Abstract Introduction Knowledge about the oncologic status of pelvic lymph nodes forms an essential and integral part in the management of women with uterine cancer. Lymph node status is part of endometrial cancer staging and plays an important role in primary treatment and adjuvant treatment planning and prognosis in women with cervical cancer. Current practice in the management of uterine cancers involves systematic full pelvic lymphadenectomy, mainly to determine the oncological status of the nodes, as there is no high-quality evidence suggesting a therapeutic effect attributable to lymphadenectomy. Imaging in the form of computed tomography (CT) scans and magnetic resonance (MRI) scan is not accurate to determine pelvic lymph node status in women with uterine cancer. Functional scans such as 18Fluoro-deoxy-glucose positron emission/computed tomography (FDG-PET/CT) scan might provide better access in this setting. Sentinel lymph node biopsy (SLNB) procedures, specifically the SLNB algorithm, have been proposed as a safe and accurate alternative procedure to full systematic lymphadenectomy in women with uterine cancers. It has also been proposed as a better alternative than complete omission of lymphadenectomy in women with presumed low risk early stage endometrial cancer. SLNB procedures might also be able to detect higher rates of lymph node metastases with the detection of micro metastases following pathological ultrastaging The presence or absence of high risk human papilloma virus (hrHPV) DNA in sentinel lymph nodes of women with cervical cancer has also been suggested to be a useful adjunct to frozen section examination (FSE) in assisting with determination of the status of the non-sentinel nodes. Some data suggest the combination of negative FSE and absence of hrHPV accurately predict the absence of metastases. South African women have high prevalence of human immunodeficiency virus infection, tuberculosis (TB) and pelvic inflammatory disease (PID). All these infections involve the lymphatic system. Data on SLNB procedures are form well-developed countries with different disease burdens and socioeconomic profiles, and there is no data from women living in low-resource settings. Aims This study aimed to determine the efficacy of and performance of FDG-PET/CT scan and SLNB and SLNB algorithm in accurately predicting the regional lymph node status of the pelvis in women with early stage cervical cancer and presumed early stage endometrial cancer. It also aimed to investigate the usefulness of HPV DNA testing of sentinel nodes in women with cervical cancer. Population and setting This was a prospective observational study performed in the Gynaecologic Oncology Unit at the Kalafong Provincial Tertiary Hospital and Steve Biko Academic Hospital. Patients aged 18 years and older, with operable stages cervical cancer and presumed early stage endometrial cancer willing and able to provide informed consent were eligible for inclusion. Materials and methods Sentinel node mapping was done using methylene blue (MB) and indocyanine green (ICG) injected into the cervix after induction of anaesthesia at the time of primary surgery. 99Technetium nanocolloid (99Tc) was administered one day pre-operatively followed by lymphoscintigram. FDG-PET/CT scans were performed prior to surgery. Following mapping and removal, FSE, HPV DNA typing, haematoxylin and eosin (H&E) examination with ultrastaging on H&E negative specimens were performed on the SLNs. All patients underwent systematic full pelvic lymphadenectomy and appropriate cancer surgery. Results One hundred patients were prospectively recruited to the study and results of 94 patients were available for analysis. SNL detection rate of the whole group was 60.6% with bilateral detection 29.2%. Twenty-four patients (25.5%) had pelvic metastases. Sixty-five percent of women with cervical cancer in this study were HIV positive, and the SLN detection rate in this group was 65% with bilateral detection rate of 30%. The detection rate was significantly higher in women without nodal metastases, those with stage IA2 – IB2 disease, with tumour less than 2 cm and women with BMI less than 25 kg/m2. HIV status, history of TB, PID and the presence of adhesions did not influence the SLN detection rate. The sentinel lymph node biopsy algorithm has a sensitivity of 100%, NPV of 100% and a false negative rate of 0% in this study. The SLNB procedure identified two women with only micro metastases (15.4%). These women would not have been identified with systematic lymphadenectomy and H&E examination. Indocyanine green and the combination of methylene blue and 99Technetium nanocolloid had significantly better sentinel node detection rates compared to methylene blue alone FDG-PET/CT scan was performed in 28 women. The sensitivity, specificity, positive and negative predictive values of FDG-PET/CT scans to accurately predict nodal status, were 66.67%, 82%, 30.77% and 95.38% respectively. The false negative rate of FDG-PET/CT scans was 33.3%. The sensitivity, specificity, PPV and NPV for FSE in this cohort was 66.67%, 100%, 100% and 96.05% respectively. The FNR for FSE was 23.1%. Thirty-two patients with cervical cancer had tumour and SLN hrHPV DNA data. The sensitivity, specificity, PPV and NPV of sentinel lymph node HPV DNA to predict metastases was 50%, 69.6%, 30 and 84.2% respectively with a false negative rate of 42.8%. Conclusions Although the SLN detection rate was lower compared to the published literature, the SLNB algorithm performed excellently in this group of patients of which the majority were HIV-infected. The SLNB procedure can be considered as a treatment option in selected cases in the management of women with early stage endometrial and cervical cancer. PET/CT should not be used as part of the primary diagnosis and staging investigations in women with uterine cancer, and is recommended only in selected cases for initial staging of locally advanced cervical cancer being considered for radical chemoradiation therapy. In this study, testing for the presence of hrHPV DNA in the sentinel lymph nodes was not useful as a predictor of pelvic lymph node status. The combination of negative FSE and negative hrHPV in the SLNs did not have a reliable negative predictive value for the absence of pelvic nodal metastases. / Thesis (PhD)--University of Pretoria, 2017. / Obstetrics and Gynaecology / PhD / Unrestricted

