Spinale Meningeome machen ca. 30% von allen intradural-extramedullären Läsionen aus. Die Literatur, die sich mit spinalen Meningeomen beschäftigt hat, ist rar. Bisher ist nur wenig über das molekulare Profil von spinalen Meningeomen und seine klinischen Auswirkungen bekannt. Fragestellung: 1. Was sind die Prädilektionsstellen für spinale Meningeome und korrelieren diese mit dem postoperativen neurologischen Status? 2. Was sind die Risikofaktoren für die Entstehung eines Tumorrezidivs? 3. Welche Faktoren minimieren die Wahrscheinlichkeit einer vollständigen Resektion der spinalen Meningeome? 4. Lassen sich Meningeome anhand ihres molekularen Profils klassifizieren? Material und Methoden: Diese retrospektive Arbeit befasste sich mit Patienten, welche an einem spinalen Meningeom im Zeitraum von 1993 bis 2020 in der Klinik für Neurochirurgie am Universitätsklinikum „Carl Gustav Carus“ operiert wurden. Dabei wurde besonderes Augenmerk auf die klinischen und genomischen Merkmale der spinalen Meningeome gelegt. Insgesamt wurden 101 Patienten mit 104 spinalen Meningeomen in diese Untersuchung eingeschlossen. Deren Krankenakten systematisch erfasst und statistisch ausgewertet wurden. Klinisch, bildmorphologische sowie histopathologische Daten wurden analysiert. Des Weiteren wurden klinische Parameter mit Sequenzierungsergebnissen in einer gut charakterisierten Kohorte von 47 Patienten korreliert. Ergebnisse: Das mediane Alter aller Patienten in dieser Arbeit betrug 69 Jahre. Die Mehrheit der Tumore in dieser Studie war in der Brustwirbelsäule lokalisiert (56,7%). Die häufigsten neurologischen Symptome waren sensomotorische Defizite und Gangstörungen. Meningeome der BWS gingen am häufigsten mit präoperativen neurologischen Defiziten einher (p=0,005). Lediglich bei einem Patienten (0,9 %) wurde eine permanente, chirurgisch-bedingte, neurologische Verschlechterung beobachtet. Es ließ sich feststellen, dass Foramen magnum-Meningeome im Vergleich mit Meningeomen anderer Lokalisationen am häufigsten mit postoperativen neurologischen Defiziten assoziiert waren (p=0,01). Insgesamt konnte eine Verbesserung des funktionellen Outcomes durch die chirurgische Intervention gezeigt werden (67 von 104, p=0,00001). Damit profitieren die Patienten von einer Operation. Unabhängig vom präoperativen Status ist postoperativ mit einer Verbesserung des Outcomes zu rechnen. Es ließ sich in dieser Arbeit kein Einfluss von Resektionsgrad, histologischen Grad und Subtyp sowie Lokalisation des Tumors auf das Rezidiv nachweisen. Ein Ki-67-Index > 5% hat sich in dieser Untersuchung als einziger Prädiktor für ein Rezidiv spinaler Meningeome bewährt (p=0,0436). Die molekulare Untersuchung von 47 Patienten mit verfügbarem Material zeigt, dass AKT1-Mutationen ein häufiges genomisches Ereignis bei spinalen Meningeomen darstellen. Die Mehrzahl der AKT1-mutierten Meningeome trat bei männlichen Patienten auf (p=0,0175), war in der Halswirbelsäule, ventral des Rückenmarks lokalisiert (p=0,0304 bzw. p=0,0044) und wies eine meningotheliale Histologie auf (p=0,0339). Bezüglich der NF2-mutierten spinalen Meningeome konnte gezeigt werden, dass diese vor allem in der Brustwirbelsäule lokalisiert sind. Sie waren alle AKT1-Wildtyp-Meningeome und traten ausschließlich bei weiblichen Patienten auf. Ein bisher nicht beschriebenes Merkmal ist, dass alle verkalkten Meningeome in unserer Kohorte eine NF2-Mutation aber keine AKT1-Mutation aufwiesen (p=0,0061). Diese Ergebnisse zeigen abermals eine starke Korrelation zwischen den klinischen und genomischen Parametern spinaler Meningeome. Schlussfolgerung: Die Prognose der Patienten mit spinalen Meningeomen nach vollständiger Resektion ist sehr gut. Die Resektion spinaler Meningeome geht mit niedrigen Komplikationsraten sowie einem guten langfristigen funktionellen Ergebnis einher. Diese Studie verbessert unser Verständnis der Pathobiologie spinaler Meningeome und kann das Design klinischer Studien optimieren. Darüber hinaus kann die ungünstige Lokalisation und Konsistenz häufig die vollständige Resektion behindern, was bei einigen Autoren als ein unabhängiger Risikofaktor für ein Tumorrezidiv gilt. Angesichts dieser chirurgischen Herausforderungen könnte die Aufnahme betroffener Patienten in Studien, in denen die Wirksamkeit von AKT-Inhibitoren (z.B. Afuresertib, ClinicalTrials.gov NCT02523014) untersucht wird, sinnvoll sein. Ein solcher Ansatz erfordert das Screening spinaler Meningeomen auf AKT1-Mutationen. Zusammenfassend lässt sich sagen, dass ein Großteil spinaler Meningeome eine AKT1-Mutation und eine weitere, noch größere Gruppe, eine NF2-Mutation aufweisen und daher möglicherweise einer zielgerichteten Therapie zugänglich sind. Weitere Studien sind erforderlich, um den Einfluss relevanter Mutationen auf das Rezidivverhalten zu untersuchen und Meningeompatienten zu identifizieren, die den größtmöglichen