Background: Multiple sclerosis (MS) is a neurological, autoimmune disease which mainly affects people in the age of 20 to 40. The disease course is unpredictable affecting each patient differently, leading to progressiveand irreversible degradation of the central nervous system. There is no treatment that cures this disease, however, there are treatments that either slows down the disease course or prevents progressive disabilities. A treatment called autologous hematopoietic stem cell transplantation (AHSCT) is thought to reset the immune system and induce a new, more tolerant one, thus haltering the disease course. However, the knowledge about the effects causing the improvement seen in patients treated with AHSCT is limited. Methods: To investigate the effect of AHSCT in MS patients, serum lipidomics data from 16 patients was collected at ten timepoints. The lipidomics data was collected for both positively and negatively charged molecules separately as well as within a single experiment called polarity switching, using mass spectrometry. Since the standard method requires two separate experiments to analyze both positively and negatively charged lipids it requires twice the time and resources compared to polarity switching. Results: Comparing the two mass spectrometry protocols showed that the coefficient of variation (CV) was slightly higher for polarity switching compared to the standard method. Nevertheless, the difference was not significant and both methods had in general a good CV, indicating low technical variation. In addition, this thesis showed that polarity switching has a slightly higher percentage of lipids with zero carryover compared to the standard method. The results also indicated that the expression levels of differentially expressed lipids follow two distinct patterns throughout the AHSCT treatment. The largest intensity variation arises after stem cellreinfusion and the lipid intensities are back to nearly initial levels atthe three month follow-up. Finally, many lipids were found to be associated with the change in c-protein levels as well as erythrocyte, leukocyte, and thrombocyte levels that occurred during treatment. Conclusions: This master thesis showed that polarity switching is a good alternative to the standard method, saving both time and resources without losing too much in specificity. In addition, this thesis has shown that differentially expressed lipids follow two distinct expression patterns through the treatment. The lipids levels for both differentially expressed lipids and lipids associated with clinical data were nearly back to baseline levels three months after AHSCT. Hence, AHSCT has a major but short-lasting impact on the lipid levels in peripheral blood.

A esclerose múltipla (EM) é uma doença desmielinizante inflamatória crônica recorrente, restrita ao sistema nervoso central (SNC), cuja característica histológica é a ocorrência de desmielinização relacionada com infiltrado inflamatório perivenular com relativa preservação axonal. A forma clássica da doença se caracteriza pela recorrência de ataques (surtos), seguidos de remissão dos sintomas (RRMS), com a presença de múltiplas lesões focais dispersas pelo SNC (dissociação espacial) com processo inflamatório exuberante na fase aguda, coexistindo com lesões crônicas (dissociação temporal) sem atividade inflamatória evidenciavel pela quebra de barreira hematoencefálica e realce na fase contrastada na imagem de ressonância magnética (MRI). Alguns pacientes tem uma evolução benigna, permanecendo livre de sequelas significativas por mais de 20 anos da doença. Outros pacientes têm uma forma agressiva da doença, ficando restritos à cadeira de rodas em 8 a 10 anos de evolução. Um desafio é modificar o curso desta forma agressiva, o que pode ser feito com o uso de imunomoduladores, quimioterápicos e, eventualmente, transplante autólogo de células tronco hematopoiéticas (aHSCT). O objetivo da utilização do aHSCT é a restauração da atividade imunológica livre dos ataques à mielina (\"autoimune reset\"). Uma das maneiras de se monitorar a eficácia do tratamento é a identificação da ocorrência de novas