Orientadores: Liliana Aparecida Lucci De Angelo Andrade, Sophie Francoise Mauricette Derchain / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-24T12:06:20Z (GMT). No. of bitstreams: 1 Pinto_PaolaBertolottiCardoso_D.pdf: 22982855 bytes, checksum: ca4a4e679f0037c1494bf2866a333968 (MD5) Previous issue date: 2013 / Resumo: Os carcinomas mucinosos do ovário são raros e representam apenas 3% dos carcinomas. Frente a este diagnóstico, é preciso descartar a possibilidade de metástase para o ovário, principalmente de neoplasia primária do trato gastrointestinal. Apesar da avaliação morfológica, macro e microscópica, e das reações imunoistoquímicas contribuírem para o diagnóstico diferencial, existem casos de difícil diferenciação. Um algoritmo para separar os carcinomas mucinosos primários dos metastáticos no ovário foi proposto na literatura e determina que são metastáticos os tumores bilaterais ou unilaterais menores que 13cm, classificando as neoplasias com uma acurácia de quase 90%. Objetivos: comparar os aspectos macro e microscópicos aliados à avaliação imunoistoquímica para a diferenciação entre tumores mucinosos primários e metastáticos no ovário, avaliando a acurácia do algoritmo nos casos, os dados clínicos e sua evolução. Métodos: Todos os tumores mucinosos envolvendo o ovário, dos arquivos do Laboratório de Anatomia Patológica da UNICAMP no período de 1994 a 2009 foram levantados. Feita revisão dos prontuários com descrição dos dados clínicos, evolução das pacientes, revisão de lâminas para avaliação de dados histopatológicos e seleção dos blocos de parafina para a construção de micro-arranjo de tecidos, onde foram realizadas as reações imunoistoquímicas para: CK7, CK20, ?-catenina, WT-1, CDX-2, Dpc-4, CA125, RE e RP. Resultados: Dos 76 casos selecionados, 35 eram carcinomas mucinosos primários do ovário, 33 eram metastáticos e em 8 casos o primário não foi definido, sendo excluídos da análise estatística. A sobrevida média foi maior nos primários (65X35 meses; p<0.0001). A acurácia do algoritmo foi de 82,1%. A maioria dos metastáticos originou-se do cólon ou reto (54%). Dos primários, 85% eram unilaterais >13 cm e dos metastáticos, 61% eram bilaterais e 18% unilaterais <13cm. Entre as características histológicas, êmbolos carcinomatosos e a ausência de gradiente morfológico foram mais observados nos metastáticos. Na análise bivariada dos marcadores apenas CK7, CK20 e CDX2 mostraram diferenças significantes entre os grupos, entretanto houve muita sobreposição de resultados. Após análise multivariada foram selecionados: gradiente histológico e CK7 para formação de um novo algoritmo que definiu, com acurácia de 91%, que um tumor é metastático quando apresenta qualquer um dos aspectos: bilateral; unilateral e <13 cm; ausência de gradiente histológico; ou gradiente histológico presente com falta de expressão do CK7. Conclusão: tanto o algoritmo, como as reações imunoistoquímicas e os aspectos morfológicos são úteis no diagnóstico diferencial entre primário e metastático, porém não há nenhum dado discriminatório e, em alguns casos, somente a análise com equipe multidisciplinar pode definir o primário, reconhecendo as peculiaridades deste diagnóstico desafiador / Abstract: Primary ovarian mucinous carcinomas are uncommon and the most important differential is metastatic adenocarcinoma, mainly from gastrointestinal origin. Besides immunohistochemical profile, an algorithm determines, with a high accuracy, that unilateral and >13cm tumors are primary carcinomas and all the others, metastasis. Objective: to describe clinical and histopathological aspects of mucinous carcinomas, assessing the algorithm accuracy and immunohistochemical markers contributory to diagnosis. Methods: 76 mucinous carcinomas from our files (1994-2009) were revised; immunohistochemical reactions for CK7, CK20, Ca125, hormonal receptors (ER, PR), WT1, SMAD4, ?-catenin, CDX2 were performed by TMA. Results: 35 were ovarian primary tumors (group 1), 33 were metastasis (group 2). In eight cases the primary was not identified and these were excluded from statistic analysis. Most of the metastasis were from colorectal cancer (54%). Mean survival differed between the groups (65X35 months; p<0.0001). Agreement with the algorithm was 82.1%. In group 1, 85% were unilateral >13cm; in group 2, 61% were bilateral and 18% unilateral tumors <13cm. Different from group 1, common features in group 2 were vascular invasion and tumors without histological gradient. Bivariate analysis pointed out CK7, CK20 and CDX2 as main markers to distinguish both groups, but overlapping of the results was observed. After multivariate analysis, 2 aspects were selected: histological gradient and CK7; a new algorithm was designed and established with an accuracy of 91%, that a mucinous carcinoma is metastatic to the ovary when it shows one of the aspects: bilateral, or unilateral and <13cm, or without histological gradient, or presence of histological gradient but CK7 is negative. Conclusion: Algorithm and immunohistochemistry are useful, but there is no gold-standard marker. In some cases, only multidisciplinary evaluation can achieve reliable anatomo-clinical diagnosis, in this challenging situation / Doutorado / Anatomia Patologica / Doutora em Ciências Médicas

