Endometrial cancer is the most common female reproductive cancer in the United States. Most known risk factors for endometrial cancer are either genetic or related to exposure to estrogens; less is known about risk due to environmental exposures. While several studies have examined the relationship between perineal powder use and ovarian cancer risk, only one study has addressed the relationship with endometrial cancer risk. The Women's Health Initiative Observational Study, a prospective cohort study of 93,676 United States postmenopausal women from 1993-2005, measured perineal powder use at baseline via self-report. Cases of endometrial cancer were self-reported and confirmed by both local and central physician adjudicators. Cox proportional hazards regression was used to examine the association between perineal powder use and endometrial cancer, adjusting for known risk factors. Of the 48,912 women in our analysis, 25,181 (52%) reported ever use of perineal powders. There were 452 incident cases of endometrial cancer diagnosed during 366,872 person-years of follow-up. Ever use of perineal powder was not significantly associated with increased risk of endometrial cancer (hazard ratio 1.05, 95% confidence interval 0.87-1.27). However, use of any perineal powder for 20 or more years was associated with a 30% increase in risk (hazard ratio 1.30, 95% CI 1.01-1.67) compared to never users. Use of powder on both a diaphragm and the perineal area was associated with a 39% increase in risk (hazard ratio 1.39, 95% CI 1.00-1.93). Cessation of perineal powder use, particularly on a diaphragm, may help reduce risk of endometrial cancer.

京都大学 / 新制・課程博士 / 博士(医学) / 甲第23778号 / 医博第4824号 / 新制||医||1057(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 黒田 知宏, 教授 溝脇 尚志, 教授 波多野 悦朗 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM

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A cell’s interaction with its extracellular environment is critical to tissue structure and function. This work investigates the effect of substrate topography on selective cell adhesion and morphology. Alterations in cell response to micro- and nanoscale signals and cues can cause changes in downstream functions of proteins and complexes such as invasive and metastatic motility of malignant tumour cells and the differentiation direction of stem cells. Biomaterial surfaces can be modified to provide different chemical and topographical cues and encourage controlled cell-substrate interaction. At the protein level, template substrates have shown and increased affinity for selective binding of the imprinted antigen or antibody. Topography of a cell’s microenvironment may be replicated as a permanent polymer mould by bioimprinting technology, which was developed at University of Canterbury. The resulting high resolution methacrylate polymer samples have been used for imaging and analysis, but have not previously been investigated as cell culture substrates. This work investigates the effect of bioimprint and photolithographic substrate patterning on cell behaviour in culture. Optimisation of a methacrylate co-polymer resulted in a 6:3:1 ethylene glycol dimethacrylate: methacrylic acid: photoinitiator polymer mixture cured by 240 seconds of UV exposure. The polymer was used to replicate cell membrane features into a permanent polymer mould [a bioimprint]. The resulting high resolution methacrylate bioimprints were cleaned and sterilised for use as a secondary cell culture substrate. Ishikawa endometrial cancer cells were cultured on bioimprinted methacrylate polymer substrates. Preliminary results showed preferential cell adhesion to bioimprinted areas over flat areas and also showed three dimensional spheroid growth instead of lateral two dimensional monolayer spreading. At higher seeding densities, preferential adhesion was similarly noted as well as peeling artefacts of shear stresses and cell size variation on flat methacrylate substrate regions. Fluorescent imaging and cell culture stencilling highlighted the association of secondary cells with bioimprint substrate features. To determine whether preferential cell adhesion effects were due to bioimprint features or general topography modification, secondary cancer cells were cultured on comparable photolithographically-defined, geometrically-patterned substrates. Methods for transferring regular pattern arrays into methacrylate polymer substrates were developed. No organisation or preferential adhesion effects were observed in association with pillar and hole patterns between 5-30 µm. However, artefact incidence in methacrylate polymer replication techniques led to development and adaptation of polystyrene patterning techniques. Experimental analysis of substrate-dependent effects on cell culture adhesion and organisation was extended to a non-cancerous cell line model. C2C12 mouse skeletal muscle cells were chosen for these investigations because of their ability to differentiate further, into myocytes or myofibrils. C2C12 myoblasts seeded on common cell culture substrates showed a notable morphology variation and extent of differentiation between cells grown on tissue culture polystyrene [TCPS] and polydimethylsiloxane [PDMS]. Myoblasts were plated on geometrically-patterned polystyrene and PDMS substrates. Significant alignment to grated pattern features was observed on both substrate types, before and after driven differentiation. Peeling artefacts of confluent tissue-like culture from PDMS surfaces which were observed were unreported previously in literature. The results reported in this thesis provide a foundation for potential research and commercial application for surface modification methods. The biomimetic topography provided by bioimprinted substrates can be used to identify and investigate cell activities, including for example the mechanisms of cell adhesion and separation in metastatic and invasive cancer research. Altering the material of the bioimprinted substrates may attune substrate topographies as scaffolds to direct specific stem cell differentiation for regenerative tissue engineering applications.

