This thesis focuses on describing and investigating the experiences of caregivers of individuals with ovarian cancer. Caregivers are an essential part of cancer care, and yet they are not formally recognized as such. The special focus on these caregivers stems from the recognition that ovarian cancer is unique from more commonly studied diseases, with a poor prognosis and frequent recurrences. This thesis sought to study this understudied population. The thesis begins with a scoping review of existing literature that specifically investigated this population. From this study, it was confirmed that few studies had focused on this population, however the mapped literature suggested that these caregivers experienced significant compromises to their quality of life. Some preliminary studies identified a theme that the caregiving experience was influenced by the relationships with healthcare providers. This theme informed the second study of the thesis, that was a cross-sectional, correlational study that sought to recruit partner-caregivers of patients with ovarian cancer, a sample mostly of male-caregivers. This study sought to explore multiple facets of the caregiving experience as part of cancer care using the Cancer Caregiving Tasks, Consequences and Needs Questionnaire, measuring caregiver distress using the Hospital Anxiety and Depression Scale, and collecting sociodemographic and proxy-reports of the patient’s medical information. A total of 82 partner-caregivers were recruited for the study, and our sample were mostly men, White, affluent and highly educated. Most of their partners were diagnosed with stage III or IV disease, and were treated with both surgery and chemotherapy. This study’s analysis found that caregiving workload, lacking information from healthcare providers, problems with the quality of information and communication with healthcare providers, lacking time for social relations due to caregiving, and needing more help from healthcare providers correlated with distress outcomes. The third investigation sought to further explore these relationships by measuring attachment insecurity, as assessed by a short, modified version of Experiences in Close Relationships Scale. Using the same sample data, hierarchical regression analyses were used to test whether general attachment avoidance or attachment anxiety moderated the relationship between the caregiving experiences and distress outcomes. These analyses revealed that attachment anxiety contributed to a portion of the variance in distress, however the experiences with the healthcare team explained a large portion of the variance of distress. Attachment anxiety was found to play a minor role moderating the relationship between needing more help from healthcare providers and anxiety, and attachment avoidance contributed a very small, moderating role between lack of time for social relations and distress. Together, these studies have demonstrated that caregivers of patients with ovarian cancer are understudied, however they experience significant levels of depression and anxiety. Their distress is highly affected by their reported experiences as part of the cancer care team, regardless of their predisposition to distress through attachment insecurity.

Each gene or protein has its own function which, when combined with others, allows the group to perform more complex behaviors, e.g. carry out a particular cellular task (functional module) or affect a particular disease phenotype (disease module). One of the major challenges in systems biology is to reveal the roles of genes or proteins in functional modules or disease modules. In the first part of the dissertation, I present a data-driven method, Correlation Set Analysis (CSA), for comprehensively detecting active regulators in disease populations by integrating co-expression analysis and specific types of literature-derived causal relationships. Instead of investigating the co-expression level between regulators and their targets, I focus on coherence of regulatees of a regulator, e.g. downstream targets of a transcription factor. Using simulated datasets I show that my method can reach high true positive rate and true negative rate (>80%) even the regulatory relationships is weak (only 20% of regulatees are co-expressed). Using three separate real biological datasets I was able to recover well-known and as- yet undescribed, active regulators for each disease population. In the second part of the dissertation, I develop and apply a new computational algorithm for detecting modules of functionally related genes that are likely to drive malignant transformation. The algorithm takes as input the identity and locations of a small number of known oncogenes (a seed set) on a human genome functional linkage network (FLN). It then searches for a boundary surrounding a gene set encompassing the seed, such that the magnitude of the difference in linkage weights between interior-interior gene pairs, and interior-exterior gene pairs is maximized. Starting with small seed sets for breast and ovarian cancer, I successfully identify known and novel drivers in both cancer types. In the third part of the dissertation, I propose a module based approach for expanding manually curated functional modules. I use the KEGG pathway database as an example and the results show that my approach can successfully suggest both validated pathway members (genes that are assigned to a particular pathway by other manually curated pathway databases) and novel candidate pathway genes.

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第17813号 / 医博第3811号 / 新制||医||999(附属図書館) / 30628 / 京都大学大学院医学研究科医学専攻 / (主査)教授 髙折 晃史, 教授 武藤 学, 教授 杉田 昌彦 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DGAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第18895号 / 医博第4006号 / 新制||医||1009(附属図書館) / 31846 / 京都大学大学院医学研究科医学専攻 / (主査)教授 松田 道行, 教授 武藤 学, 教授 戸口田 淳也 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20253号 / 医博第4212号 / 新制||医||1020(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 河本 宏, 教授 戸井 雅和, 教授 小川 誠司 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