Nutzen aus gezielten Therapiestrategien ziehen können.:INHALTSVERZEICHNIS III 1 EINLEITUNG 9 1.1 Definition 9 1.2 Epidemiologie 9 1.3 Lokalisation und Ursprung 9 1.4 Ätiologie 11 1.5 Klinisches Erscheinungsbild 11 1.6 Bildgebung 13 1.7 Klassifizierung und Subtypen 14 1.7.1 WHO-Grad 1 15 1.7.2 WHO-Grad 2 15 1.7.3 WHO-Grad 3 16 1.8 Meningeomwachstum und -rezidiv 16 1.9 Therapie spinaler Meningeome 17 1.9.1 Chirurgische Therapie 17 1.9.2 Radiotherapie 18 1.9.3 Chemotherapie 18 1.10 Verlaufskontrolle 18 1.11 Molekulare Alterationen in Meningeomen 19 1.11.1 NF2-Mutation 20 1.11.2 AKT1E17K-Mutation 20 2 ZIELSETZUNG UND BEDEUTUNG DER STUDIE 22 3 METHODIK 23 3.1 Studientyp 23 3.2 Patientenpopulation und Gewebeproben 23 3.3 Diagnostik und Klassifizierung 23 3.4 Prä- und postoperativer klinischer Zustand 24 3.5 Operation 24 3.6 Nachsorge 25 3.7 Neuropathologische Untersuchung 25 3.8 DNA-Sequenzierung und molekulare Charakterisierung 25 3.9 Statistische Analyse 27 4 ERGEBNISSE 28 4.1 Beschreibung der Patientenkohorte 28 4.2 Demographische Daten 28 4.2.1 Geschlechtsverteilung 28 4.2.2 Patientenzahl / Alter 29 4.2.3 Nebenerkrankungen 30 4.3 Bildgebung 32 4.4 Tumorlokalisation 33 4.4.1 Lokalisation nach Wirbelsäulenabschnitten 33 4.4.2 Lokalisation in Bezug auf das Myelon 34 4.4.3 Lokalisation in Bezug auf die Dura 34 4.5 Beispiele von spinalen Meningeomen 34 4.5.1 Fall 1 35 4.5.2 Fall 2 36 4.5.3 Fall 3 37 4.6 Chirurgische Therapie 37 4.6.1 Zugangsarten 38 4.6.2 Resektionsgrad 39 4.7 Histologie 39 4.8 Perioperative Komplikationen und Mortalität 40 4.9 Prä- und postoperativer neurologischer Zustand 41 4.9.1 Klinischer Zustand in Korrelation mit der Lokalisation des Meningeoms 43 4.10 Follow-up 49 4.10.1 Einfluss der Meningeomlokalisation auf das progressionsfreie Überleben (PFS) 51 4.10.2 Einfluss der Operationsradikalität und der Histologie auf die Rezidivrate 53 4.10.3 Einfluss des Proliferationsindex Ki-67 auf das Rezidiv 56 4.11 AKT1-Mutation 60 4.12 NF2-Mutation 62 5 DISKUSSION 65 5.1 Klinische Analyse 65 5.2 Molekulare Analyse 72 6 ZUSAMMENFASSUNG 76 7 SUMMARY 79 8 LITERATURVERZEICHNIS 81 9 DANKSAGUNG 91 / Spinal meningiomas account for approximately 30% of all intradural extramedullary lesions. The literature dealing with spinal meningiomas is scarce. Little is known about the molecular profile of spinal meningiomas and its clinical implications. This retrospective study describes the clinical and genomic characteristics of patients with spinal meningioma. Particular attention was given to the clinical and genomic characteristics of spinal meningiomas. Question: 1. What are the predilection sites for spinal meningiomas and how do they correlate with the postoperative neurological status? 2. What are the risk factors for tumor recurrence? 3. What factors minimize the likelihood of complete resection of spinal meningiomas? 4. Can NF2 and AKT1 mutant meningiomas be distinguished from each other on the basis of clinical or histological features? Material and Methods: A total of 101 patients with 104 spinal meningiomas, who underwent surgery in the period from 1993 to 2020 in the Department of Neurosurgery at the University Hospital 'Carl Gustav Carus' were included in this study. Their medical records were systematically reviewed and statistically evaluated. Clinical, image morphological and histopathological data were analyzed. Furthermore, clinical parameters were correlated with sequencing results in a well-characterized cohort of 47 patients. Results: The median age of all patients in this work was 69 years. The majority of tumors in this study were located in the thoracic spine (56.7%). The most common preoperative neurological symptoms were sensorimotor deficits and gait disturbances. Meningiomas of the thoracic spine were most frequently associated with preoperative neurological deficits (p=0.005). Consequently, permanent surgery-related neurological deterioration was rarely observed in this series (0.9% of cases), while surgical intervention significantly improved patients’ functional outcome (67 out of 104 patients p=0.00001). Regardless of the preoperative status, an outcome improvement can be expected postoperatively. In this work, neither the degree of resection, nor the histological degree or tumor localization influenced tumor recurrence, while a Ki-67 index of > 5% was the only predictor for recurrence in spinal meningiomas (p=0.0436). Furthermore, we correlated clinical parameters with the molecular status in a subset of 47 patients. Herein we show that AKT1 mutations are a common genomic event in spinal meningiomas (21.2%). The majority of AKT1-mutated meningiomas occurred in male patients (p=0.0175), were located in the cervical spine and anterior to the spinal cord (p=0.0304 and p=0.0044, respectively), and had meningothelial histology (p=0.0339). In contrast, NF2-mutated spinal meningioma