Endometrial cancer (EC) is the fourth most common cancer affecting women in the UK. Those with a family history of EC have an increased risk compared with the general population. Highly penetrant germline mutations in mismatch repair (MMR) genes and DNA polymerases account for only a small proportion of the familial aggregation. The aim of this thesis is to investigate the genetic susceptibility to EC in the general population using cases and controls of European ancestry. A GWAS meta-analysis totalling 7,737 EC cases and 37,144 controls yielded five novel EC risk loci of genome-wide significance (P < 5x10⁻⁸). In decreasing order of significance, these were at chromosomes 13q22.1 (rs11841589, near KLF5), 6q22.31 (rs13328298, in LOC643623 and near HEY2 and NCOA7), 8q24.21 (rs4733613, telomeric to MYC), 15q15.1 (rs937213, in EIF2AK4, near BMF) and 14q32.33 (rs2498796, in AKT1, near SIVA1). A second independent EC signal was found in the 8q24 locus. The association found in a previous EC GWAS at HNF1B on chromosome 17 was replicated at a higher significance, with the most significant SNP being rs11263763. CYP19A1 SNPs have previously been associated with EC and higher circulating levels of oestrogen from candidate studies, but I confirmed this locus to be genome-wide significant for the first time. Functional annotation and in vitro studies for the EC risk loci at the intergenic region of chromosome 13q22 suggested that the functional SNP sits within a transcriptional repressor for KLF5, with the higher-risk allele reducing repressor activity. The propensity for germline MMR and DNA polymerase muations to cause both EC and colorectal cancer (CRC) prompted me to search for common variants associated with both cancer phenotypes. An EC CRC GWAS meta-analysis showed little evidence of shared susceptibility loci. However, this meta-analysis revealed a novel genome-wide significant risk locus: rs3184504, a missense SH2B3 SNP that has not previously been associated with either EC or CRC. This thesis has enhanced the understanding of genetic susceptibility to sporadic EC and increased the number of genome-wide EC-associated variants to seven.

Mutations in FGFR2 are common in a subset of endometrial carcinomas. Given the emergence of small molecule inhibitors specific to this receptor tyrosine kinase, FGFR2 is an attractive therapeutic target. However, compensatory and adaptation mechanisms limit the clinical utility of compounds that target nodes in the receptor tyrosine kinase network. Here, we analysed the impact of FGFR inhibition in endometrial cancer cells and observed the emergence of a resistant population in an FGFR2-mutant cell line. To understand the mechanisms underlying this adaptation response, we used a phosphoproteomics approach to measure the kinase network in an unbiased manner. These experiments led to the identification of an AKT-related compensatory mechanism underpinning this resistance. Further dissection of this resistance mechanism utilising gene expression analysis showed PHLDA1, a negative regulator of AKT, was significantly down-regulated in resistant cells. This was further confirmed at the protein level. siRNA knockdown of PHLDA1 conferred immediate drug resistance in the FGFR2-mutant endometrial cancer cell line. Therefore, we identified PHLDA1 down-regulation as a mediator of drug resistance in FGFR2 mutant cancer cells, the first demonstration of the role of PHLDA1 in the acquisition and maintenance of drug resistance. Using a 3D physiomimetic model, we demonstrated that AKT inhibition alone also led to generation of a drug-resistant population. Most importantly, dual-drug therapy inhibited proliferation and induced cell death. Our data highlight how mass spectrometry and microarray gene expression analysis can complement each other in the identification of novel resistance mechanisms in cancer cells. These data suggest that combination treatment of FGFR2-mutant endometrial cancers, targeting both FGFR2 and AKT, represents a promising therapeutic approach.