lesões e de lesões com reforço visíveis em exames de RM seriados. Objetivo: Testar a hipótese de que o tratamento de pacientes com EM, utilizando AHSCT, foi eficaz em evitar a recorrência de inflamação e o aparecimento de novas lesões visíveis na SB ao exame de MRI, a longo prazo. Resultados: Na nossa Instituição, cerca de 66 pacientes portadores de EM foram tratados com aHSCT no período de 2004 a 2011, sendo seguidos desde então pelas disciplinas de hematologia, neurologia/neuroimunologia e pela seção de RM do CCIFM-HCRP. Método: Foram revisados os exames de RM de encéfalo adquiridos de maneira prospectiva e protocolada, de 76 pacientes submetidos ao aHSCT, com seguimento por MRI há mais de cinco anos. As imagens de RM foram arquivadas nos servidores do CCIFM, foram recuperadas, anonimizadas e revistas por pelo menos dois radiologistas experientes, de maneira independente e por confrontação. Foi considerada falha terapêutica a identificação de lesões novas e/ou lesões com reativação inflamatória. Resultados: Dez pacientes foram excluídos. Doze pacientes (18,18%) apresentaram novas lesões ou recorrência do processo inflamatório, com reforço. Conclusão: Na nossa amostra o aHSCT foi capaz de controlar a recorrência de lesões e da atividade inflamatória perceptível na RM em mais de 87% dos casos, caracterizando uma importante opção terapêutica de segunda linha nos casos de maior agressividade da doença / Multiple sclerosis (MS) is a recurrent chronic inflammatory demyelinating disease, restricted to the central nervous system (CNS), whose histological feature is the occurrence of perivenular inflammatory infiltrate, leading to demyelination with relative axonal preservation. The classic form of the disease is characterized by the recurrence of attacks (outbreaks), followed by remission of symptoms (RRMS), with the presence of multiple focal lesions dispersed by the CNS (spatial dissociation) with exuberant inflammatory process in the acute phase, coexisting with chronic lesions (Temporal dissociation) without inflammatory activity evidenced by the breakdown of blood-brain barrier and contrast-enhanced contrast-enhanced magnetic resonance imaging (MRI). Some patients have a benign course, remaining free of significant sequelae for more than 20 years of the disease. Other patients have an aggressive form of the disease, being restricted to the wheelchair in 8 to 10 years of evolution. One challenge is to modify the course in this aggressive way, which can be done with the use of immunomodulators, chemotherapeutics and, eventually, autologous hematopoietic stem cell transplantation (aHSCT). The goal of using aHSCT is to restore immune activity free of myelin attacks (\"autoimmune reset\"). One of the ways to monitor treatment efficacy is to identify the occurrence of new lesions and visible reinforcing lesions on serial MRI scans. Objective: To test the hypothesis that the treatment of patients with MS using AHSCT was effective in avoiding the recurrence of inflammation and the appearance of new visible lesions in SB at the long-term examination of MRI. Results: At our institution, approximately 66 patients with MS were treated with aHSCT from 2004 to 2011, followed by the hematology, neurology / neuroimmunology and MRI sections of the CCIFM-HCRP. Methods: Brain and MRI scans acquired in a prospective and protocolized way from 76 patients who underwent aHSCT were followed up with MRI for more than five years. The MR images were archived on the CCIFM servers, retrieved, anonymised and reviewed by at least two experienced radiologists, independently and by confrontation. The identification of new lesions and / or lesions with inflammatory reactivation was considered therapeutic failure. Results: Ten patients were excluded. Twelve patients (18.18%) presented new lesions or recurrence of the inflammatory process, with reinforcement. Conclusion: In our sample, aHSCT was able to control the recurrence of lesions and the inflammatory activity detected in MRI in more than 87% of the cases, characterizing an important second line therapeutic option in the cases of greater aggressiveness of the disease.