were all located in the thoracic spine and occurred only in female patients. Moreover, all calcified meningiomas were NF2-mutant (p=0.0061). Our results show a significant correlation between the AKT1/NF2 mutation status with the histological subtype and the meningioma localization. Conclusion: The prognosis of a spinal meningioma patients treated with complete resection is very good, with low complication rates and a good long-term functional outcome. This study improves our understanding of the pathobiology of spinal meningiomas and may optimize designing clinical trials. In addition, the unfavorable location and consistency can often impede complete resection, which some authors consider to be an independent risk factor for tumor recurrence. Given these surgical challenges, enrolling affected patients in trials evaluating the efficacy of AKT inhibitors (e.g. afuresertib, ClinicalTrials.gov NCT02523014) of clinical interest. Such an approach requires the screening of spinal meningiomas for AKT1 and NF2 mutations. Taken together, we have identified two predominant molecular subgroups in WHO-grade 1 SM, characterized by AKT1 and NF2 mutations. Both mutations are mutually exclusive and are associated with distinct patient characteristics and tumor features. AKT1-mutant meningiomas originate in the cervical spine ventrally to the spinal cord, are almost exclusively associated with meningothelial histology and exhibit no calcifications on imaging. In contrast, NF2-mutant meningiomas show strong female gender predominance, arise with a wider anatomic distribution, although most frequently in the thoracic spine dorsally to the spinal cord, and can be calcified while displaying variable histologic subtypes. Further studies are needed to investigate the impact of relevant mutations on recurrence and to identify patients who can derive maximum benefit from targeted therapy strategies.:INHALTSVERZEICHNIS III 1 EINLEITUNG 9 1.1 Definition 9 1.2 Epidemiologie 9 1.3 Lokalisation und Ursprung 9 1.4 Ätiologie 11 1.5 Klinisches Erscheinungsbild 11 1.6 Bildgebung 13 1.7 Klassifizierung und Subtypen 14 1.7.1 WHO-Grad 1 15 1.7.2 WHO-Grad 2 15 1.7.3 WHO-Grad 3 16 1.8 Meningeomwachstum und -rezidiv 16 1.9 Therapie spinaler Meningeome 17 1.9.1 Chirurgische Therapie 17 1.9.2 Radiotherapie 18 1.9.3 Chemotherapie 18 1.10 Verlaufskontrolle 18 1.11 Molekulare Alterationen in Meningeomen 19 1.11.1 NF2-Mutation 20 1.11.2 AKT1E17K-Mutation 20 2 ZIELSETZUNG UND BEDEUTUNG DER STUDIE 22 3 METHODIK 23 3.1 Studientyp 23 3.2 Patientenpopulation und Gewebeproben 23 3.3 Diagnostik und Klassifizierung 23 3.4 Prä- und postoperativer klinischer Zustand 24 3.5 Operation 24 3.6 Nachsorge 25 3.7 Neuropathologische Untersuchung 25 3.8 DNA-Sequenzierung und molekulare Charakterisierung 25 3.9 Statistische Analyse 27 4 ERGEBNISSE 28 4.1 Beschreibung der Patientenkohorte 28 4.2 Demographische Daten 28 4.2.1 Geschlechtsverteilung 28 4.2.2 Patientenzahl / Alter 29 4.2.3 Nebenerkrankungen 30 4.3 Bildgebung 32 4.4 Tumorlokalisation 33 4.4.1 Lokalisation nach Wirbelsäulenabschnitten 33 4.4.2 Lokalisation in Bezug auf das Myelon 34 4.4.3 Lokalisation in Bezug auf die Dura 34 4.5 Beispiele von spinalen Meningeomen 34 4.5.1 Fall 1 35 4.5.2 Fall 2 36 4.5.3 Fall 3 37 4.6 Chirurgische Therapie 37 4.6.1 Zugangsarten 38 4.6.2 Resektionsgrad 39 4.7 Histologie 39 4.8 Perioperative Komplikationen und Mortalität 40 4.9 Prä- und postoperativer neurologischer Zustand 41 4.9.1 Klinischer Zustand in Korrelation mit der Lokalisation des Meningeoms 43 4.10 Follow-up 49 4.10.1 Einfluss der Meningeomlokalisation auf das progressionsfreie Überleben (PFS) 51 4.10.2 Einfluss der Operationsradikalität und der Histologie auf die Rezidivrate 53 4.10.3 Einfluss des Proliferationsindex Ki-67 auf das Rezidiv 56 4.11 AKT1-Mutation 60 4.12 NF2-Mutation 62 5 DISKUSSION 65 5.1 Klinische Analyse 65 5.2 Molekulare Analyse 72 6 ZUSAMMENFASSUNG 76 7 SUMMARY 79 8 LITERATURVERZEICHNIS 81 9 DANKSAGUNG 91

BACKGROUND:In breast cancer endocrine therapy, post-therapy Ki-67 assay of biopsy material predicts recurrence-free survival but is invasive and prone to sampling error. 