Orientadores: Cristina Laguna Benetti Pinto, Rose Luce Gomes do Amaral / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-28T09:13:50Z (GMT). No. of bitstreams: 1 GiraldoSouza_HelenaPatriciaDonovan_M.pdf: 2305470 bytes, checksum: add3640d7732bc5676b5b9eb414466b0 (MD5) Previous issue date: 2015 / Resumo: Introdução: Insuficiência Ovariana Prematura 46XX (IOP) é um estado de hipogonadismo, caracterizado por oligoamenorréia, infertilidade e deficiência estrogênica em mulheres abaixo de 40 anos. Mulheres com IOP deveriam ser tratadas com reposição estrogênica até pelo menos a idade da menopausa, para reduzir os sinais e sintomas do hipoestrogenismo e se possível, preservar a massa mineral óssea. Objetivos: Avaliar se Terapia Hormonal (TH) com baixa dose estrogênica é adequada para reduzir a perda de massa óssea de mulheres com IOP. Métodos: Estudo de corte transversal. Foram avaliadas 239 mulheres com IOP: 132 usando TH baixa dose (17-Beta-Estradiol 1mg + Noretisterona ou estrogênio conjugado 0,625mg + acetado de Medroxiprogesterona) e 107 mulheres sem TH. Todas responderam anamnese detalhada (idade, idade na última menstruação e idade no início de tratamento) e foram submetidas a avaliação de densidade mineral óssea (DMO) na coluna lombar (CL), colo femoral (CF) e fêmur total (FT) através da técnica DEXA. Resultados: As médias de idade, idade na última menstruação e de IMC para o grupo sem tratamento e para o grupo com TH foram 38,1 ± 6,1 e 36,8 ± 7,3 anos; 31,4 ± 7,3 e 30,7 ± 7,2 anos; 26,6 ± 7,1 e 25,8 ± 4,6, respectivamente (p=NS). A DMO média na CL foi 1,06 ± 0,15 e 1,00 ± 0,17g/cm2 (p=0,003), para CF 0,92 ± 0,15 e 0,89 ± 0,14 (p=0,0479) e FT de 0,92 ± 0,19 e 0,91 ± 0,13 g/cm2 (p=0,039), respectivamente para os grupos. Verificou-se DMO alterada na CL em 45,1% (35,2% Osteopenia e 9,8% Osteoporose) das mulheres sem tratamento e 60,1% (38,1 Osteopenia e 22% Osteoporose) quando usavam TH baixa dose (p=0,01). No CF 25,4% (21,5% Osteopenia e 3,9% Osteoporose) das mulheres sem tratamento e 29,6% (22,8% Osteopenia e 6,7% Osteoporose) quando usavam TH baixa dose (p=0,38) mostravam alteração e, para FT, em 32,35% (19,6% Osteopenia e 12,7% Osteoporose) das sem tratamento e 36,4% (21,2% Osteopenia e 15,2% Osteoporose) para TH de baixa dose (p=0,34). Conclusão: A TH de baixa dose não parece ser adequada para reduzir a perda de massa óssea de coluna lombar, colo de fêmur e fêmur total em mulheres com IOP / Abstract: Introduction: Premature Ovarian Insufficiency (POI) is a hypogonadism state, characterized by oligoamenorhea, infertility and estrogen deficiency in women below the age of 40. Women with POI should be treated with estrogen replacement until at least the age of menopause, in order to reduce the signs and symptoms of hypoestrogenism and if possible, preserve bone mineral density (BMD). Aim: To assess whether hormone therapy (HT) with low estrogen dose is sufficient to avoid bone mass loss in women with POI. Methods: Cross-sectional study. Two hundred and thirty nine women were evaluated: 132 using low dose TH (1 mg of 17-Beta-Oestradiol + Norethisterone or 0.625 mg of conjugated estrogen + medroxyprogesterone acetate) and 107 women without HT. Detailed history was obtained from the studied women (age, age of last menstrual period and age of onset of treatment) and were subjected to evaluation of bone mineral density (BMD) in the lumbar spine (LS), femoral neck (FN) and total femur (TF) through DEXA technique. Results: The mean age, age at last menstrual period and BMI for the untreated group and the group with HT were 38.1 ± 6.1 and 36.8 ± 7.3 years; 31.4 ± 7.3 and 30.7 ± 7.2 years; 26.6 ± 7.1 and 25.8 ± 4.6 respectively (p = NS). The mean LS BMD was 1.06 ± 0.15 and 1.00 ± 0,17g / cm2 (p = 0.003), CF 0.92 ± 0.15 and 0.89 ± 0.14 (p = 0.0479) and FT 0.92 ± 0.19 and 0.91 ± 0.13 g / cm2 (p = 0.039) respectively for the groups. BMD at LS was compromised in 45.1% (35.2% osteopenia and osteoporosis 9.8%) for women without treatment and 60.1% (38.1% osteopenia and 22% osteoporosis) low dose HT (p = 0.01). For the FN 25.4% (21.5% Osteopenia and Osteoporosis 3.9%) of women without treatment and 29.6% (22.8% Osteopenia and Osteoporosis 6.7%) for the ones in use of TH low dose, were compromised (p = 0 38). For TF compromise was found in 32.35% (19.6% osteopenia and 12.7% osteoporosis) of the untreated women and 36.4% (21.2% and 15.2% osteoporosis osteopenia) for low dose HT (p = 0.34). Conclusion: The low dose HT seems to be inadequate to reduce bone loss in the lumbar spine, femoral neck and total femur in women with IOP / Mestrado / Fisiopatologia Ginecológica / Mestra em Ciências da Saúde

