Os autores realizaram uma avaliação de 246 casos de meningeomas diagnosticados no Departamento de Patologia da Fundação Faculdade Federal de Ciências Médicas de Porto Alegre durante 25 anos (março de 1968 a março de 1993). Esses tumores foram classificados como típico, atípico, anaplásico e papilar segundo critérios previamente definidos. Desses casos foram separados, aleatoriamente, 116 tumores (60 típicos, 46 atípicos, 9 anaplásicos e 1 papilar) e realizada técnica imuno-histoquímica para receptores de estrógeno e progesterona com o objetivo de determinar se existe diferença entre tumores típicos e não típicos em relação aos receptores hormonais. Entre os critérios utilizados para definição de tumores não típicos, a invasão do SNC predominou com 62,3% dos casos. Áreas de necrose estiveram presentes em 45,9% dos meningeomas, 36% apresentaram aumento do número de mitoses, 39,3% foram hipercelular e 32,8% mostraram anaplasia citológica. Os meningeomas foram típicos em 75,22% dos casos, atípicos em 19,1%, anaplásicos em 4,47% e papilar em 1,21%. A localização supratentorial foi mais freqüente em ambos os grupos (típicos e não típicos) e houve predomínio pelo sexo feminino. A faixa etária mais acometida foi entre 51 e 60 anos. Houve recidiva em 3,78% dos meningeomas típicos, em 42,55% dos atípicos, 45,45% dos anaplásicos e não houve recidiva nos 3 casos de tumores com características papilares. A técnica imuno-histoquímica para receptores de estrógeno foi negativa em todos os meningeomas estudados. Os receptores de progesterona foram detectados, pela imuno-histoquímica, em 58,33% dos meningeomas típicos e em 48,21% dos tumores não típicos. Essa diferença não foi estatisticamente significativa. Entretanto, considerando individualmente os critérios utilizados para seleção dos não típicos, os tumores que apresentaram, de forma concomitante, invasão do SNC e aumento da taxa mitótica ou necrose, bem como a soma das três característica, foram predominantemente negativos para receptor de progesterona (p=0,038; p=0,001 e p=0,044 respectivamente). Os autores concluem que os critérios utilizados para definição de típicos e não típicos foram adequados para predizer maior chance de recidiva tumoral; que os receptores de estrógeno não estão presentes em meningeomas; que receptores de progesterona estão presentes na maioria dos meningeomas estudados; que a presença de receptor de progesterona isoladamente não é suficiente para predizer maior malignidade tumoral; e que nos tumores que apresentam invasão do SNC associado a áreas de necrose e/ou aumento da taxa mitótica, predominaram os negativos para receptor de progesterona, fazendo crer que esse grupo deve apresentar resposta pobre a uma possível manipulação hormonal. / The authors assessed 246 cases of meningiomas, diagnosed in the Pathology Department of Fundação Faculdade Federal de Ciências Médicas de Porto Alegre during 25 years (from March 1968 to March 1993). Those tumors were classífied as typical, atypical, anaplastic, and papillary, according to previously defined criteria. From this cases, 116 tumours (60 typical, 46 atypical, 9 anaplastic and 1 papillary) were randomiy selected, being used an immunohistochemical technique for estrogen and progesterone receptors, attempting to determine if there is any difference between typical and non-typical tumours, in relation to hormone receptors. Among the criteria used to define non-typical tumours, brain invasion was predominant in 62,3%. There were areas of necrosis in 45,9% of meningiomas, 36% displayed increased mitotic activity, 39,3% were hypercellular, and 32,8% cytological anaplasia. Meningiomas were typical in 75,22%, atypical in 19,1%, anaplastic in 4,47%, and papillary in 1,21% of the cases. Supratentorial location was more frequent in both groups (typical and nontypical), and female gender predominated. The most afflicted age group was between 51 and 60 years of age. There was recurrence in 3,78% of typical. 42,55% of atypical. 45,45% of anaplastic, and there was no recurrence in those tumours with papillary fegtures. The immunohistochemical technique to estrogen receptors was negative in ali meningiomas studied. Progesterone receptors were detected by immunohistochemstry in 58,33% of typical, and in "48,21% of non-typicai meningiomas. This difference was not statisticaliy significant. However, individually considering the criteria used for selection of non-typical tumours, those that concurrently displayed brain invasion and increased mitotic activity or necrosis, as well as the summation of those three features, were predominantiy negative for progesterone receptors (respectively p=0,038; p=0,001; and p=0,044). The authors conclusion was that the criteria used to define typical and nontypical were adequate to predict a higher chance of tumour recurrence, that estrogen receptor were not present in meningiomas; that progesterone receptors in isolation is not enough to predict a higher tumoral malignancy; and in tumours that showed brain invasion. associated to necrosis and/or increased mitotic activity, there was a predominance of negatives to progesterone receptors, infering this group should display a poor response to a possible hormonal manipulation.