18F]Fluorodeoxyglucose (FDG) positron emission tomography (PET) has shown an early agonist or 'flare' response to tamoxifen and estradiol, but has not been tested in response to estrogen-lowering aromatase inhibitors (AIs). We hypothesized that decreased agonistic response to AIs would result in early FDG uptake decline. We also measured early response to trastuzumab (T), another targeted agent for breast cancer with differing mechanisms of action. Our study was designed to test for an early decline in FDG uptake in response to AI or T and to examine association with Ki-67 measures of early response.METHODS:Patients with any stage of newly diagnosed or recurrent breast cancer were eligible and enrolled prior to initiation (or resumption) of AI or T therapy. FDG PET and tissue biopsy were planned before and after 2 weeks of AI or T therapy, with pretreatment archival tissue permitted. Cutoffs of greater than or equal to]20% decline in standardized uptake value (SUV) as FDG PET early response and less than or equal to]5% post-treatment expression as Ki-67 early response were defined prior to analysis.RESULTS:Forty-two patients enrolled, and 40 (28 AI, 12 T) completed serial FDG-PET imaging. Twenty-two patients (17 AI, 5 T) had newly diagnosed disease, and 23 (14 AI, 9 T) had metastatic disease (5 newly diagnosed). Post-treatment biopsy was performed in 25 patients (63%) and was either refused or not feasible in 15. Post-treatment biopsy yielded tumor in only 17/25 cases (14 AI, 3 T). Eleven of 14 AI patients with post-therapy tissue showed FDG PET early response, and there was 100% concordance of PET and post-therapy Ki-67 early response. For the T group, 6/12 showed an FDG PET early response, including 2/3 patients with post-therapy biopsy, all with Ki-67 >5%.CONCLUSIONS:Substantial changes in FDG PET SUV occurred over 2 weeks of AI therapy and were associated with low post-therapy proliferation. SUV decline was seen in response to T, but few tissue samples were available to test association with Ki-67. Our results support further investigation of FDG PET as a biomarker for early response to AI therapy.

Dans un premier temps, nous avons réalisé des revues systématiques de la littérature avec méta-analyses des données de survie. Ceci nous a conduits à sélectionner 4 marqueurs de mauvais pronostic pour la survie des CBNPC: le récepteur au facteur de croissance épidermique (EGF-R), un autre récepteur de cette famille (c-erbB-2) ainsi que deux autres facteurs potentiellement témoins de leur activité, Ki-67 (impliqué dans la prolifération) et le nombre des micro-vaisseaux (témoins de la néoangiogenèse). Dans une deuxième phase, nous avons étudié au laboratoire diverses questions sur des tumeurs bronchiques invasives. Premièrement, nous avons investigué le mécanisme de surexpression d’EGF-R et de c-erbB-2 et évalué si des anomalies génétiques pouvaient prédire cette surexpression, en recourant à des techniques d’immunohistochimie et de FISH. Ceci nous a permis d’observer que, si la majorité des CBNPC réséqués présentent des anomalies génétiques d’EGF-R et/ou de c-erbB-2, une amplification de ces gènes n’est présente que dans une minorité d’entre eux et n’est pas strictement corrélée à l’expression protéique. D’autre part, la survie de ces patients exprimant ou ayant une anomalie génique d’EGF-R et/ou c-erbB-2 est plus courte sans atteindre le seuil de signification statistique. Deuxièmement, nous avons recherché sur des tumeurs opérées d’éventuels liens entre les expressions d’EGF-R, de c-erbB-2 et de Ki-67. Aucune corrélation n’a été mise en évidence entre l’expression de ces 3 facteurs. Par contre, chez ces patients, l’expression de Ki-67 dans la tumeur s’est avérée être un facteur de mauvais pronostic pour la survie. Troisièmement, nous avons voulu savoir si un de ces marqueurs (EGF-R) présentait une valeur pronostique dans un groupe plus restreint de tumeurs plus avancées, les CBNPC de stade III. Pour mener cette recherche sur des biopsies, nous avons d’abord démontré que l’évaluation des marqueurs biologiques (EGF-R, c-erbB-2 et Ki-67) sur biopsie ne différait pas de celle réalisée sur des tumeurs réséquées. Comme les résultats étaient équivalents, nous avons pu étudier EGF-R sur les biopsies de CBNPC au stade III et montrer qu’EGF-R n’était pas un facteur pronostique pour la survie dans ce groupe assez homogène de tumeurs avancées. Dans la dernière phase, nous avons étudié des lésions représentatives des différents stades prénéoplasiques et néoplasiques précoces radiooccultes. Ces lésions ont été prélevées lors d’examens endoscopiques de photodétection. EGF-R, c-erbB-2, Ki-67 et le nombre des micro-vaisseaux ont été étudiés par immunohistochimie dans ces différents stades de lésions prénéoplasiques et néoplasiques précoces. Nous avons observé qu’EGF-R et Ki-67 sont statistiquement plus exprimés dans les dysplasies sévères et les carcinomes in que dans les dysplasies légères suggérant que, au moins pour ces 2 marqueurs, les dysplasies sévères se rapprochent plus des carcinomes in situ que des dysplasies légères. Alors que l’expression d’EGF-R est présente dès le stade de dysplasie sévère, une augmentation du nombre des micro-vaisseaux n’est présente qu’au stade de tumeurs micro-invasives. C-erbB-2 n’est quant à lui pas exprimé dans ces lésions bronchiques prénéoplasiques et néoplasiques précoces. En conclusion, les facteurs biologiques, EGF-R, c-erbB-2 et Ki-67 et le nombre des micro-vaisseaux s’avèrent des facteurs de mauvais pronostic dans le CBNPC. La surexpression d’EGF-R et de c-erbB-2 dans les cancers