A esclerose múltipla (EM) é uma doença desmielinizante inflamatória crônica recorrente, restrita ao sistema nervoso central (SNC), cuja característica histológica é a ocorrência de desmielinização relacionada com infiltrado inflamatório perivenular com relativa preservação axonal. A forma clássica da doença se caracteriza pela recorrência de ataques (surtos), seguidos de remissão dos sintomas (RRMS), com a presença de múltiplas lesões focais dispersas pelo SNC (dissociação espacial) com processo inflamatório exuberante na fase aguda, coexistindo com lesões crônicas (dissociação temporal) sem atividade inflamatória evidenciavel pela quebra de barreira hematoencefálica e realce na fase contrastada na imagem de ressonância magnética (MRI). Alguns pacientes tem uma evolução benigna, permanecendo livre de sequelas significativas por mais de 20 anos da doença. Outros pacientes têm uma forma agressiva da doença, ficando restritos à cadeira de rodas em 8 a 10 anos de evolução. Um desafio é modificar o curso desta forma agressiva, o que pode ser feito com o uso de imunomoduladores, quimioterápicos e, eventualmente, transplante autólogo de células tronco hematopoiéticas (aHSCT). O objetivo da utilização do aHSCT é a restauração da atividade imunológica livre dos ataques à mielina (\"autoimune reset\"). Uma das maneiras de se monitorar a eficácia do tratamento é a identificação da ocorrência de novas lesões e de lesões com reforço visíveis em exames de RM seriados. Objetivo: Testar a hipótese de que o tratamento de pacientes com EM, utilizando AHSCT, foi eficaz em evitar a recorrência de inflamação e o aparecimento de novas lesões visíveis na SB ao exame de MRI, a longo prazo. Resultados: Na nossa Instituição, cerca de 66 pacientes portadores de EM foram tratados com aHSCT no período de 2004 a 2011, sendo seguidos desde então pelas disciplinas de hematologia, neurologia/neuroimunologia e pela seção de RM do CCIFM-HCRP. Método: Foram revisados os exames de RM de encéfalo adquiridos de maneira prospectiva e protocolada, de 76 pacientes submetidos ao aHSCT, com seguimento por MRI há mais de cinco anos. As imagens de RM foram arquivadas nos servidores do CCIFM, foram recuperadas, anonimizadas e revistas por pelo menos dois radiologistas experientes, de maneira independente e por confrontação. Foi considerada falha terapêutica a identificação de lesões novas e/ou lesões com reativação inflamatória. Resultados: Dez pacientes foram excluídos. Doze pacientes (18,18%) apresentaram novas lesões ou recorrência do processo inflamatório, com reforço. Conclusão: Na nossa amostra o aHSCT foi capaz de controlar a recorrência de lesões e da atividade inflamatória perceptível na RM em mais de 87% dos casos, caracterizando uma importante opção terapêutica de segunda linha nos casos de maior agressividade da doença / Multiple sclerosis (MS) is a recurrent chronic inflammatory demyelinating disease, restricted to the central nervous system (CNS), whose histological feature is the occurrence of perivenular inflammatory infiltrate, leading to demyelination with relative axonal preservation. The classic form of the disease is characterized by the recurrence of attacks (outbreaks), followed by remission of symptoms (RRMS), with the presence of multiple focal lesions dispersed by the CNS (spatial dissociation) with exuberant inflammatory process in the acute phase, coexisting with chronic lesions (Temporal dissociation) without inflammatory activity evidenced by the breakdown of blood-brain barrier and contrast-enhanced contrast-enhanced magnetic resonance imaging (MRI). Some patients have a benign course, remaining free of significant sequelae for more than 20 years of the disease. Other patients have an aggressive form of the disease, being restricted to the wheelchair in 8 to 10 years of evolution. One challenge is to modify the course in this aggressive way, which can be done with the use of immunomodulators, chemotherapeutics and, eventually, autologous hematopoietic stem cell transplantation (aHSCT). The goal of using aHSCT is to restore immune activity free of myelin attacks (\"autoimmune reset\"). One of the ways to monitor treatment efficacy is to identify the occurrence of new lesions and visible reinforcing lesions on serial MRI scans. Objective: To test the hypothesis that the treatment of patients with MS using AHSCT was effective in avoiding the recurrence of inflammation and the appearance of new visible lesions in SB at the long-term examination of MRI. Results: At our institution, approximately 66 patients with MS were treated with aHSCT from 2004 to 2011, followed by the hematology, neurology / neuroimmunology and MRI sections of the CCIFM-HCRP. Methods: Brain and MRI scans acquired in a prospective and protocolized way from 76 patients who underwent aHSCT were followed up with MRI for more than five years. The MR images were archived on the CCIFM servers, retrieved, anonymised and reviewed by at least two experienced radiologists, independently and by confrontation. The identification of new lesions and / or lesions with inflammatory reactivation was considered therapeutic failure. Results: Ten patients were excluded. Twelve patients (18.18%) presented new lesions or recurrence of the inflammatory process, with reinforcement. Conclusion: In our sample, aHSCT was able to control the recurrence of lesions and the inflammatory activity detected in MRI in more than 87% of the cases, characterizing an important second line therapeutic option in the cases of greater aggressiveness of the disease.