Objetivo: Identificar los factores relacionados con la recurrencia de los meningiomas intracraneales operados en el Instituto Especializado de Enfermedades Neoplásicas “Eduardo Cáceres Graziani”. Material y Métodos: Estudio retrospectivo, comparativo, analítico, de caso control. La población de estudio estuvo constituida por los pacientes operados con diagnóstico anatomopatológico de meningioma intracraneal que fueron operados desde el año 1984 a 1997, con control postoperatorio a cinco años. Resultados: 55 pacientes cumplieron los criterios de inclusión; de estos, 23 (42%) presentaron recurrencia y 32 (58%) no presentaron recurrencia. Se realizó la remoción quirúrgica total en 36 pacientes y subtotal en 19 pacientes. Tuvieron recurrencia el 57.8% de los pacientes con resección quirúrgica subtotal y el 33.3% de pacientes con remoción total; esto nos da un OR de 2,7. con una p< 0.05. Con respecto al sexo, se observó mayor recurrencia del meningioma en pacientes de sexo masculino, con un OR de 1.7. La edad promedio de los pacientes con recurrencia fue de 43.9 ± 16.3 años versus 40. 85 ±17.7 del grupo control. El tiempo de recurrencia fue en promedio de 30.7 ± 16.3 meses. La localización del meningioma que más recurre fue la zona temporal, relacionada con la base de cráneo y con el ala mayor del esfenoides. El abordaje que presentó mayor recurrencia fue el frontoparietal, mientras que el frontal, parietal, frontoparietotemporal y el temporoparietal tienen menor recurrencia. Con respecto al tipo histológico no existe preponderancia de ningún tipo de meningioma con respecto a la recurrencia. Conclusiones: El factor más importante relacionado con la recurrencia del meningioma intracraneal operado fue el grado de resección quirúrgica, encontrándose que un paciente sometido a una remoción quirúrgica subtotal tiene 2,7 veces más posibilidades de presentar recurrencia del meningioma en comparación con la resección quirúrgica total. / --- Objective: To identify the factors related to the recurrence of meningioma intracraneal patient in the Specialized Institute of Neoplasicas Diseases. “Eduardo Cáceres Graziani”. Material and Methods: Retrospective, comparative, analytical study, of case control. The study population was constituted by patients with intracraneal diagnosis of meningioma patients from year 1984 to 1997, with postoperating control to five years. Results: 55 patients fulfilled the inclusion criteria; of these, 23 (42%) presented recurrence and 32 (58%) did not present recurrence. It was made total the surgical removal in 36 patients and subtotal in 19 patients. The 57,8% of the patients with subtotal surgical removal and the 33,3% of patients with total removal had recurrence; this gives a OR us of 2.7, with p< 0.05.With respect to sex, greater recurrence of meningioma in patients of masculine sex was observed, with a OR of 1.7.The age average of the patients with recurrence is of 43,9 ± 16,3 years versus 40. 85 ±17.7 of the group control. The time of recurrence ± was in average of 30,7 16,3 months. The location of meningioma that resorts more is the temporary zone, related to the base of skull and the greater wing of the esfenoides. The boarding that presents greater recurrence was the frontoparietal, whereas the frontal, parietal, frontoparietotemporal and the temporoparietal have minor recurrence. With respect to the histologic type superiority of any type of meningioma with respect to the recurrence does not exist. Conclusions: The most important factor in the recurrence of meningioma was the degree of surgical resection, being that a patient submissive subtotal a surgical removal has 2.7 times more possibilities of presenting recurrence of meningioma. / Tesis de segunda especialidad

Meningiomas are the most common primary brain tumors, yet few successful non-invasive treatment options are available for patients. Immunotherapy has provided new insights into treatments for solid tumors. The immune infiltrate of meningiomas has been broadly characterized, showing a significant monocytic cell content, but little is known about the phenotype and function of these myeloid cells within the tumor environment. As circulating monocytes differentiate into macrophages with highly plastic character within tissue, it remains to be seen how the macrophages in meningiomas are influencing the tumor. As many studies have described the presence of monocytic subpopulations within other solid tumors, we hypothesize that meningiomas contain two populations of myeloid cells: a pro-inflammatory macrophage-like population, and an immunosuppressive myeloid-derived suppressor cell (MDSC)-like population. We collected fresh tumor samples and processed them into a single-cell suspension. The cells were then stained with fluorescently labeled surface