Tumors are mutated abnormal groups of cells that develop at any stage of life in any part of the body. Mammilla tumors appear in chest tissue that contain malignant cells in the terminal ductal-lobular unit, where the risk of the development of a mammilla tumor increases with age with a probability of 14.7%. Previous reviews have only focused on radiotherapy and digital mammography, while this review is, to the best of the author´s knowledge, the first review that encompasses the tomosynthesis and presumptive magnetic resonance using digital mammography. The aim of the meta-analysis was to determine the extent in which mass-testing of mammilla tumor metastasis can lead to its mitigation in adult females of all age-groups. The research question was the following: To what extent can mammilla tumor metastasis be mitigated by mass-testing of adult females of all age-groups? As part of the meta-analysis, a literature review was conducted using a selection of keywords in search queries on Pubmed, Libsearch and Academic Search Elite. In conclusion, mass-testing of mammilla tumor metastasis does not lead to a mitigation in adult females of all age-groups, since there was not a statistical significance of pooled value as indicated by the forest plot and the funnel plot indicated that the publication bias had some effect and the Mann-Whitney U-test also indicated that there was not a significance difference. Future research may consist of whether adult females within the age-range of 60-80 benefit from the test.

Thesis (MS)--Boston University / PLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you. / Sexuality is one quality of being human. Sexual health must be addressed as a significant and integral element of total health care. Nurses caring for patients with oncology and neuromuscular related disabilities are presented with situations concerning sexual dysfunctions that require judgement, knowledge and sensitivity. Yet, studies have shown that nurses demonstrate low sexual knowledge levels and conservative sexual attitudes. They often neglect this aspect of patient care. The sexual attitudes of nurses may be a significant obstacle to their effective functioning in the field of sexual counseling. This proposed study will investigate sexual attitudes of oncology nurses and rehabilitation nurses. / 2031-01-01

