Aplicação da metabolômica multiplataforma (HPLC-MS, CE-MS E GC-MS) non-targeted em estudos biomédicos
Data
2019-12-18
Autores
Furtado, Danielle Zildeana Sousa 
Orientadores
Assunção, Nilson Antonio de
Tipo
Tese de doutorado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
A aplicação da Metabolômica em estudos biomédicos é um campo da área da saúde que vem ganhando credibilidade, devido a obtenção de informações bioquímicas relevantes sobre o estado fisiológico ou patológico do organismo. A associação deste campo com técnicas multiplataformas aumentam a capacidade de leitura da assinatura bioquímica das doenças, devido a maior cobertura do metaboloma. A aplicação da metabolômica associada a técnicas de alto desempenho já é uma realidade em alguns laboratórios, das quais auxiliam no diagnóstico de doenças. Entretanto, nem todas as doenças têm a sua assinatura bioquímica esclarecida, como exemplo as doenças raras: Síndrome de Cri du Chat, principalmente retratando a cardiopatia e a doença de Behçet, que são consideradas raras devido ao grande número de sinais e sintomas. A síndrome de Cri du Chat é uma anomalia cromossômica, resultante da deleção distal ou intertiscial do braço curto do cromossomo 5. As principais características desta condição genética são: o choro de recém-nascidos semelhante ao miado agudo de um gato, alterações neurológicas, motoras, cardíacas etc. A síndrome de Behçet é uma vasculite mutisistêmica; cuja principais características clínicas são: lesões orais, genitais, inflamação ocular etc. Seguindo este fluxo de informações, foi proposto neste estudo a análise Metabolômica Non-targeted utilizando ferramentas multiplataforma para a análise global, a fim de verificar alterações bioquímicas de cada enfermidades. As amostras de plasma foram coletadas e analisada por HPLC-MS, CE-MS e GC-MS, que juntamente com a aplicação de métodos quimiometricos (PCA, PLS-DA) e da estatística univariada (Teste T) foram possíveis a identificação de alterações de metabólitos compatíveis com o metabolismo dos glicerofosfolipídeos em ambas enfermidades raras. Além disso foram encontrados os metabólitos relacionados com o desenvolvimento da aterosclerose e de problemas cardíacos coronários (Síndrome de Cri du chat) e metabólitos relacionados ao sistema imune (Síndrome de Behçet). Estes resultados são pré-eliminares e acredita-se que a associação da metabolômica com outros estudos ômicos poderá ajudar no esclarecimento do fisio-bioquimico destas síndromes.
Metabolomic’s application in biomedical studies is a field of health that has been gaining credibility due to obtaining relevant biochemical information about the physiological or pathological state of the organism. The association of this field with multiplatform techniques increases the reading capacity of the biochemical signature of diseases, due to the greater coverage of the metabolome. The application of metabolomics associated with high performance techniques is already a reality in some laboratories, which help in the diagnosis of diseases. However, not all diseases have their biochemical signature clarified, such as rare diseases: Cri du Chat syndrome, especially heart disease and Behçet's, which are considered rare due to the large number of signs and symptoms. Cri du Chat syndrome is a chromosomal anomaly resulting from distal or interstitial deletion of the short arm of chromosome 5. The main features of this genetic condition are: newborn crying similar to a cat's acute meow, neurological, motor changes, cardiac, etc. Behçet's syndrome is a multisystemic vasculitis; whose main clinical features are: oral, genital lesions, eye inflammation, etc. Following this information flow, it was proposed in this study the Non-targeted Metabolomic analysis using multiplatform tools for the non-targeted analysis, in order to verify biochemical alterations of each disease. Plasma samples were collected and analyzed by HPLC-MS, CE-MS and GC-MS, which together with the application of chemometric tools (PCA, PLS-DA) and univariate statistics (T-Test) were possible to identify compatible changes with glycerophospholipid metabolism in both rare syndromes. In addition, metabolites related to the development of atherosclerosis and coronary heart problems (Cri du chat syndrome) and metabolites related to the immune system (Behçet's syndrome) were found. These results are pre-eliminated, and it is believed that the association of metabolomics with other omic studies may help clarify the physio-biochemical of these diseases.
Metabolomic’s application in biomedical studies is a field of health that has been gaining credibility due to obtaining relevant biochemical information about the physiological or pathological state of the organism. The association of this field with multiplatform techniques increases the reading capacity of the biochemical signature of diseases, due to the greater coverage of the metabolome. The application of metabolomics associated with high performance techniques is already a reality in some laboratories, which help in the diagnosis of diseases. However, not all diseases have their biochemical signature clarified, such as rare diseases: Cri du Chat syndrome, especially heart disease and Behçet's, which are considered rare due to the large number of signs and symptoms. Cri du Chat syndrome is a chromosomal anomaly resulting from distal or interstitial deletion of the short arm of chromosome 5. The main features of this genetic condition are: newborn crying similar to a cat's acute meow, neurological, motor changes, cardiac, etc. Behçet's syndrome is a multisystemic vasculitis; whose main clinical features are: oral, genital lesions, eye inflammation, etc. Following this information flow, it was proposed in this study the Non-targeted Metabolomic analysis using multiplatform tools for the non-targeted analysis, in order to verify biochemical alterations of each disease. Plasma samples were collected and analyzed by HPLC-MS, CE-MS and GC-MS, which together with the application of chemometric tools (PCA, PLS-DA) and univariate statistics (T-Test) were possible to identify compatible changes with glycerophospholipid metabolism in both rare syndromes. In addition, metabolites related to the development of atherosclerosis and coronary heart problems (Cri du chat syndrome) and metabolites related to the immune system (Behçet's syndrome) were found. These results are pre-eliminated, and it is believed that the association of metabolomics with other omic studies may help clarify the physio-biochemical of these diseases.
Descrição
Citação
FURTADO, Danielle Zildeana Sousa. Aplicação da metabolômica multiplataforma (HPLC-MS, CE-MS E GC-MS) non-targeted em estudos biomédicos. 2019. 158 f. Tese (Doutorado em Medicina Translacional) - Escola Paulista de Medicina, Universidade Federal de São Paulo (UNIFESP). São Paulo, 2019.