marker antibodies commonly found on macrophages and MDSCs. We used flow cytometry to quantify the myeloid populations, sorted the populations with a FACSAria™, and analyzed their gene expression profiles with NanoString® and TaqMan®. Two distinct myeloid populations were found in all analyzed tumor samples, varying in macrophage-like to MDSC-like ratios from tumor to tumor. Gene expression analysis of these populations confirmed the sorting strategy and provided new clues into the identity and function of the myeloid cell populations infiltrating meningiomas. NanoString® results confirmed a high HLA-DR gene expression in the HLA-DR+ sorted populations. The tumor HLA-DR+ population was found to have higher gene expression relative to the HLA-DR- population for chemoattractants such as IL8, CCL3, and CCL4. Compared to a healthy blood monocyte control, tumor myeloid cells expressed higher levels of the genes C3AR1, ROCK2, IL10, NOS2, IL18, and CSF2. Finally, qPCR analysis and NanoString® results showed high expression of the gene IL6 in the non-immune cell tumor cells (CD45-). The IL6 cytokine has been shown to induce MDSCs. These findings may have significant implications in identifying new targets in immunotherapy to stop tumor growth and increase survival outcomes. / 2016-11-03T00:00:00Z

Introdução: A capacidade funcional refere-se fundamentalmente à potencialidade humana para o desempenho ocupacional, imprescindível para uma melhor qualidade de vida. O objetivo desta pesquisa, aprovada pelo Comitê de Ética em Pesquisa do HCFMRP-USP (processo no 648/2008), foi avaliar a capacidade funcional de pacientes submetidos a neurocirurgia oncológica e sua relação com a qualidade de vida. Método: Durante o período de maio/08 a abril/09 (12 meses) foram avaliados 52 sujeitos adultos de ambos os sexos; o grupo experimental teve 26 pacientes, sendo 14 com diagnóstico de meningioma (grupo1) e 12 com diagnóstico de glioma de alto grau (grupo 2); e o grupo controle teve 26 sujeitos, subdivididos em grupos: 1A (pareado com o grupo 1) e 2A (pareado com grupo 2). Foram realizados dois tipos de dinamometria - avaliação de força de preensão (Grip TrackTM Testing) e de pinça (Pinch TrackTM Testing) com equipamento computadorizado Tracker SystemTM; aplicação de protocolos internacionais validados no Brasil - Hospital Anxiety and Depression Scale HAD, Item Short-Form Health Survey - SF-36 e Health Assessment Questionnaire HAQ, no período pré-operatório e no terceiro mês de pós-operatório. Na análise estatística foram aplicados os testes Mann-Whitney, Wilcoxon e Coeficiente de Correlação de Spearman Resultados: Constatou-se que nos três primeiros meses de pós operatório houve aumento da capacidade funcional dos sujeitos dos grupos experimentais (1 e 2) e diminuição dos sintomas de ansiedade e depressão; não houve diferença significativa nos testes de força no pré e no pós operatório, mas ambos os grupos apresentam diminuição de força no membro dominante em comparação com os grupos controle. Houve correlações de forte magnitude entre os dados coletados através dos testes Grip TrackTM Testing, Pinch TrackTM Testing; protocolos: HAD, HAQ e SF-36, com rho 0,600 a 0,969. Os domínios aspecto social e emocional do instrumento de qualidade de vida mostraram uma piora no pósoperatório imediato. Esses resultados corroboram a compreensão ampliada do conceito de funcionalidade, como proposto na Classificação Internacional de Funcionalidade e Incapacidade e Saúde\" (CIF). Conclusão: A funcionalidade está diretamente relacionada com a qualidade de vida, principalmente nos aspectos psicossociais, sendo necessária compreendê-la de forma mais ampla do que funções físicas específicas, para implementar planos de tratamentos mais adequados para os sujeitos com tumor cerebral, tanto no pré como no pós operatório, com acompanhamento de uma equipe multidisciplinar. / Introduction. Functional capacity refers mainly to the potential for human occupational performance, essential for a better quality of life. This research, approved by the Committee of Ethics in Research- HCFMRP USP (in case 648/2008), was to assess the functional capacity of patients undergoing neurosurgery oncology and its relationship to quality of life. Methods. During the period of the abril/09 May/08 (12 months) were assessed 52 subjects adults of both sexes, the experimental group had 26 patients, 14 with a diagnosis of meningioma (group1) and 12 diagnosed with high grade glioma (group 2); and the control group has 26 subjects, divided into groups: 1A (paired with group 1) and 2A (paired with group 2). Were performed two types of dinamometry - evaluation of grip strength (grip TrackTM Testing) and pinch (Pinch TrackTM Testing) SystemTM Tracker with computerized equipment, application of international protocols