Ovarian cancer is a malignancy characterized by poor prognosis, high levels of distress, and impaired quality of life (QOL). Investigation into the contributors to QOL is of psychological and prognostic significance in cancer. Contemporary stress theories and empirical accounts identify early life adversity and recent life stress as those sources which exert significant impact on physical and psychological health. To date, life stress research in cancer has yielded few designs which operationalize both indices of early life and recent life stress exposures. Moreover, despite the high-resolution stress data provided by the Life Events and Difficulties Schedule (LEDS) system, no studies to date comprehensively operationalize the early life adversity data obtained during each interview. Therefore, the proposed study is the first of its kind to comprehensively obtain ratings and examine effects of early life adversity data collected as part of the LEDS interview. It is also the first to examine independent influences of differentially timed life stress indices on psychological variables important to psychosocial functioning in ovarian cancer. Early life adversity was experienced by 43.1% of the sample. Adversity varied in content, number of occurrences, and severity. Ongoing difficulties, but not recent life events or early life adversity, were significantly associated with pre-surgical depression and QOL. Ongoing difficulties were also associated with lower depression, sleep, and QOL scores at all time-points. Early life adversity was associated with a poorer trajectory of sleep and QOL over the first year post-diagnosis. Findings are discussed with attention to behavioral and biological mechanisms. Applications to generative and cumulative theories of life stress are proposed. These findings lend support to the potential benefit of interventions aimed toward practical support and stress management in patients with ovarian cancer, as well as provide guidelines for use of early life adversity data obtained through the LEDS interview.

L’amélioration de la prise en charge personnalisée des cancers de l’ovaire nécessite le développement de nouvelles approches thérapeutiques et d’outils capables de prédire la réponse au traitement et la récidive. L’étude des miARNs cellulaires et circulants représente un champ d’investigation prometteur en oncologie, et ils pourraient rapidement constituer de nouveaux outils diagnostiques, pronostiques, voire thérapeutiques. Suite à la réalisation d’un criblage fonctionnel haut débit d’une banque de miARNs sur des lignées cancéreuses ovariennes, nous avons identifié plusieurs miARNs d’intérêt. Alors que l’utilisation des miARNs en tant qu’agents thérapeutiques n’est pas aujourd’hui envisageable, la caractérisation des cibles «pas-à-pas» de l’un de ces candidats, le miR-3622b-5p, en lien avec son action anti-tumorale au sens large (pro-apoptique, sensibilisation au cisplatine et anti-invasive) sur un panel de lignées cancéreuses ovariennes, a permis de renforcer l’intérêt de l’approche que nous développons qui consiste à reproduire l’effet de miARNs en ciblant les déterminants de leur action à l’aide de molécules inhibitrices, potentiellement utilisables en clinique. En parallèle, afin d’identifier globalement les cibles (leur aspect direct ou indirect vis-à-vis d’un miARN devenant alors secondaire) et réseaux associés des miARNs les plus prometteurs, identifiés lors du du screening, et en se basant initialement sur le premier apoptomiR identifié au laboratoire, le miR-491-5p, pour en faire la preuve de concept, une évaluation de l’approche méthodologique combinant des données d’approches multi-omiques a permis de conclure quant à l’intérêt de coupler les analyses trancriptomique et protéomique pour identifier de nouvelles cibles/voies en lien avec son action pro-apoptotique et ainsi proposer des associations pharmacologiques pertinentes et innovantes pour les cancers de l’ovaire. Par ailleurs, nous avons identifié une signature de miARN sérique capable d’identifier les patients susceptibles de présenter une résistance aux sels de platine et potentiellement aux inhibiteurs de PARP, préalablement à toute chimiothérapie et au moment de la récidive avant la seconde ligne de traitement. En résumé, ces résulats offrent de grandes promesses et placent les miARNs au coeur la médecine de précision de demain dans les cancers de l’ovaire. / Improving the personalized management of ovarian cancer requires the development of new therapeutic approaches and tools able to predict treatment