réséqués résulte très rarement d’une amplification génique et nous n’avons pas trouvé dans ces tumeurs de corrélation entre l’expression des marqueurs moléculaires étudiés. Dans les tumeurs plus avancées de stade III, EGF-R n’est pas un facteur discriminant pour le pronostic. Les anomalies de certains de ces marqueurs (EGF-R et Ki-67) apparaissent précocement, dès les stades prénéoplasiques, avec un seuil se situant entre les lésions bronchiques de bas et de haut grades. La néoangiogénèse, évaluée par le nombre des micro-vaisseaux, s’observe à partir des cancers micro-invasifs tandis que c-erbB-2 n’apparaît qu’au stade invasif. Dans la séquence d’apparition des anomalies génétiques conduisant au cancer invasif, l’atteinte d’EGF-R précède la néoangiogénèse.

<p>Breast cancer is one of the most common malignancies in women. Due to early detection and the use of screening programs approximately 60% of all new cases lack lymph node involvement. Today, a substantial proportion of these women will be offered adjuvant systemic chemotherapy. However, better proliferation markers are needed to predict patient outcome and to avoid overtreatment. </p><p>Cyclin A, cyclin E and Ki-67 are all markers for proliferation and involved in the regulation of the cell cycle. Overexpression has been associated with disease recurrence in several studies, but the results have not been consistent. However, none of these studies has investigated aberrant expression of cyclin E (the expression of cyclin E during phases of the cell cycle other than late G1 and early S-phase). Studies have shown that aberrant cyclin E might provide additional prognostic information compared to cyclin E alone.</p><p>The aims of this thesis were 1.to investigate the prognostic value of cyclin A, cyclin E and aberrant cyclin E in early breast cancer. 2.to validate the tissue microarray (TMA) technique for cyclin A and 3.to define the most optimal cut-off values for cyclin A and Ki-67.</p><p>We found that the agreement of TMA and large section results was good with kappa values 0.62-0.75 and that the reproducibility of the two readers’ results was good or even very good, with kappa values 0.71 – 0.87. </p><p>The optimal cut-off value for cyclin A average was 8% and for cyclin A maximum value 11%. The corresponding values for Ki-67 were 15 and 22%. </p><p>Neither cyclin E nor aberrant cyclin E was a prognostic factor in low-risk node negative breast cancer patients. </p><p>Finally, we conclude that cyclin A is a prognostic factor in node negative breast cancer (univariate analysis average value OR=2.9 95% CI 1.8-4.6; maximum value OR=3.7 95% CI 2.3-5.9).</p>

Es existiert bereits eine Vielzahl von tierexperimentellen Studien bezüglich Einflussfaktoren auf die adulte Neurogenese. Nachdem die Teilungsfähigkeit von neuralen Stammzellen Ende der 1990er Jahre auch im adulten humanen Gehirn nachgewiesen wurde, war es das Ziel der vorliegenden Arbeit, adulte Neurogenese bei psychischen Erkrankungen zu quantifizieren bzw. den Ein-fluss medikamentöser Therapien auf die adulte Neurogenese zu untersuchen. Diese Studie stellt dabei die bislang einzige Arbeit dar, die sich mit der humanen adulten Neurogenese bei psychischen Erkrankungen beschäftigt. Mittels Doppelfärbungen von Ki67 und BrdU an Mausgewebe wurde zunächst nachgewiesen, dass das Ki67-Antigen ein zuverlässiger Marker für sich teilende Zellen ist, woraufhin die Färbeprozedur problemlos auf Humangewebe übertragen werden konnte. Die Quantifizierung von Ki67 positiven Zellen erfolgte entlang der Körnerzellschicht in einem definierten Abstand in der Einheit Zellen pro Millimeter. Die Ergebnisse der hier vorliegenden Studie widersprechen in mehrfacher Hinsicht den Hypothesen, die sich aus tierexperimentellen Studien ergeben. Während die neurale Stammzellproli-feration bei schizophrenen Psychosen signifikant vermindert ist, findet sich kein Unterschied bei affektiven Erkrankungen im Vergleich zu Kontrollen. Weder wird die „Neurogenese-Hypothese“ der Depression bestätigt, noch zeigte sich ein Effekt antidepressiv oder antipsychotisch wirksamer Pharmaka auf die Rate adulter Neurogenese, da eine pharmakologische Therapie jedweder Art keinen Einfluss auf die Zahl Ki67 positiver Zellen hatte. Deshalb scheint eine Steigerung der adulten Neurogenese kein Wirkmechanismus dieser Medikamente zu sein. Ein überraschendes Ergebnis jedoch ist die signifikant reduzierte Rate adulter Neurogenese bei an Schizophrenie erkrankten Patienten. Aufgrund der sehr begrenzten Anzahl untersuchter Patienten ist die vorliegende Studie in ihrer Aussagekraft jedoch eingeschränkt und muss daher an einem größeren Patientenkollektiv wiederholt werden. Eine Vielzahl von Fragen bzgl. des Stellenwerts der adulten Neurogenese bei psychischen Erkrankungen bleibt darüber hinaus weiter ungeklärt, was die Durchführung weiterer Studien am adulten