have showed that a very small population (100) of these cells is sufficient to induce a tumor phenotype; while a large quantity of tumor cells (5 X 105) are required to induce such a phenotype. In this study we studied the functional changes in genes expressed in the OCIC phenotype which were important for such efficient propagation of cancers. To enable this analysis, we generated mRNA expression and DNA methylation profiles of OCICs and compared them with those of tumor and normal ovarian surface epithelial cells. We identified four pathways which regulated most of the observed changes and were predicted to be important factors in distinguishing the OCICs from tumors and normal cells. The gene signatures for these pathways were analyzed by unsupervised clustering in order to determine the similarities of OCICs with respect to tumor and normal samples. We further believed that the OCICs can be used as indicators towards the genesis and progression of early events in the ovarian cancers. In light of this, we considered two hypotheses which are currently addressing the genesis of ovarian cancer. The first hypothesis proposed ovarian surface epithelial cells to be cells of origin of the ovarian cancer while the other proposed the fallopian tube cells to be contributing the cell of origin for these cancers. It is also believed that these two cells can be reciprocal cells of origin for the cancer phenotype. In order to test these hypotheses, we integrated the in-house dataset with a public domain fallopian tube gene expression data. The integration of the results obtained from these analyses provided better understanding of the early events in ovarian carcinogenesis.

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Stewart, Jocelyn Melissa. "Organizing Cellular Heterogeneity in High-grade Serous Cancer." Thesis, 2013. http://hdl.handle.net/1807/36007.

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High-grade serous ovarian cancer (HG-SOC) is the most lethal gynecologic malignancy. Although most respond to initial therapy, the vast majority of patients eventually recur and die of their disease. Understanding intra-tumor cellular heterogeneity and inter-patient variability is necessary to effectively cure HG-SOC. The work described in this thesis should help to speed the progress of ovarian cancer research in several ways. First, I generated a robust xenograft model that recapitulates the cellular heterogeneity of HG-SOC. In addition, I performed gene expression profiling on a subset of xenografts and showed that they recapitulate the inter-patient diversity of this disease. Second, I applied this model to pre-clinical testing of a folate-targeted imaging agent and showed that it can identify metastatic studding by PET/CT and fluorescence imaging. Using my xenograft model, I investigated the properties of tumor-initiating cells (TIC) and demonstrated that TIC in HG-SOC are rare. Furthermore, although CD133 marks most TIC, heterogeneity in the phenotype is observed within individual tumors and between different patients. Finally, I used a transformative technology, CyTOF, to develop a novel pipeline for prioritization of candidate TIC markers, as well as for characterization of cellular heterogeneity in primary HG-SOC samples.

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Chen, Hsiang-Ju Helen, and 陳香儒. "The Investigation of the Role of GATA3 in High-Grade Serous Ovarian Cancer Stem Cells." Thesis, 2018. http://ndltd.ncl.edu.tw/handle/5xagny.

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博士
國防醫學院
生命科學研究所
106
Ovarian cancer (OC) is the most lethal gynecological malignancy. The novel diagnostic and prognostic biomarkers are urgently needed to aid in the prevention and management of ovarian cancer. However, all attempts to date, have not improve the overall survival rate of OC. To circumvent this limit, there are two approaches in this project were investigated. Firstly, identify DNA methylation biomarker for improvement of diagnosis and secondly, to identify cancer stem- specific factor for better prognosis. Unfortunately, studies of DNA methylation biomarkers were not successfully completed, but for the cancer-stem related studies, we had a more completed discoveries. For cancer stem related study, we analyzed differential transcriptomes at early differentiation of OC stem cells and identified the transcription factor GATA3 as highly expressed in stem, compared to progenitor cells. GATA3 recruit the H3K27me3 demethylase, UTX, activate stemness markers, and promote stem-like phenotypes in OC. Targeting UTX by its inhibitor, GSKJ4, impeded GATA3-driven stemness, and enhanced apoptosis of GATA3-expressing cancer cells. Combinations of gemcitabine or paclitaxel with GSKJ4, resulted in a synergistic cytotoxic effect. Our findings provide evidence for a new role for GATA3 in OC stemness, and demonstrate that GATA3 may serve as a biomarker for precision epigenetic therapy in the future.

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Calvo, Gonzalez Llilians. "Dissection moléculaire de la sénescence cellulaire induite par le stress et la thérapie dans le cancer de l’ovaire et son impact sur la réponse des patientes." Thèse, 2015. http://hdl.handle.net/1866/13853.