response and recurrence. The study of cellular and circulating miRNAs represents a promising field of investigation in oncology, and they could quickly constitute new diagnostic, prognostic and even therapeutic tools. Following a high-throughput functional screening of a miRNA library on ovarian cancer lines, we identified several miRNAs of interest. While the use of miRNAs as therapeutic agents is not currently possible, the characterization of the targets «step-by-step»of one of these candidates, the miR-3622b-5p, with its broad anti-tumoral activity (pro-apoptic, sensibilisation to cisplatin and anti-invasive) on a panel of ovarian cancer lines, made it possible to reinforce the interest of the approach which we develop which consists in reproducing the effect of miRNAs by targeting the determinants of their action using inhibitory molecules, potentially usable in the clinic. In parallel, in order to globally identify the targets (their direct and indirect aspect next to a miRNA then becoming secondary) and associated networks of the most promising miRNAs, identified during the screening, and initially based on the first laboratory-identified apoptomiR, the miR-491-5p, to provide a proof of concept, an evaluation of the methodological approach combining data from multi-omic strategies led to the conclusion that it is useful to combine the trancriptomic and proteomic analyses to identify new targets/pathways related to its pro-apoptotic action and thus propose relevant and innovative pharmacological associations for ovarian cancers. In addition, we have identified a serum miRNA signature capable of identifying patients who may be resistant to platinum-based chemotherapy and potentially PARP inhibitors, prior to any chemotherapy and at the time of recurrence before the second line of treatment. In summary, these results offer great promise and place miRNAs at the heart of tomorrow's precision medicine in ovarian cancer.

Thesis (M. Sc. (Animal Production)) -- University of Limpopo / The objectives of this study were: 1) To determine the effect of oocyte retrieval techniques (slicing and aspiration) on the quality and quantity of cattle oocytes, 2) To evaluate the effect of different concentrations of hormones on the maturational rate of cattle oocytes selected by brilliant cresyl blue staining, 3) To evaluate fertilization rate and cleavage/embryonic development of oocytes with or without cumulus cells, and 4) To compare the effect of fresh and frozen thawed semen on the fertilization rate of cattle oocytes. In Experiment 1: oocytes were recovered from abattoir derived ovaries using slicing and aspiration. The recovered oocytes were exposed for 90 minutes to 26μM of brilliant cresyl blue (BCB) stain and classified according to the colour of their cytoplasm: BCB+ (oocytes with blue cytoplasm) and BCB- (unstained oocytes). There was no difference (P>0.05) in the quality of oocytes recovered using slicing (60.7 %) or aspiration (53.7 %) techniques. In experiment 2: The BCB selected and the non-selected immature oocytes were randomly allocated into medium 199 + 10 % fetal bovine serum (FBS) maturation media. The media was supplemented with three different concentrations of hormones as treatments (T). The T1 (0.5 μg/ml of follicle stimulating hormone (FSH), 5mg/ml of luteinizing hormone (LH) and 2 μg/ml of estradiol (E2) as the control group. Then, T2 (1 μg/ml of FSH, 6 mg/ml of LH and 2.5 μg/ml of E2) and T3 (1.5 μg/ml of FSH, 7 mg/ml of LH and 4.5 μg/ml of E2). Maturation rate of oocytes was determined by the protrusion of the first polar bodies 24 hours following in vitro maturation. Treatment 2 yielded higher (P<0.05) maturation rate for both BCB+ (65.6 %) and without BCB (60.3 %) oocytes with T1 giving lower (P<0.05) maturation rate for BCB+ (22 %) and without BCB (16 %) oocytes. However, BCB- oocytes had lower (P<0.05) polar body extrusion (3.03 %, 8.1 % and 2.2 %) for T1, T2 and T3, respectively. In Experiment 3: one group of the presumptive zygotes was denuded of cumulus cells and the other group was cultured with cumulus cells. The presumptive zygotes were in vitro cultured in SOF-BSA and changed to SOF-FBS after 48 hours. High fertilization/cleavage rate was observed in oocytes cultured with cumulus cells (29.0 %) compared to the denuded oocytes (20.0 %) for 2-4 cells stage. Day 7 blastocysts were more (P<0.05) on oocytes cultured with cumulus cells (32 %) compared to denuded oocytes (13 %). In experiment 4: The matured oocytes were fertilized using fresh and frozen thawed semen. The oocytes fertilized with frozen thawed semen obtained a better number of 2-4 cell cleavage (23 %) when compared to fresh semen (19 %). Oocytes that were fertilized with frozen thawed semen also obtained higher morula (13 %) and blastocyst (8 %) compared to fresh semen with morula (3.4 %) and blastocyst (2 %). In conclusion, immature oocytes that were exposed to BCB+ and cultured in M199 supplemented with 10 % FBS, 0.5 μg/ml of FSH, 5 mg/ml of LH and 2 μg/ml of E2 had a higher (P<0.05) number of matured oocytes (extrusion of first polar body) compared to those that were not exposed to BCB (no BCB). Oocytes that were cultured with cumulus cells yielded a higher (P<0.05) number of cleaved embryos compared to the denuded oocytes. Slicing yielded a higher (P<0.05) number of oocytes, however the quality of oocytes recovered was similar compared to those recovered by the aspiration technique (P>0.05). Oocytes fertilized with frozen thawed semen yielded higher (P<0.05) number of 2-4 cell, morula and blastocyst when compared with oocytes that were fertilized using fresh semen. Keywords: ovaries, oocytes, slicing, aspiration, COCs, BCB, polar body and cattle

Thesis (M. Sc. (Animal Production)) -- University of Limpopo, 2015 / The objectives of this study were: 1) To determine the effect of oocyte retrieval techniques (slicing and aspiration) on the quality and quantity of cattle oocytes, 2) To evaluate the effect of different concentrations of hormones on the maturational rate of cattle oocytes selected by brilliant cresyl blue staining, 3) To evaluate fertilization rate and cleavage/embryonic development of oocytes with or without cumulus cells, and 4) To compare the effect of fresh and frozen thawed semen on the fertilization rate of cattle oocytes. In Experiment 1: oocytes were recovered from abattoir derived ovaries using slicing and aspiration. The recovered oocytes were exposed for 90 minutes to 26μM of brilliant cresyl blue (BCB) stain and classified according to the colour of their cytoplasm: BCB+ (oocytes with blue cytoplasm) and BCB- (unstained oocytes). There was no difference (P>0.05) in the quality of oocytes recovered using slicing (60.7 %) or aspiration (53.7 %) techniques. In experiment 2: The BCB selected and the non-selected immature oocytes were randomly allocated into medium 199 + 10 % fetal bovine serum (FBS) maturation media. The media was supplemented with three different concentrations of hormones as treatments (T). The T1 (0.5 μg/ml of follicle stimulating hormone (FSH), 5mg/ml of luteinizing hormone (LH) and 2 μg/ml of estradiol (E2) as the control group. Then, T2 (1 μg/ml of FSH, 6 mg/ml of LH and 2.5 μg/ml of E2) and T3 (1.5 μg/ml of FSH, 7 mg/ml of LH and 4.5 μg/ml of E2). Maturation rate of oocytes was determined by the protrusion of the first polar bodies 24 hours following in vitro maturation. Treatment 2 yielded higher (P<0.05) maturation rate for both BCB+ (65.6 %) and without BCB (60.3 %) oocytes with T1 giving lower (P<0.05) maturation rate for BCB+ (22 %) and without BCB (16 %) oocytes. However, BCB- oocytes had lower (P<0.05) polar body extrusion (3.03 %, 8.1 % and 2.2 %) for T1, T2 and T3, respectively. In Experiment 3: one group of the presumptive zygotes was denuded of cumulus cells and the other group was cultured with cumulus cells. The presumptive zygotes were in vitro cultured in SOF-BSA and changed to SOF-FBS after 48 hours. High fertilization/cleavage rate was observed in oocytes cultured with cumulus cells (29.0 %) compared to the denuded oocytes (20.0 %) for 2-4 cells stage. Day 7 blastocysts were more (P<0.05) on oocytes cultured with cumulus cells (32 %) compared to denuded oocytes (13 %). In experiment 4: The matured oocytes were fertilized using fresh and frozen thawed semen. The oocytes fertilized with frozen thawed semen obtained a better number of 2-4 cell cleavage (23 %) when compared to fresh semen (19 %). Oocytes that were fertilized with frozen thawed semen also obtained higher morula (13 %) and blastocyst (8 %) compared to fresh semen with morula (3.4 %) and blastocyst (2 %). In conclusion, immature oocytes that were exposed to BCB+ and cultured in M199 supplemented with 10 % FBS, 0.5 μg/ml of FSH, 5 mg/ml of LH and 2 μg/ml of E2 had a higher (P<0.05) number of matured oocytes (extrusion of first polar body) compared to those that were not exposed to BCB (no BCB). Oocytes that were cultured with cumulus cells yielded a higher (P<0.05) number of cleaved embryos compared to the denuded oocytes. Slicing yielded a higher (P<0.05) number of oocytes, however the quality of oocytes recovered was similar compared to those recovered by the aspiration technique (P>0.05). Oocytes fertilized with frozen thawed semen yielded higher (P<0.05) number of 2-4 cell, morula and blastocyst when compared with oocytes that were fertilized using fresh semen. Keywords: ovaries, oocytes, slicing, aspiration, COCs, BCB, polar body and cattle