humanen Gehirn verlangt. / The phenomenon of adult neurogenesis (AN), that is, the generation of functional neurons from neural stem cells in the dentate gyrus of the hippocampus, has attracted remarkable attention, especially as it was shown that this process is also active in the human brain. Based on animal studies, it has been suggested that reduced AN is implicated in the etiopathology of psychiatric disorders, and that stimulation of AN contributes to the mechanism of action of antidepressant therapies. As data from human post-mortem brain are still lacking, we investigated whether the first step of AN, that is, the level of neural stem cell proliferation (NSP; as quantified by Ki-67 immunohistochemistry), is altered in tissue from the Stanley Foundation Neuropathology Consortium comprising brain specimens from patients with bipolar affective disorder, major depression, schizophrenia as well as control subjects (n = 15 in each group). The hypothesis was that stem cell proliferation is reduced in affective disorders, and that antidepressant treatment increases NSP. Neither age, brain weight or pH, brain hemisphere investigated nor duration of storage had an effect on NSP. Only in bipolar disorder, postmortem interval was a significant intervening variable. In disease, onset of the disorder and its duration likewise did not affect NSP. Also, cumulative lifetime dose of fluphenazine was not correlated with NSP, and presence of antidepressant treatment did not result in an increase of NSP. Concerning the different diagnostic entities, reduced amounts of newly formed cells were found in schizophrenia, but not in major depression. Our findings suggest that reduced NSP may contribute to the pathogenesis of schizophrenia, whereas the rate of NSP does not seem to be critical to the etiopathology of affective disorders, nor is it modified by antidepressant drug treatment.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-05-17T21:14:35Z No. of bitstreams: 1 AnaGiseliaNascimento.pdf: 3146962 bytes, checksum: 2e04b37893415a3b55c8a3bcd2cf19b9 (MD5) / Made available in DSpace on 2017-05-17T21:14:35Z (GMT). No. of bitstreams: 1 AnaGiseliaNascimento.pdf: 3146962 bytes, checksum: 2e04b37893415a3b55c8a3bcd2cf19b9 (MD5) Previous issue date: 2016-09-30 / Pituitary adenomas represent 10-15% of all intracranial tumors with an incidence of 1/1000 of the population. They are considered benign and mostly slow growth. Despite this, about 35% may present with invasion of parasellar compartments and a significant number have a clinically aggressive behavior with early recurrence during follow-up, despite surgery and other therapeutic approaches. Such aggressive tumors should be identified early in order to provide a more incisive treatment and / or a strict clinical and radiological follow-up, with prompt djunctive therapy institution in persistence or recurrence of these tumors. Despite these considerations, to date, no clinical, radiological and histological marker predicting such behavior has been established. This study aims to reassess the role of Ki-67 through an improved immunohistochemical technique for the expression of these antigens and correlate it to demographic and clinicopathological parameters of pituitary adenomas. This is a crosssectional study conducted from November 2015 to August 2016 were used pituitary adenoma tissue from 62 patients from the Endocrinology Service of the University of the Federal University of Maranhão Hospital. For evaluation of adenomas of medical records data were analyzed, namely: clinical and serum levels of related hormones as well as magnetic resonance imaging of the pituitary gland. To analyze the expression of Ki-67 was performed immunohistochemical technique using the anti-human monoclonal primary antibody, expressed by the percentage of labeled nuclei in the form of labeling index Ki-67. In our sample of 62 patients, 31 patients were female (50%) with a mean age at surgery of 46.2 (± 13.3) years, ranging from 16 to 73 years. Ki-67 index, as well as their median had to be higher in females, with p = 0.04 and p = 0.02, respectively. There was no predominance of expression and Ki-67 index with respect to age and diagnosis. Adenomas giant had higher median Ki-67 with respect to non giants with p = 0.04, but there was no correlation of Ki-67 with tumor invasion. The pituitary adenomas previously submitted to specific pharmacotherapy showed expression of Ki-67 lower, however, only when the somatotropinomas were analyzed separately, there was statistically significant with p = .0,03. The Ki-67 antigen, a marker of tumor proliferative activity, play an important role in pituitary adenomas. In conjunction with other biomarkers such as tumor invasion, large volume adenoma the initial presentation of patients and / or rapid tumor growth rate documentation can lead to identification