Oxidative stress (OS) occurs when DNA repair mechanisms are overcome by the amount of single and double strand DNA breaks caused by an accumulation of reactive oxygen species (ROS). Genomic instability (GI) by microsatellite instability (MSI) accumulation is characterized by changes in DNA single tandem repeats (STR) as a direct result of ROS. Deregulation of DNA repair and tumor suppressor pathways have been described as causes of tumor progression and metastasis. Studies in mammals have focused on GI and the implications of increased mutation frequency due to accumulation of MSI leading to development of diseases, including infertility and cancer. Ovarian cancer is a deadly disease displaying the highest mortality rate among gynecological cancers. Hereditary ovarian cancer displays GI that can be established early in primordial germinal cells (PCGs) development and migration across the genital ridge, where PGCs are exposed to ROS damage. The hypothesis of this study was ROS-induced GI is marked by the accumulation of MSI on repetitive sequences of DNA that override DNA repair, tumor suppressor and redox homeostasis pathways. In this study, we induced ROS in human ovarian cell lines by hydrogen peroxide (H2O2) exposure, as well as evaluated mouse PGCs to determine whether MSI occurs in specific regions of human and mouse genomes. Our results show that MSI was present in specific markers after ROS-induced damage in human ovarian cells and in mouse Sod1 knockout PGCs during cell migration, both of which accumulate specific mutations caused by free radical damage. Ovarian tumor cells and mouse PGCs showed an increase of MSI in 12 human and 5 mouse repetitive markers that are located near important genes related to DNA repair, tumor suppression, cell proliferation, apoptosis and differentiation. This could be a signal that leads to tumor initiation, formation and progression in adult ovarian cells due to improper DNA repair and tumor suppression mechanisms or in disruption of PGC migration that determines germinal cell pool selection during early embryonic development due to absence of cell antioxidant mechanisms. Therefore, these specific unstable STRs are novel biomarkers that could be useful in early diagnostics, prognosis, and successful therapy of ovarian tumorigenesis.

No description available.

The objective of this thesis is to examine the relationship between CAIX (a biomarker for insufficient oxygen in tumor microenvironment) and CD8+ T cells (the immune cells for killing cancer cells) for ovarian cancer. We approach the problem from two perspectives. The first approach is to set up count models such as Poisson, negative binomial, and zero-inflated Poisson models to examine the cell counts between CAIX and CD8+ T cells in the tumor microenvironment. The second approach is to apply the cross-K function, which is a second-order property of the point pattern process. We find that the tissue microarray (TMA), which is a technique to assemble hundreds of tissue samples on one TMA block, has a fixed effect on the CD8+ T cell counts. There are two TMA blocks A2 and B1. The relationship between CAIX and CD8+ T cells highly depends on TMAs. On TMA B1 stroma, a negative relationship between CAIX and CD8+ T cell counts is observed in the negative binomial models. When taking the spatial domain into account and comparing the estimated cross-K function of CAIX and CD8+ T cells to the simulated envelopes generated by a homogeneous Poisson process, we find that CAIX and CD8+ T cells are regulated and repel each other on TMA B1. Tissue category also plays an influential role in analyzing the relationship. The estimated cross-K function of CAIX and CD8 + T cells is more dispersed on tumors than on stroma. / Graduate

Ovarian cancer results in more deaths per year than any other cancer of the female reproductive system. The low survival rate is partly due to the lack of early detection and the susceptibility to relapse. The AKT serine threonine kinase plays a pivotal role in hallmark cellular processes for the progression of ovarian cancer, including tumor cell growth and migration. Therapeutic targeting of pan-AKT has been problematic, in part due to feedback mechanisms and crosstalk with other pathways. The hypothesis for this study is that AKT 1, -2 and -3 isoforms may have different roles and regulate cell processes in uniquely varied ways. A transgenic mouse model that expresses the SV40 T-antigen viral oncogene and is known to have dramatically increased susceptibility to ovarian cancer was utilized, and it had genetic inactivation of either AKT1 or AKT2 through targeted deletion of the individual genes because these isoforms have been implicated in this cancer. Primary ovarian tumor cell cultures were established and found to exhibit different morphology, proliferation and migration that may indicate a different role for the AKT1 and AKT2 isoforms in these contexts. Ovarian tumor cells with absence of AKT1 predominantly exhibited reduced cell migration when compared to cells with retention of AKT1 and absence of AKT2. Since AKT is known to be important for epithelial-mesenchymal transition (EMT), a process potentially associated with tumor cell metastasis, the expression of transcription factors implicated in EMT was assessed by real-time array analysis in ovarian tumor cells knocked-out for either AKT1 or AKT2. Twist1, one of the major players in EMT, was not detectable in the cells missing the AKT1 isoform. Results indicate an association of Twist1 with AKT1 in EMT and migration of ovarian tumors cells. This finding is significant because AKT2 has been implicated as the major player of cell migration in human breast cancer iv cells. Collectively, these findings support a tissue specific role of the AKT isoforms, and may provide insights regarding the most useful cell context in order to target components of the AKT signaling pathway indirectly affecting EMT in order to prevent tumor progression in patients with ovarian and perhaps other types of cancers

Yes / HOX genes are vital for all aspects of mammalian growth and differentiation, and their dysregulated expression is related to ovarian carcinogenesis. The aim of the current study was to establish the prognostic value of HOX dysregulation as well as its role in platinum resistance. The potential to target HOX proteins through the HOX/PBX interaction was also explored in the con-text of platinum resistance. HOX gene expression was determined in ovarian cancer cell lines and primary EOCs by QPCR, and compared to expression in normal ovarian epithelium and fallopian tube tissue samples. Statistical analysis included one-way ANOVA and t-tests, using statistical software R and GraphPad. The analysis identified 36 of the 39 HOX genes as being overex-pressed in high grade serous EOC compared to normal tissue. We detected a molecular HOX gene-signature that predicted poor outcome. Overexpression of HOXB4 and HOXB9 was identified in high grade serous cell lines after platinum resistance developed. Targeting the HOX/PBX dimer with the HXR9 peptide enhanced the cytotoxicity of cisplatin in platinum-resistant ovarian cancer. In conclusion, this study has shown the HOX genes are highly dysregulated in ovarian cancer with high expression of HOXA13, B6, C13, D1 and D13 being predictive of poor clinical outcome. Targeting the HOX/PBX dimer in platinum–resistant cancer represents a potentially new therapeutic option that should be further developed and tested in clinical trials. / This research was supported by GRACE, a gynaecological charity based in Surrey, UK.