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Le cancer de l’ovaire (COv) est le cancer gynécologique le plus létal chez la femme et les traitements existants, chirurgie et chimiothérapie, ont peu évolué au cours des dernières décennies. Nous proposons que la compréhension des différents destins cellulaires tels que la sénescence que peuvent choisir les cellules du cancer de l’ovaire en réponse à la chimiothérapie pourrait conduire à de nouvelles opportunités thérapeutiques. La sénescence cellulaire a été largement associée à l’activité de la protéine TP53, qui est mutée dans plus de 90% des cas de cancer de l’ovaire séreux de haut grade (COv-SHG), la forme la plus commune de la maladie. Dans nos travaux, à partir d’échantillons dérivés de patientes, nous montrons que les cultures primaires du cancer de l’ovaire séreux de haut grade exposées au stress ou à des drogues utilisées en chimiothérapie entrent en senescence grâce à l’activité d’un isoforme du gène CDKN2A (p16INK4A). Dans ces cellules, nous avons évalué les caractéristiques fondamentales de la sénescence cellulaire tels que les altérations morphologiques, l’activité béta galactosidase associée à la sénescence, les dommages à l’ADN, l’arrêt du cycle cellulaire et le phénotype sécrétoire associé à la sénescence. En utilisant des micromatrices tissulaires construites à partir d’échantillons humains de COv-SHG pré- et post-chimiothérapie, accompagnées de leurs données cliniques, nous avons quantifié des marqueurs de sénescence incluant une diminution de la prolifération cellulaire quelques semaines après chimiothérapie. De façon intéressante, l’expression de p16INK4A dans les échantillons de COv-SHG prétraitement corrèle avec une survie prolongée des patientes suite au traitement. Ceci suggère ainsi pour la première fois un impact biologique bénéfique pour la présence de cellules cancéreuses qui sont capable d’activer la sénescence, particulièrement pour le traitement du cancer de l’ovaire. Dans le but de complémenter les thérapies actuelles avec des approches de manipulation pharmacologique de la sénescence, nos résultats suggèrent qu’il serait important de déterminer l’impact positif ou négatif de la sénescence induite par la thérapie sur la progression de la maladie et la survie, pour chaque type de cancer de façon indépendante.
Human ovarian cancer (OvCa) is the deadliest gynecologic malignancy and existing surgical/chemotherapeutic treatment options have been relatively static for decades. We propose that understanding OvCa cell fate decisions taken in response to chemotherapy could guide new therapeutic opportunities. Damage-induced cellular senescence is often associated with TP53 activity, which is heavily mutated in high grade serous (HGS) OvCa (>90%), the most common form of this disease. Here, using patient derived tissues, we show that primary HGS-OvCa cultures predominantly trigger CDKN2A- associated (p16INK4A isoform) senescence following exposure to stress or chemotherapy. Key senescence hallmarks including altered morphology, senescence-associated-Betagalactosidase, DNA damage, cell cycle arrest and the senescence-associated secretory phenotype were evaluated and detected in damaged cells. Using tissue microarrays built from pre- and post-treatment human HGS-OvC tissue samples with accompanying clinical data, we quantified post-treatment hallmarks of senescence including reduced cell proliferation weeks after chemotherapy. Importantly, p16INK4A expression in pretreatment HGS-OvC samples correlated with increased patients survival, suggesting for the first time that senescence-competence in human cancer cells may have a beneficial impact on treatment outcomes for patients. In order to guide the potential improvement of existing human therapies via pharmacological senescence manipulation, our results suggests that it is important to determine for many types of human cancer whether treatment-induced senescence positively or negatively impacts disease progression and patient survival.

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Ondič, Ondrej. "Praktická aplikace imunohistochemických a molekulárně - genetických metod v diferenciální diagnostice lézí urogenitálního a gynekologického traktu." Doctoral thesis, 2018. http://www.nusl.cz/ntk/nusl-391332.

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This thesis focuses on gynecopathology. It consists of a collection of seven papers published in pathology journals with impact factor. Introduction section contains selection of examples showing scientific application of molecular genetic methods. Further on the aims of individual research projects are described. The first project comprises histomophologic study of skin endometriosis addressing "mullerian" differentiation. A case report of a rare tumor namely borderline papillary serous tumor of the fimbriated end of the fallopian tube follows with molecular genetic analysis of KRAS, BRAF and p53 gene mutation status. Prospective longitudinal study on high grade squamous dysplasia (HSIL) of the cervix in HPV vaccinated women, so called DAV (dysplasia after vaccination), aims to elucidate pathogenesis of this phenomenon. Two other studies focus on incidence of fumarate hydratase deficient leiomyomas of the uterus and hereditary leiomyomatosis and renal cell carcinoma syndrome (HLRCC). The aim of those studies is to improve our diagnostic capability and increase detection rate of the patients with HLRCC syndrome. Finally a new subtype of HSIL namely bizarre cell dysplasia is described in two separate studies. Conclusion remarks contemplate the role of molecular genetics in surgical pathology.

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Fleury, Hubert. "Implication des inhibiteurs de PARP dans le cancer de l’ovaire." Thèse, 2017. http://hdl.handle.net/1866/20221.

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Dissertations / Theses on the topic 'High-grade serous ovarian carcinoma'

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