of patients which show an aggressive clinical behavior, in order to conduct a multimodal therapeutic approach more effective or more strict postoperative follow-up. / Adenomas hipofisários representam de 10-15% de todas as neoplasias intracranianas com prevalência de um caso para 1000 habitantes. Apresentam grande variação de comportamento clínico, sendo a maioria de crescimento indolente, enquanto cerca de 35% podem ser invasivos e um pequeno grupo francamente agressivo. Até o presente momento, não há um biomarcador que possa predizer seu comportamento de forma confiável, o que possibilitaria uma terapia adjuvante mais agressiva ou um seguimento clínico mais rigoroso. O objetivo desse estudo foi avaliar a expressão e o índice do marcador imuno-histoquímico de proliferação celular, antígeno Ki-67 em adenomas hipofisários e correlacioná-lo a parâmetros demográficos e clínico-patológicos, visando definir sua relevância prognóstica. Trata-se de um estudo transversal realizado no período de novembro de 2015 a agosto de 2016. Foram utilizados tecidos de adenomas hipofisários de 62 pacientes oriundos do Serviço de Endocrinologia do Hospital Universitário da Universidade Federal do Maranhão. Para avaliação dos adenomas foram analisados dados dos prontuários, a saber: avaliação clínica e dosagens séricas de hormônios relacionados, assim como ressonância nuclear magnética de hipófise. Para analisar a expressão do Ki-67, foi realizada a técnica de imuno-histoquimica utilizando-se o anticorpo primário monoclonal anti-humano expressado pela percentagem de núcleos imunopositivos sob forma de índice de marcação de Ki-67. Na amostra de 62 pacientes, 31 pacientes foram do sexo feminino (50%) com a média de idade à época da cirurgia de 46,2 (±13,3) anos, variando de 16 a 73 anos. O antígeno Ki-67 apresentou expressão em 37 (59,7%) pacientes, variando o seu índice de 0,1% a 2,4%. O índice Ki-67, assim como a sua mediana apresentaram-se mais elevados no sexo feminino com p=0,04 e p=0,02, respectivamente. Não houve predominância da expressão e do índice de Ki-67 com relação a idade e diagnóstico. Adenomas gigantes apresentaram mediana de Ki-67 mais elevada com relação aos não gigantes com valor de p=0,04, mas não houve qualquer correlação de Ki-67 com a invasão tumoral. Os adenomas hipofisários previamente submetidos à farmacoterapia específica apresentaram expressão de Ki-67 mais baixos, no entanto, somente quando os somatotropinomas foram analisados de forma isolada, houve significância estatística com p=0,03. O antígeno Ki-67, marcador de atividade proliferativa tumoral, tem papel relevante em adenomas hipofisários. Em conjunto com outros biomarcadores, como invasão tumoral, grande volume do adenoma à apresentação inicial dos pacientes e/ou documentação de rápida velocidade de crescimento do tumor, pode levar a identificação de pacientes que demonstrem um comportamento clínico agressivo, de forma a conduzir a uma abordagem terapêutica multimodal mais efetiva ou a um seguimento pós operatório mais estrito.

Breast cancer is one of the most common malignancies in women. Due to early detection and the use of screening programs approximately 60% of all new cases lack lymph node involvement. Today, a substantial proportion of these women will be offered adjuvant systemic chemotherapy. However, better proliferation markers are needed to predict patient outcome and to avoid overtreatment. Cyclin A, cyclin E and Ki-67 are all markers for proliferation and involved in the regulation of the cell cycle. Overexpression has been associated with disease recurrence in several studies, but the results have not been consistent. However, none of these studies has investigated aberrant expression of cyclin E (the expression of cyclin E during phases of the cell cycle other than late G1 and early S-phase). Studies have shown that aberrant cyclin E might provide additional prognostic information compared to cyclin E alone. The aims of this thesis were 1.to investigate the prognostic value of cyclin A, cyclin E and aberrant cyclin E in early breast cancer. 2.to validate the tissue microarray (TMA) technique for cyclin A and 3.to define the most optimal cut-off values for cyclin A and Ki-67. We found that the agreement of TMA and large section results was good with kappa values 0.62-0.75 and that the reproducibility of the two readers’ results was good or even very good, with kappa values 0.71 – 0.87. The optimal cut-off value for cyclin A average was 8% and for cyclin A maximum value 11%. The corresponding values for Ki-67 were 15 and 22%. Neither cyclin E nor aberrant cyclin E was a prognostic factor in low-risk node negative breast cancer patients. Finally, we conclude that cyclin A is a prognostic factor in node negative breast cancer (univariate analysis average value OR=2.9 95% CI 1.8-4.6; maximum value OR=3.7 95% CI 2.3-5.9).