Case Report: Krukenberg tumor is a metastatic adenocarcinoma of the ovary that classically arises from the gastrointestinal tract, most often as a metastasis from the stomach as the primary origin, followed by colon. Krukenberg tumors are very rare malignant tumors of the ovary, only accounting for 1-2% of all ovarian malignancies. They tend to present with bilateral involvement. The most common presenting symptoms are abdominal pain, distention, and ascites, secondary to the large ovarian masses. Postmenopausal vaginal bleeding is a rare presenting symptom of a Krukenberg tumor. The diagnosis is commonly delayed until late in the disease progression. We present a case of a 77-year-old woman with stage IV metastatic adenocarcinoma of lower GI with mesenteric involvement and pulmonary nodules. Her disease was confirmed by mesenteric mass biopsy and was histologically CK20 positive, CDX positive, and CK7 negative. She underwent eighteen rounds of palliative chemotherapy with oral capecitabine (Xeloda) over the course of fifteen months. Sixteen months after the initial diagnosis, imaging uncovered a new cystic pelvic mass measuring 15x13x12 cm, decreased mesenteric mass, increasing liver lesion, metastasis to the left adrenal gland, and minimal ascites. She has had vaginal bleeding. Patient underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy and small bowel resection by gynecological oncologist. The left ovary was involved by metastatic adenocarcinoma, 15 cm, consistent with small bowel origin. The small bowel resection showed adenocarcinoma, 3.3 cm in size with serosal invasion arising in an adenoma. Patient is planned for chemotherapy with irinotecan in palliation. Our case demonstrates a rare case of small bowel adenocarcinoma later presenting as a Krukenburg tumor.

INTRODUCTION: Polycystic Ovary Syndrome (PCOS) is the most common endocrinopathy causing infertility in women of reproductive age. According to the Rotterdam criteria, a PCOS diagnosis should be given if at least two of the following are met: 1) hyperandrogenism; 2) oligo-anovulation; and 3) polycystic ovarian morphology. Previous studies analyzing the prevalence of PCOS have done so in unselected and clinical populations but few studies have attempted to characterize the syndrome and its long-term outcomes within Electronic Health Records using International Classification of Disease (ICD) codes. OBJECTIVES: With a hospital-based electronic health record dataset, this thesis seeks to: (1) characterize PCOS in reproductively aged women (18-34) using the diagnostic codes (ICD-9 and ICD-10) versus the Rotterdam criteria, (2) determine the prevalence of metabolic syndrome (MetS), Type 2 Diabetes, and cardiac events in women above age 35, (3) determine age of diagnosis for MetS and time to diagnosis of MetS. METHODS: The following 3 cohorts were queried on the Research Patient Data Registry (RPDR): 1) patients aged 18-34 with classic PCOS (phenotype A and B) but without an ICD diagnosis for PCOS, 2) patients aged 18-34 with a PCOS ICD-9/10 diagnosis and 3) patients above age 35 with a history or current diagnosis of PCOS. Their electronic health records (between January 1 , 2003 and December 31 , 2020) were ascertained from 9 Mass General Brigham institutions after IRB approval and analyzed on Software for Statistics and Data Science (STATA). RESULTS: Overall, RPDR identified 12,669 patients aged 18-34 who fit the Rotterdam criteria (under multiple phenotypes), 4646 of which had classic PCOS but lacked an ICD- 9/10 code for PCOS. RPDR also identified 9341 women aged 35 and above with a past or current diagnosis of PCOS. Hispanics/Latinas (18-34) were two times more likely to be undiagnosed when compared to Non-Hispanic Whites (OR: 2.25, 95% CI: 1.98-2.56). The prevalence of MetS, specified by a diagnostic code (277.7 or E88.81), and other cardiac conditions in women above age 35 were considerably lower than those found in the current literature. CONCLUSION: Databases such as RPDR allow for a detailed analysis of patient demographics, labs, procedures and diagnoses. Additionally, it allows for larger cohorts of patients matching more specific criteria to be ascertained. Future studies should compare the prevalence of individual features of MetS by ICD codes and analyze the cardiology reports to determine if the events are being reported but not codified. / 2023-11-30