Neuroendocrine tumors (NETs) are a rare type of malignancy, however, their incidence and prevalence have markedly increased over the past several decades. Gastroenteropancreatic neuroendocrine tumors (GEP-NETs), NETs of the gastrointestinal (GI) tract, account for the majority of these tumors. Data from the Surveillance, Epidemiology, and End Results (SEER) program of the National Cancer Institute (NCI), as well as from multiple cancer registries in Canada and Europe, indicate that the small intestine is one of the most common primary sites of GEP-NET development in Caucasian populations. Smaller studies from Asian countries, however, report a significantly lower proportion of small intestinal NETs (SI-NETs) in those countries’ populations. Interestingly, this finding persists in a study of Asians living in the United States, which, therefore, may suggest that genetics, and not environment, play a predominant role in driving this difference. Multiple studies exploring the genetics of SI-NETs in Caucasians have been published and have identified several genetic alterations implicated in tumorigenesis. However, the most consistently reported finding is chromosome 18 loss of heterozygosity (chr18 LOH), which has been detected in 61-78% of SI-NETs. Unfortunately, these studies have yet to be performed in a strictly Asian cohort. The proposed study, therefore, is a multicenter, cross-sectional study that will compare the proportion of SI-NETs with chr18 LOH in Asian versus Caucasian populations by performing a fluorescence in-situ hybridization (FISH) analysis of SI-NET tissue from both Asian and Caucasian samples. Overall, the results of this study may help to identify possible mechanisms driving the difference in proportion of SI-NETs between Asians and Caucasians, which in turn, may help to better characterize the genetic events leading into SI-NET tumorigenesis.

The purpose of this paper is to characterize the glucocorticoid receptor (GR) signaling and relevance in the context of enzalutamide resistant prostate cancer cells. Enzalutamide is a drug that functions to dampen androgen receptor (AR) signaling, thus inhibiting cancer dependency on the receptor protein. Although the application of the drug reduces AR signaling in these cancer cells, an alternate pathway involving GR signaling may be upregulated as a compensatory bypass mechanism. Therefore, it possible that GR assumes the role of AR and facilitates tumor growth by promoting the expression of genes regulated by AR. To analyze how GR operates, we analyzed GR signaling in enzalutamide resistant metastatic prostate cancer cell lines. We assessed protein levels of AR and GR as well as mRNA expression of various AR targets. Our results illustrate the expected downregulation of AR and upregulation of GR in enzalutamide resistant cells. Furthermore, some canonical AR targets like prostate specific antigen (PSA), Prostate Specific Membrane Antigen (PSMA) and Prostatic Acid Phosphatase (PAP) were inhibited by a novel GR inhibitor. Thus, this GR inhibitor could be used in combination with enzalutamide and create a more potent AR signaling blockade. Prostate cancer is a very problematic disease in men and becomes especially challenging to treat during the metastatic stage as they are non-sensitive to anti-androgens. The significance of understanding how GR functions, as well as the potential benefit of blocking GR signaling, may provide insight into novel drugs and agents that could specifically target these pathways, control and mitigate cancer growth, and prolong the lives of patients.

Osteosarcoma is a rare osteogenic cancer which is a predominantly pediatric/young adult disease but has a large adult population. Due to the skewed patient population, randomized clinical trials have focused on the younger osteosarcoma cases, and then applying these treatments to adults. Yet these treatments have not been tested in a prospective randomized clinical trial for adults for whether they achieve the same outcomes as those in the pediatric population. The purpose of this study is to compare outcomes of pediatric and adult osteosarcoma cases through a retrospective review of patients treated at two facilities, Dana-Farber Cancer Institute (DFCI) and Massachusetts General Hospital (MGH). A list of 2,263 potential patients was provided by the pathology departments of DFCI/MGH. Eligibility required: 1) the patient to be at least 18 YO at the time of diagnosis, 2) a pathology report confirming osteosarcoma, 3) the available follow-up data is greater than 1 year (unless the patient died within that year). The electronic charts were assessed for prognostic variables which includes; date of diagnosis, age at diagnosis, sex, primary disease site, tumor size, histologic response, and treatment course. The overall survival and metastasis free survival were based on the date of pathologic confirmation of osteosarcoma. The collected data was analyzed through the Kaplan-Meier method to produce overall survival and metastasis free survival curves. The 5-year OS and MFS rates were calculated with those alive at last follow-up censored from the analysis. The prognostic variables were tested for significant difference in OS and MFS by ANOVA and t-tests. Of potential patients, 177 were deemed eligible for this study and were found to have 5-year MFS and OS of 45.8% and 64.4%, respectively. When compared against results from a primarily pediatric/young adult study, our patient population had a significantly lower OS and MFS. This finding indicates that current treatment regimens have a shorter effect in adult osteosarcoma cases when compared to pediatric patients. Although pediatric patients have a sustained response to current treatment therapies, the adult cases of osteosarcoma have a significantly shortened outcome which needs to be addressed in future research.