Purpose: To determine the expressions of bcl-2, p53, Ki-67 and PTEN on the basis of immunohistochemistry methods in upper urinary transitional cell carcinoma (TCC), and to correlate their presentations in specimens with clinical tumor stage, grade and patient survival. Material and Method: Paraffin-embedded primary upper urinary TCC specimens were divided into 2 groups for immunohistochemical study: Group 1 including 91 cases were treated with bcl-2, p53 and Ki-67 antibodies; group 2 including 93 specimens contained both tumor and benign tissues were treated with PTEN antibody. Semi- quantitatively, according to the amount of the stained cells, they were divided into 3 levels: level 1, scanty; level 2, focal; and level 3, diffuse. Association of immunoreactivity with tumor grade and stage was examined. Prognostic significance of tumor marker expression in patients¡¦ survival was accessed. Results: Group1: Of the 91 tumors most (98.9%) of the specimens showed level 1 bcl-2 expression and only 1 patient had level 2 expression. The p53 mutations were identified level 3 expression in 48.4% of the cases, followed by level 2 (26.4%) and level 1 (25.3%) identifications. The Ki-67 expression was recognized level 3 in 6 patients, level 2 in 21 and level 1 in 66 cases. Significant correlations were seen between p53 expression and tumor grading (p=0.004) and between immunostain of Ki-67 and clinical stage (p=0.031). The p53, bcl-2 and Ki-67 expressions in upper urinary tract TCC specimens were not a significant factor of patients¡¦ survival. Group 2: Of the tumors all cytoplasm has level 3 PTEN expressions and the nuclei, 18 (19.4%) showed scanty expression, 35 (37.6%) revealed focal expression, and diffuse expression was noted in 40 (43.0%) cases. Loss of PTEN expression in tumor nuclei was positively correlated with pathologic stage (p=0.019). Of the fibrocytes adjacent to tumor cells, the nuclei showed 24 (25.8%) scanty, 59 (63.4%) focal and 10 (10.8%) diffuse distribution of PTEN expressions. Poorly differentiated tumor (grade 3) specimens were correlated with loss of PTEN expression in fibrocytic nuclei adjacent to tumor (p=0.028). Most (58%) fibrocytic cytoplasm was scanty PTEN expression, followed by 23 (24.7%) diffuse and 16 (17.2%) focal immunostaining. PTEN expressions in upper urinary tract TCC specimens were not a significant factor of patients¡¦ survival. Conclusions: We examined 93 surgical specimens of upper urinary tract TCC for the expression of PTEN and 91 cases for bcl-2, p53 and Ki-67 by immunohistochemical stained. Correlation between tumor grading and p53 mutations and correlation between clinical stage and Ki-67 immunoreactivity were observed. Meanwhile, loss of PTEN expression in tumor nuclei of upper urinary TCC is correlated significantly with advanced tumor stage, and poorly differentiated tumor specimens were correlated with loss of PTEN expression in normal nuclei adjacent to tumor cells. However, no correlation between overall survival rate and tumor markers was identified. Thus, the detection of p53, bcl-2, Ki-67 and PTEN would be not enough for evaluation the prognosis of upper TCC.

O propósito do presente estudo foi analisar as células epiteliais odontogênicas procurando um entendimento maior sobre a natureza e conseqüentemente o comportamento de algumas lesões odontogênicas. A expressão imunoistoquímica de p53 e PCNA foi analisada em cisto radicular, cisto dentígero, ceratocisto odontogênico e cisto odontogênico calcificante (Cisto de Gorlin) onde verificou-se que no cisto radicular e cisto dentígero a expressão dos marcadores está relacionado com proliferação e stress celular causado pelo estímulo inflamatório e em ceratocisto odontogênico e Cisto de Gorlin a expressão dos marcadores corresponde a proliferação celular não descartando também a presença de mutação no gene TP53. Também foi observada a expressão de Ki-67, EGFR e Survivin em folículo pericoronário, ceratocisto odontogênico e cisto dentígero que mostrou que as células epiteliais dos folículos pericoronários têm potencial proliferativo para formar lesões odontogênicas e que a proliferação das células do cisto dentígero é relacionada com o estímulo inflamatório. Todos os marcadores estudados comprovaram a natureza neoplásica do ceratocisto odontogênico. / The purpose of this study was to analyze odontogenic epithelial cells to contribute to the knowledge about their nature and, consequently, about the behavior of certain odontogenic lesions. Immunohistochemical expressions of p53 and PCNA were analyzed in radicular cysts, dentigerous cysts, odontogenic keratocysts and calcifying odontogenic cysts (Gorlin cyst). In radicular and dentigerous cysts, the expression of these markers was associated with cell proliferation and stress caused by an inflammatory stimulus. In keratocysts and Gorlin cysts, the expression of markers corresponded to cell proliferation. Results also showed possible mutation in TP53 gene. Also, Ki-67, EGFR and Survivin were expressed in pericoronal follicles, odontogenic keratocysts and dentigerous cysts, which demonstrated that epithelial cells of pericoronal follicles may proliferate to form odontogenic lesions and that cell proliferation in dentigerous cysts was associated with an inflammatory stimulus. The analysis of all markers under study confirmed the neoplastic nature of odontogenic keratocysts.

Resumo não disponível.