Ces dernières années le rôle du stroma tumoral (microenvironnement) dans la progression tumorale. De même le rôle des cellules endothéliales et de la néo-angiogenèse a été illustré dans de multiples études conduisant à la mise en place des thérapeutiques anti-angiogéniques. Cependant il est possible qu’au delà de leur simple rôle comme transporteur d’oxygène et de nutriments les cellules endothéliales jouent un véritable rôle dans la biologie tumorale en sécrétant des substances bioactives (cytokines, microparticules…). Ces médiateurs sont les acteurs actifs d’un dialogue entre cellules tumorales et cellules du stroma. Dans ce travail de thèse nous nous sommes intéressés au rôle particulier des microparticules. Nous avons pu montrer que les microparticules des cellules endothéliales avaient un effet pro-tumoral sur les cellules des cancers de l’ovaire et du sein. Elles étaient capable d’induire une tradition epithélio-mésenchymateuse. Dans la seconde étude nous avons montré que les cellules tumorales sécrétaient des microparticules capable d’activer la voie de signalisation ont/beta-catenin dans les cellules endothéliales par le recrutement de Rac1 et PAK. / In our study, we showed that microparticles participate to a complex dialogue between cancer and ECs. Our main finding showed the ability of MPs mediated cross-talk between cancer and ECs to functionalize an activated angiocrine pro-tumoral endothelial niche. Using endothelial Akt activation as a readout, we were able to differentiate MPs from cells with mesenchymal from cells with epithelial traits. Our data showed that MPs from mesenchymal-like cell lines were able to promote an activation of ECs through Akt phosphorylation, compared to MPs from epithelial-like cell lines. The overexpression of Arf6 in activated ECs is associated with quantitative changes of EC-MPs. Additionally, we were interested in determining the mechanisms that derive the activation of ECs toward supporting tumor growth and expansion. Here we showed that ovarian cancer MPs trigger β-catenin activation in ECs by inducing the upregulation of Wnt/bcatenin target genes and increasing the angiogenic proprieties. Interestingly, the activation of bcatenin in ECs was Wnt/Frizzled independent; but dependent on VE-cadherin localization disruption, bcatenin integrin activation and MMP activity.

Cette thèse de doctorat est une étude de la biologie et de la dynamique des microARN circulants, démontrant leur potentiel comme biomarqueurs pour l’amélioration de la surveillance et de l’évaluation pronostique dans le cancer. Il s’agit des ARN non codants simple-brin, d'environ 19-25 nt de longueur. Ils jouent un rôle clé dans la régulation de l'expression génomique au niveau post-transcriptionnel, en ciblant et réprimant la traduction des ARNm par complémentarité partielle avec leur 3'-UTR. Ils sont également libérés dans le milieu extracellulaire, la circulation et les liquides biologiques. Leur stabilité remarquable et leur diversité dans la circulation, ainsi que leur provenance maligne ou normale, en font des candidats biomarqueurs intéressants, reflétant potentiellement l'état et la dynamique d’une tumeur. Nous nous sommes focalisés sur les carcinomes ovariens (OvCa) et nasopharyngés (NPC), essayant d'élucider la relation entre les miARN plasmatiques et le pronostique des OvCa après une première ligne de traitement, ainsi que d’évaluer leur utilité dans la détection d’une réponse précoce des NPC au traitement. Le carcinome ovarien séreux est la malignité gynécologique la plus agressive. L'absence de symptômes précoces et l'insuffisance des moyens modernes pour détecter la maladie résiduelle et évaluer les résultats du traitement signalent le besoin de nouveaux biomarqueurs diagnostiques et pronostiques. En utilisant des prélèvements plasmatiques séquentiels OvCa avant et après traitement, nous avons étudié un groupe de miARN, en comparaison à des lésions pelviennes bénignes et des femmes en bonne santé. MiR-200b avait une concentration nettement plus élevée dans les échantillons malins avant traitement par rapport aux deux groupes non cancéreux. L'analyse pré- et post-traitement de miR-200b en parallèle avec le biomarqueur standard CA125 a révélé des variations distinctes et une corrélation significative de la variation de miR-200b avec le temps de rémission (PFS). Nous concluons que miR-200b pourrait éventuellement être utilisé comme biomarqueur supplémentaire pour l'estimation de la rémission à la fin du traitement. Le carcinome nasopharyngé (NPC) d'autre part est une tumeur constamment associée à une infection latente des cellules malignes par le virus d’Epstein-Barr (EBV), présentant une distribution géographique particulière. La position de la tumeur rend difficile l'approche chirurgicale, les biomarqueurs évaluant différentes approches thérapeutiques étant de grande importance. Etudiant une nouvelle forme orale de l'agent démethylant 5-azacytidine, démontrée prometteuse pour un tiers des patients NPC qui l’ont reçue, nous avons essayé d'évaluer son impact sur l'expression des miARN et des protéines virales. Malgré l'infection virale latente, les miARN viraux sont abondamment exprimés. En traitant pendant deux semaines quatre modèles de tumeur NPC développés in vivo, nous avons observé une réponse nette dose-dépendante dans les deux. L'analyse protéique a montré une induction de la protéine activatrice du cycle lytique BZLF1, renforçant les