validated in Brazil - Hospital Anxiety and Depression Scale - HAD, Item Short- Form Health Survey - SF-36 and Health Assessment Questionnaire - HAQ in the preoperative period and in the third month postoperatively. Statistical analysis using the statistical Mann-Whitney, Wilcoxon and Spearman Correlation Coefficient. Results . It was found that the first three months postoperatively increased functional capacity of the subjects of the experimental groups (1 and 2) and decrease symptoms of anxiety and depression, no significant difference in the strength tests before and after surgery but both groups have reduced strength in the dominant limb compared with control groups. There were strong correlations between the magnitude of data collected through tests Grip TrackTM Testing, Testing Pinch TrackTM; protocols: HAD, HAQ and SF-36 with rho 0.600 to 0.969. The aspect of social and emotional domains of quality of life instrument showed a worsening in the immediate postoperative period. These results support the expanded understanding of the concept of functionality, as proposed in the \"International Classification of Functioning and Disability and Health (ICF). Conclusion. The functionality is directly related to the quality of life, especially on psychosocial aspects and needed to understand it more broadly than specific physical functions, to implement plans of treatment best suited to individuals with brain tumor, both pre and postoperatively, with the accompaniment of a multidisciplinary team.

Introdução: A capacidade funcional refere-se fundamentalmente à potencialidade humana para o desempenho ocupacional, imprescindível para uma melhor qualidade de vida. O objetivo desta pesquisa, aprovada pelo Comitê de Ética em Pesquisa do HCFMRP-USP (processo no 648/2008), foi avaliar a capacidade funcional de pacientes submetidos a neurocirurgia oncológica e sua relação com a qualidade de vida. Método: Durante o período de maio/08 a abril/09 (12 meses) foram avaliados 52 sujeitos adultos de ambos os sexos; o grupo experimental teve 26 pacientes, sendo 14 com diagnóstico de meningioma (grupo1) e 12 com diagnóstico de glioma de alto grau (grupo 2); e o grupo controle teve 26 sujeitos, subdivididos em grupos: 1A (pareado com o grupo 1) e 2A (pareado com grupo 2). Foram realizados dois tipos de dinamometria - avaliação de força de preensão (Grip TrackTM Testing) e de pinça (Pinch TrackTM Testing) com equipamento computadorizado Tracker SystemTM; aplicação de protocolos internacionais validados no Brasil - Hospital Anxiety and Depression Scale HAD, Item Short-Form Health Survey - SF-36 e Health Assessment Questionnaire HAQ, no período pré-operatório e no terceiro mês de pós-operatório. Na análise estatística foram aplicados os testes Mann-Whitney, Wilcoxon e Coeficiente de Correlação de Spearman Resultados: Constatou-se que nos três primeiros meses de pós operatório houve aumento da capacidade funcional dos sujeitos dos grupos experimentais (1 e 2) e diminuição dos sintomas de ansiedade e depressão; não houve diferença significativa nos testes de força no pré e no pós operatório, mas ambos os grupos apresentam diminuição de força no membro dominante em comparação com os grupos controle. Houve correlações de forte magnitude entre os dados coletados através dos testes Grip TrackTM Testing, Pinch TrackTM Testing; protocolos: HAD, HAQ e SF-36, com rho 0,600 a 0,969. Os domínios aspecto social e emocional do instrumento de qualidade de vida mostraram uma piora no pósoperatório imediato. Esses resultados corroboram a compreensão ampliada do conceito de funcionalidade, como proposto na Classificação Internacional de Funcionalidade e Incapacidade e Saúde\" (CIF). Conclusão: A funcionalidade está diretamente relacionada com a qualidade de vida, principalmente nos aspectos psicossociais, sendo necessária compreendê-la de forma mais ampla do que funções físicas específicas, para implementar planos de tratamentos mais adequados para os sujeitos com tumor cerebral, tanto no pré como no pós operatório, com acompanhamento de uma equipe multidisciplinar. / Introduction. Functional capacity refers mainly to the potential for human occupational performance, essential for a better quality of life. This research, approved by the Committee of Ethics in Research- HCFMRP USP (in case 648/2008), was to assess the functional capacity of patients undergoing neurosurgery oncology and its relationship to quality of life. Methods. During the period of the abril/09 May/08 (12 months) were assessed 52 subjects adults of both sexes, the experimental group had 26 patients, 14 with a diagnosis of meningioma (group1) and 12 diagnosed with high grade glioma (group 2); and the control group has 26 subjects, divided into groups: 1A (paired with group 1) and 2A (paired with group 