Gastro-esophageal adenocarcinomas (GEA) are among the fastest rising malignancies in North America. Despite advances in cancer prevention and treatment for other cancers, the number of GEA cases continues to rise and prognosis remains bleak with five-year survival rates of only 20%. Additionally, many GEA patients won’t respond to first line therapy, many may develop therapeutic resistance, or will show disease recurrence. Previous drug screen models failed clinical trials due to the failure of the model to adequately recapitulate the primary sample. A new model, the patient-derived organoid (PDO), has become the newest method of investigating and testing numerous characteristics of the in vivo tumor. Initial studies have demonstrated the organoid’s advantages: PDOs are highly heterogeneous, may be maintained in culture indefinitely, and have the capability to model carcinogenesis and therapeutic response. However, limitations exist and questions remain that have yet to be addressed. Indeed, one of the challenges of using organoids is knowing whether the organoids are recapitulating normal or tumor tissue. Additionally, there seem to be limits on immortality of the organoids and the heterogeneity. Finally, without the stroma and Tumor Microenvironment (TME) in culture, the model is limited in its ability to test the response to immunotherapy-based drugs. Current research aims to develop a clinical pipeline utilizing organoids regularly as a diagnostic tool to evaluate therapeutic response, identify emergence of chemoresistance and perform targeted drug screens. Overall, PDOs are a burgeoning method of investigating GEA and are a powerful translational tool from bench to bedside.

OBJECTIVES: To determine the effect of murine cytomegalovirus (MCMV) infection on the growth and proliferation of glioblastoma using murine models of differing genetic backgrounds, as well as to determine whether the observed effects are dependent on the mutational background of the glioblastoma models. METHODS: An experimental orthotopic murine glioblastoma model was established using latently MCMV infected C57BL/6 mice, with non-infected mice as controls. Mouse brains were harvested at the end-point and subject to histological analysis using immunostaining and visualization by confocal microscopy. Viral infectivity and protein expression levels were assayed using time-lapse microscopy and western blotting. RESULTS: Our results indicate that while the degree of viral infection may differ between experimental cell lines, MCMV-infected tumor-bearing mice consistently demonstrate accelerated tumor growth and worsened survival relative to the uninfected controls. Additionally, our results provide further evidence to support the previously reported findings implicating CMV as a driver of angiogenesis in vivo, as sections from MCMV-infected mice showed a greater density of blood vessels and more complete blood-brain barrier formation. Finally, our results show that the addition of antiviral agents as an adjunct therapy to first-line chemotherapeutics effectively reduced tumor sphere size and partially reversed viral infection by MCMV. CONCLUSIONS: Here we demonstrate the tumor-promoting effects of MCMV infection are consistently observed in different glioblastoma mouse models suggesting that these effects are independent of the tumor genetic background of our tested tumor models. Moreover, the success of preliminary in vitro studies using a combination treatment of antiviral and chemotherapy strengthen the case for targeting CMV therapeutically in the treatment of GBM.