preuves antérieures d'induction partielle du cycle lytique viral par la 5-azacytidine. L'analyse des miARN a confirmé l'expression robuste des miARN BART et l'absence des miARN BHRF1 dans les NPC sans traitement. Lors du traitement, nous avons détecté les miR-BHRF1 à la fois dans la tumeur et le plasma des souris traitées. Cette induction a été validée par un traitement ultérieur d'une semaine qui a aussi mis en évidence l'induction de l’expression de l'ARNm de BHRF1, transcrit par le locus situé parmi les miARN BHRF1. Une induction de ces miARN a été observée après traitement par des agents de chimiothérapie, suggérant une utilité clinique potentielle des miR-BHRF1 comme biomarqueurs pour l’évaluation précoce de l’efficacité du traitement. Notre objectif actuel est de valider cette induction par chimiothérapie et étendre nos études au plasma humain. / This doctorate thesis provides an insight into the biology and dynamics of circulating microRNAs, demonstrating their potential to become crucial biomarkers for a better surveillance and prognosis of cancer. MicroRNAs (miRNAs) are small, single-stranded non-coding RNAs, 19-25 nt long, with a key role in the post-transcriptional regulation of gene expression, repressing the translation of target mRNAs through partial base-pair complementarity with their 3’-UTR. They can be released in the extracellular medium, being protected from RNases by association with various transporters and reach body fluids and circulation, participating in intercellular communication. Their remarkable stability and manageable diversity in circulation, as well as the fact that they derive from both malignant and normal cells make them very attractive biomarker candidates, potentially reflecting tumor state and dynamics. We have been focusing on ovarian (OvCa) and nasopharyngeal (NPC) carcinomas, attempting to elucidate the relation between plasma miRNAs and the prognosis of OvCa after first-line treatment, as well as to evaluate their use in the detection of early response of NPC tumors to treatment. Serous epithelial ovarian carcinoma is the most frequent ovarian and the most aggressive gynecologic malignancy. The absence of early symptoms and the insufficiency of modern means to accurately map residual disease and assess treatment outcome highlight the need for new diagnostic and prognostic biomarkers. Using sequential plasma samples from OvCa patients before and after first-line treatment, we studied a pre-selection of miRNAs, comparing them to samples from benign pelvic lesions and healthy women. MiR-200b exhibited a distinct higher concentration in malignant samples before treatment compared to both non-cancerous groups. Pre- and post-treatment assessment of miR-200b in parallel with the standard biomarker CA125 revealed distinct variations and a significant correlation of miR-200b variation with the progression-free survival (PFS) of the patient. We suggest that miR-200b could eventually be used as a supplementary biomarker for estimation of the remission upon treatment completion. Nasopharyngeal carcinoma (NPC) on the other hand is a tumor consistently associated to latent Epstein-Barr virus (EBV) infection of the malignant cells, presenting a unique geographical incidence pattern. The deep position of the tumor makes it tough to access surgically, with biomarkers assessing different therapeutic approaches being greatly needed. Studying a new oral form of the demethylating agent 5-azacytidine, proven to be promising for one third of NPC patients receiving it as a monodrug, we attempted to identify impact of the drug on the expression of viral miRNAs and proteins. Despite the latent viral infection, viral miRNAs are abundantly expressed, attracting interest in EBV-associated malignancies. Treating four in vivo developed NPC tumor models for two weeks, we observed clear response in two of them, in a dose-dependent manner. Protein analysis showed an induction of the immediate-early lytic protein BZLF1, solidifying previous evidence of partial activation of the viral lytic cycle by 5-azacytidine. MiRNA analysis confirmed robust expression of BART and absence of BHRF1 miRNAs at baseline status of NPC. Upon treatment, we observed an induction of BHRF1 miRNAs in both tumor and plasma of treated mice. This induction was successfully validated in a following one-week treatment and completed by a recorder de novo expression of the BHRF1 mRNA, transcribed within the BHRF1 miRNA loci. A weaker induction of BHRF1 miRNAs was also recorded after treatment with standard chemotherapeutic agents, suggesting a potential clinical utility of these miRNAs as circulating biomarkers for detection of early response to treatment. We are further working to confirm this induction by chemotherapy and extend our study to plasma samples derived from treated patients.