2). Were performed two types of dinamometry - evaluation of grip strength (grip TrackTM Testing) and pinch (Pinch TrackTM Testing) SystemTM Tracker with computerized equipment, application of international protocols validated in Brazil - Hospital Anxiety and Depression Scale - HAD, Item Short- Form Health Survey - SF-36 and Health Assessment Questionnaire - HAQ in the preoperative period and in the third month postoperatively. Statistical analysis using the statistical Mann-Whitney, Wilcoxon and Spearman Correlation Coefficient. Results . It was found that the first three months postoperatively increased functional capacity of the subjects of the experimental groups (1 and 2) and decrease symptoms of anxiety and depression, no significant difference in the strength tests before and after surgery but both groups have reduced strength in the dominant limb compared with control groups. There were strong correlations between the magnitude of data collected through tests Grip TrackTM Testing, Testing Pinch TrackTM; protocols: HAD, HAQ and SF-36 with rho 0.600 to 0.969. The aspect of social and emotional domains of quality of life instrument showed a worsening in the immediate postoperative period. These results support the expanded understanding of the concept of functionality, as proposed in the \"International Classification of Functioning and Disability and Health (ICF). Conclusion. The functionality is directly related to the quality of life, especially on psychosocial aspects and needed to understand it more broadly than specific physical functions, to implement plans of treatment best suited to individuals with brain tumor, both pre and postoperatively, with the accompaniment of a multidisciplinary team.

by Jenney Yin-mei Tse. / Thesis (Ph.D.)--Chinese University of Hong Kong, 1997. / Includes bibliographical references (p. 122-146). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web.

Glioblastoma multiforme (GBM) is the most common primary brain tumor. Given the current standard of care, the prognosis for patients diagnosed with this disease is still poor. There consequently exists a need to improve current treatments, as well as to develop new ones. Many obstacles however need to be overcome to facilitate this effort and one of these involves the development of improved methods to monitor treatment effects. At present, the effects of treatment are typically assessed by radiological means several months after its initiation, which is unsatisfactory for a fast growing tumor like GBM. It is however likely that treatment effects can be detected on a molecular level long before radiological response, especially considering many of the targeted therapies that are currently being developed. Biomarkers for treatment efficacy may be of great importance in the future individualization of brain tumor treatment. The work presented herein was primarily focused on detecting early effects of GBM treatment. To this end, we designed experiments in the BT4C rat glioma model in which we studied effects of both conventional radiotherapy and an experimental angiogenesis inhibitor, vandetanib. Brain tissue samples were analyzed using a high throughput mass spectrometry (MS) based screening, known as Surface Enhanced Laser Desorption/Ionization - Time of Flight - Mass Spectrometry (SELDI-TOF-MS). The vast amounts of data generated were subsequently analyzed by established multivariate statistical methods, such as Principal Component Analysis (PCA), Partial Least Squares (PLS), and Orthogonal Partial Least Squares (OPLS), developed for analysis of large and complex datasets. In the radiotherapy study we detected a protein spectrum pattern clearly related to tumor progression. We notably observed how this progression pattern was hampered by radiotherapy. The vandetanib study also revealed significant alterations of protein expression following treatment of different durations, both in tumor tissue and in normal brain contralateral to the tumor. In an effort to further elucidate the pathophysiology of GBM, particularly in relation to treatment, we collected extracellular fluid (ECF) samples from 11 patients diagnosed with inoperable GBM. The samples were collected by means of stereotactic microdialysis, both from within the contrast enhancing tumor and the brain adjacent to tumor (BAT). Samples were collected longitudinally from each patient in a time span of up to two weeks, during which the patient received the first five fractions of radiotherapy. The ECF samples were then analyzed by Gas Chromatography Mass Spectrometry (GC-MS) to screen them with respect to concentrations of low molecular weight compounds (metabolites). Suitable multivariate analysis strategies enabled us to extract patterns of varying metabolite concentrations distinguishing