El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / Background Obesity, insulin resistance and type 2 diabetes mellitus (T2DM) have been associated with endometrial cancer (EC). In this systematic review and meta-analysis we evaluated the effect of metformin on clinical outcomes in patients with EC and insulin resistance or T2DM. Methods Four research databases were searched for original articles published in all languages up to 30 October 2016. Outcomes of interest were overall mortality (OM), cancer-specific mortality, disease progression, and metastases. We performed a random effect meta-analysis of adjusted effects expressed as hazard ratios (HR); heterogeneity among studies was described with the I2 statistic. Results Of the 290 retrieved citations, 6 retrospective cohort studies in women with EC (n = 4723) met the inclusion criteria, and 8.9% to 23.8% were treated with metformin; OM data was available from 5 studies. In 4 studies of EC patients (n = 4132), metformin use was associated with a significant reduction in OM in comparison with not using metformin (adjusted HR [aHR] 0.64, 95% CI 0.45–0.89, p = 0.009). In three studies evaluating patients with EC and T2DM (n = 2637), metformin use was associated with a significant reduction in OM (aHR 0.50, 95%CI 0.34–0.74, p = 0.0006). There was low to moderate heterogeneity of adjusted effects across studies. There was no information about the effect of metformin on cancer-specific mortality, disease progression, or metastases. Conclusions Metformin treatment is associated with a significant reduction in OM irrespective of diabetes status in patients with EC. The survival benefit suggests that diabetes screening and maintenance of good glycemic control may improve outcomes in EC. / Revisión por pares

Introduction: Over 74,000 US women are diagnosed annually with endometrial or cervical cancers and experience significant treatment-related difficulties with body image, sexuality, and sexuality. The aims of the research were to examine relationships among body image, sexuality, and sexual functioning, and understand women’s views of these concepts. A novel theoretical framework encompassing these three concepts and contextual variables guided the research. Methods: A non-experimental cross-sectional pilot study was conducted with pre-menopausal women in an academic medical center who had stages I-III cervical or endometrial cancer, were 3-36 months post-treatment, and had no mental health diagnoses. Participants completed the Body Image Scale, Female Sexual Function Index, Female Sexuality Questionnaire, a demographic questionnaire, and open-ended questions and the researchers collected clinical data from the medical record. Results: Twenty women participated, and most reported disruption in body image, sexuality, and sexual functioning. Significant relationships were found between body image and sexuality (p = 0.0244) and sexuality and sexual functioning (p <0.0003). Stages II-III disease were significantly (p= 0.0371) associated with worse body image. Women reported issues with body image, sexuality, sexual functioning, psychosocial areas, reproduction, and communication with healthcare providers and personal caregivers. Discussion: This study is one of the first to examine relationships among body image, sexuality, and sexual functioning simultaneously, highlighting the importance of this approach. Despite design and sample limitations, results demonstrate a need for longitudinal studies using larger samples to further examine relationships among these concepts and selected contextual variables, explore communication issues, and refine the theoretical framework.