Extracellular vesicles (EVs) include exosomes and microvesicles, and are important mediators of cell-to-cell communication in healthy and pathological environments. Packed with biological information and present in a variety of biological fluids, EVs offer exciting promise for biomarker discovery and applications in therapeutics and non-invasive diagnostics. Despite their potential medical use, many of the mechanisms underlying EV biogenesis, cargo packaging, and secretion remain unknown. To an extent, these findings have been restricted by the limited cells from which vesicles have been characterized utilizing the same enrichment method. The following work contributes to advancement of knowledge surrounding EV formation within the cell, protein trafficking into EVs, and the content and functions of oncogenic vesicle secretion in cancer. In Chapter 2 of this dissertation, vesicle secretion is characterized across sixty cancer cell lines from the National Cancer Institute (NCI-60) by nanoparticle tracking analysis. The quantity of EVs secreted by each cell line was compared to reference transcriptomics data to identify gene products associated with vesicle secretion. Positive correlates of exosomal-sized vesicle secretion included Rab GTPases and mediators of sphingolipid metabolism, while larger microvesicle-sized vesicle secretion was associated with gene products involved in cytoskeletal dynamics and exocytosis, as well as Rab GTPase activation. One of the identified targets, CD63, was further evaluated for its role in vesicle secretion. Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) and CRISPR-associated protein 9 (Cas9) knockout of the CD63 gene in HEK293 cells resulted in a decrease in small vesicle secretion, suggesting the importance of CD63 in exosome biogenesis. To investigate the global protein content of NCI-60 cell derived EVs, pure vesicle isolates were harvested for a large-scale mass spectrometry approach, detailed in Chapter 3. This work provides the largest proteomic profile of EVs in a single study, identifying 6,071 proteins with 213 common to all isolates. Proteins included established EV markers, and vesicular trafficking proteins such as Rab GTPases and tetraspanins. The identification of differentially-expressed proteins may offer future candidates for earlier cancer diagnosis and prognostic monitoring of disease. Network analysis of vesicle quantity and proteomes identified EV components associated with vesicle secretion, including CD63, CD81, syntenin-1, VAMP3, Rab GTPases, and integrins. Finally, integration of vesicle proteomes with whole-cell molecular profiles revealed similarities, suggesting EVs provide a reliable reflection of their progenitor cell content, and are therefore excellent indicators of disease. Together, these observations reveal new insights into key players involved in exosome and microvesicle formation, and may provide a means to distinguish EV sub-populations. In both the aforementioned works, a tetraspanin protein, CD63 was identified and confirmed for its role in small vesicle biogenesis. CD63 is a conserved protein enriched in late endosomal and lysosomal compartments and has been widely used as an exosome marker. However, little is understood about the mechanisms of CD63-mediated exosome formation, or its interacting proteins. Accumulating evidence has suggested that an Epstein-Barr virus-encoded oncoprotein, latent membrane protein 1 (LMP1) may interact with CD63 within the exosomal pathway. Viruses share similar sizes and structures as exosomes, and recent evidence suggests that viruses can hijack the exosome pathway to modulate cell-to-cell signaling. Indeed, LMP1 trafficking into multivesicular bodies (MVBs) can alter the content and function of exosomes. LMP1-modified exosomes can in turn enhance the growth, migration, and invasion of malignant cells, demonstrating the capacity to manipulate the tumor microenvironment and enhance the progression of EBV-associated cancers. Despite the growing evidence surrounding the significance of LMP1-modified exosomes in cancer, very little is understood about the mechanisms that orchestrate LMP1 incorporation into these vesicles. In efforts to advance knowledge surrounding the role of CD63 in exosome production and vesicular protein sorting, original work in Chapter 4 demonstrates the importance of CD63 presence for exosomal packaging of LMP1. In this study, a role of LMP1 in vesicle production that requires CD63 is described, providing an extensive demonstration of CD63-mediated exosomal LMP1 release that is distinct from lipid raft trafficking. Further evidence revealed the role of CD63 in limiting LMP1-induced non-canonical NF-κB and ERK activation. Overall, these findings have implications in future investigations of physiological and pathological mechanisms of exosome biogenesis, protein trafficking, and signal transduction, especially in the context of viral- or nonviral- associated tumorigenesis. / A Dissertation submitted to the Department of Biomedical Sciences in partial fulfillment of the Doctor of Philosophy. / Spring Semester 2017. / February 17, 2017. / Biomarkers, Cancer, Epstein-Barr virus, Exosomes, Extracellular vesicles, Proteomics / Includes bibliographical references. / David G. Meckes, Jr., Professor Directing Dissertation; Scott M. Stagg, University Representative; Pradeep G. Bhide, Committee Member; Myra M. Hurt, Committee Member; James M. Olcese, Committee Member.

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