between samples collected at different locations in the brain as well as between samples collected at different time points in relation to treatment. In a separate study, we also applied SELDI-TOF-MS and multivariate statistical methods to unravel possible differences in protein spectra between invasive and non-invasive WHO grade I meningiomas. This type of tumor can usually be cured by surgical resection however sometimes it grows invasively into the bone, ultimately causing clinical problems. This study revealed the possibility to differentiate between invasive and non-invasive benign meningioma based on the expression pattern of a few proteins. Our approach, which includes sample analysis and data handling, is applicable to a wide range of screening studies. In this work we demonstrated that the combination of MS screening and multivariate analyses is a powerful tool in the search for patterns related to treatment effects and diagnostics in brain tumors.

The WHO classification of brain tumors is based on histological features and the aggressiveness of the tumor is classified from grade I to IV, where grade IV is the most aggressive. Today, the correlation between prognosis and tumor grade is the most important component in tumor classification. High grade gliomas, glioblastomas, are associated with poor prognosis and a median survival of 14 months including all available treatments. Low grade meningiomas, usually benign grade I tumors, are in most cases cured by surgical resection. However despite their benign appearance grade I meningiomas can, without any histopathological signs, in some cases develop bone invasive growth and become lethal. Thus, it is necessary to improve conventional treatment modalities, develop new treatment strategies and improve the knowledge regarding the basic pathophysiology in the classification and treatment of brain tumors. In this thesis, both proteomics and metabolomics have been applied in the search for biomarkers or biomarker patterns in two different types of brain tumors, gliomas and meningiomas. Proteomic studies were carried out mainly by surface enhanced laser desorption ionization time of flight mass spectrometry (SELDI-TOF-MS). In one of the studies, isobaric tags for relative and absolute quantitation (iTRAQ) labeling in combination with high-performance liquid chromatography (HPLC) was used for protein detection and identification. For metabolomics, gas-chromatography time-of-flight mass spectrometry (GC-TOF-MS) has been the main platform used throughout this work for generation of robust global metabolite profiles in tissue, blood and cell cultures. To deal with the complexity of the generated data, and to be able to extract relevant biomarker patters or latent biomarkers, for interpretation, prediction and prognosis, bioinformatic strategies based on chemometrics were applied throughout the studies of the thesis. In summary, we detected differentiating protein profiles between invasive and non-invasive meningiomas, in both fibrous and meningothelial tumors. Furthermore, in a different study we discovered treatment induce protein pattern changes in a rat glioma model treated with an angiogenesis inhibitor. We identified a cluster of proteins linked to angiogenesis. One of those proteins, HSP90, was found elevated in relation to treatment in tumors, following ELISA validation. An interesting observation in a separate study was that it was possible to detect metabolite pattern changes in the serum metabolome, as an effect of treatment with radiotherapy, and that these pattern changes differed between different patients, highlighting a possibility for monitoring individual treatment response.  In the fourth study of this work, we investigated tissue and serum from glioma patients that revealed differences in the metabolome between glioblastoma and oligodendroglioma, as well as between oligodendroglioma grade II and grade III. In addition, we discovered metabolite patterns associated to survival in both glioblastoma and oligodendroglioma. In our final work, we identified metabolite pattern differences between cell lines from a subgroup of glioblastomas lacking argininosuccinate synthetase (ASS1) expression, (ASS1 negative glioblastomas), making them auxotrophic for arginine, a metabolite required for tumor growth and proliferation, as compared to glioblastomas with normal ASS1 expression (ASS1 positive). From the identified metabolite pattern differences we could verify the hypothesized alterations in the arginine biosynthetic pathway. We also identified additional interesting metabolites that may provide clues for future diagnostics and treatments. Finally, we were able to verify the specific treatment effect of ASS1 negative cells by means of arginine deprivation on a metabolic level.