Cancer is the second leading cause of death in the USA, following cardiovascular disease. Treating cancer using conventional therapies is associated with low response rates and high toxicity, because these therapies usually lack specific tumor accumulation. Loading anticancer drugs into intelligently designed polymeric nanoparticles (NPs) can serve in delivering these drugs specifically to the tumor site, thus boosting their efficacy and reducing any associated off target toxicity. Targeting NPs to the tumor site can occur through either passive or active means. In passive targeting, NPs of specific size and surface characteristics can exploit the tumor’s erratic vasculature and occluded lymphatic drainage to extravasate the systemic circulation and accumulate preferentially at the tumor site. Active targeting mandates grafting the surface of NPs with a ligand that specifically interacts with a protein expressed at higher levels at the tumor site, in comparison to elsewhere in the body. In the current research, we independently investigated the utilization of passive and active targeting strategies to treat aggressive forms of cancer. Initially, passively targeted poly(lactic-co-glycolic acid) (PLGA) NPs to treat aggressive forms of endometrial cancer (EC) were investigated. A novel combination of soluble paclitaxel (PTX), a first line chemotherapy for EC, and soluble BIBF1120 (BIBF, nintedanib), an antiangiogenic molecular inhibitor, was first tested against three EC cell lines bearing different p53 mutations. The results showed that only EC cells with loss of function (LOF) p53 were sensitive to the combination therapy, indicating the potential of this combination to engender synthetic lethality to PTX. Next, NPs loaded with PTX were investigated with respect to the impact of varying the polymer lactic acid to glycolic acid ratio and the surfactant type on the major physicochemical properties of the prepared nanoparticles, drug loading, cellular uptake, cytotoxicity, and drug release. The optimum formulation was then loaded with BIBF and the combination of independently loaded passively targeted NPs were further evaluated for in vivo activity against a xenograft model of LOF p53 EC. The combination of independently loaded NPs exhibited the highest reduction in tumor volume and prolonged survival when compared to soluble PTX, PTX NPs or untreated control. These data highlight this specific combination of NPs as a novel promising therapy for LOF p53 EC. In a second study, the use of actively targeted NPs to treat liver cancer was explored. In this study, a combination of small interfering RNA (siRNA) against astrocyte elevated gene-1 (AEG-1), and all-trans retinoic acid (ATRA) was investigated as a new therapy for hepatocellular carcinoma (HCC). AEG-1 is a highly expressed oncogene that is directly involved in HCC progression and aggressiveness, in addition to reducing the ability of retinoic acid to induce apoptosis in HCC cells. First, a new conjugate was synthesized that was capable of delivering siRNA selectively to HCC cells, using galactose as a targeting moiety. The conjugate was prepared by linking poly(amidoamine) (PAMAM) dendrimers, polyethylene glycol (PEG) and lactobionic acid (Gal, disaccharide containing galactose) (PAMAM-PEG-Gal). We confirmed the synthesis of the conjugate using 1H-NMR, Mass spectrometry and Matrix-Assisted Laser Desorption/Ionization (MALDI) Mass Spectrometry. Next, nanoplexes of the synthesized conjugate, PAMAM-PEG-Gal, and AEG-1 siRNA were prepared. Nanoplexes were further characterized for their size, surface charge, morphology, and electrophoretic mobility to identify the optimum complexation ratio between PAMAM-PEG-Gal and the siRNA. Then, mice bearing orthotopic luciferase expressing HCC cells were treated with the optimum nanoplex formulation. Results showed that a combination of AEG-1 nanoplexes and ATRA results in a significant reduction in luciferase expression, reduced liver weight, lower AEG-1 mRNA levels and increased apoptosis, when compared to utilizing nanoplexes with silencing control (siCon), siCon+ATRA, or AEG-1 nanoplexes alone. The results indicate that the combination of liver-targeted AEG-1 nanoplexes and ATRA may be a potential treatment for aggressive HCC. These data place targeted NPs as a promising efficient delivery system for cancer treatment.

Individuals who identify themselves as transgender have gender identities that do not match their anatomical sex. Females who identify as male, also known as female-to-male transgender (FTM), may opt to undergo hormonal and surgical treatment in order to transition to the male phenotype, including high-dose testosterone treatment to develop male secondary sexual characteristics and surgical procedures. Currently, the recommendation is for the patient to have a hysterectomy within five years of initiating testosterone therapy to decrease the risk of developing endometrial cancer. However, long-term testosterone treatment has not been proven to cause an increased risk of endometrial cancer. With the use of gene expression and immunohistochemical studies, this study aimed to show no upregulation of genes associated with proliferation (Ki-67) and endometrial cancer (ZIC2) in endometrial tissue from FTM individuals treated with long-term testosterone compared to endometrial tissue from postmenopausal women, premenopausal women with benign endometrium, and women with endometrial cancer. Our findings showed that Ki-67 and ZIC2 expression in the FTM samples was significantly lower than in the endometrial cancer samples. Our findings call into question the concept that long-term testosterone treatment causes neoplastic changes in endometrial tissue and the need for routine hysterectomy in these patients. / 2018-07-11T00:00:00Z