Neurofibromatosis type 2 (NF2) is an autosomal dominant inherited condition that predisposes individuals to develop multiple nervous system tumours, primarily schwannoma, meningioma and ependymoma. NF2 is characterised by loss of the tumour suppressor protein Merlin, caused by bi-allelic mutations of the encoding gene NF2 or by loss of heterozygosity. These tumours can occur either sporadically or as part of the NF2 condition. The majority are slow growing and display benign features, but this benignancy renders them largely unresponsive to classic chemotherapeutic agents leaving surgery and radiosurgery as the only remaining treatment options. Depending on their location, NF2-related tumours can cause a number of side effects, including nausea, balance problems, and in some cases hearing and/or vision loss. Phosphorylation is a key regulatory mechanism leading to changes in cell signalling. By identifying phosphoproteins that are significantly activated in tumour cells, novel therapies can be developed aiming to specifically target the phosphorylated status of these proteins thus ‘switching off’ the signalling cascade. The objective of this study is to identify and validate common targets in both Merlin-deficient meningioma and schwannoma to eventually exploit in novel therapeutic approaches. Using phosphoprotein purification followed by mass spectrometry analysis, we identified Signal Transducer and Activator of Transcription 1 (STAT1), phosphorylated at Serine (S) 727 and Tyrosine (Y) 701, PDZ and LIM domain protein 2 (PDLIM2), Heat Shock 70kDa Protein 1A (HSPA1A) and Filamin B (FLNB) as potential common, novel therapeutic targets. We validated these candidates in human primary meningioma and schwannoma tumour cells using a variety of techniques. We also showed that specific 7 knockdown of STAT1 and PDLIM2 was related to a significant decrease in cellular proliferation. Additionally, we performed co-immunoprecipitation using PDLIM2 as the bait protein and identified STAT1, HSPA1A and FLNB as binding partners, suggesting a novel interaction network involving all of the potential targets identified in this study. We also identified activation of several pathways and/ or biological processes in both tumour types that warrant further investigation i.e. endocytosis in schwannoma and the proteasome in meningioma. In conclusion, with our approach we substantially increased the overall body of knowledge regarding the proteome and phosphoproteome of meningioma and schwannoma. We generated a comprehensive set of data that highlighted several potential therapeutic targets and dysregulated pathways which will be further investigated.

Indiana University-Purdue University Indianapolis (IUPUI) / Meningioma represent more than one-third of all primary central nervous system (CNS) tumors, and it can be classified into three grades according to WHO (World Health Organization) in terms of clinical aggressiveness and risk of recurrence. A key component of meningioma grades is the mitotic count, which is defined as quantifying the number of cells in the process of dividing (i.e., undergoing mitosis) at a specific point in time. Currently, mitosis counting is done manually by a pathologist looking at 10 consecutive high-power fields (HPF) on a glass slide under a microscope, which is an extremely laborious and time-consuming process. The goal of this thesis is to investigate the use of computerized methods to automate the detection of mitotic nuclei with limited labeled data. We built computational methods to detect and quantify the histological features of mitotic cells on a whole slides image which mimic the exact process of pathologist workflow. Since we do not have enough training data from meningioma slide, we learned the mitotic cell features through public available breast cancer datasets, and predicted on meingioma slide for accuracy. We use either handcrafted features that capture certain morphological, statistical, or textural attributes of mitoses or features learned with convolutional neural networks (CNN). Hand crafted features are inspired by the domain knowledge, while the data-driven VGG16 models tend to be domain agnostic and attempt to learn additional feature bases that cannot be represented through any of the handcrafted features. Our work on detection of mitotic cells shows 100% recall , 9% precision and 0.17 F1 score. The detection using VGG16 performs with 71% recall, 73% precision, and 0.77 F1 score. Finally, this research of automated image analysis could drastically increase diagnostic efficiency and reduce inter-observer variability and errors in pathology diagnosis, which would allow fewer pathologists to serve more patients while maintaining diagnostic accuracy and precision. And all these methodologies will increasingly transform practice of pathology, allowing it to mature toward a quantitative science.