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Authors
Abstract(s)
Sarcomas são um grupo heterogéneo de tumores malignos com origem na transformação de células mesenquimais nos tecidos conjuntivos. Estes representam menos de 1% dos cancros adultos e abrangem mais de 170 subtipos, dos quais aproximadamente 87% provêm de tecidos moles e 13% do osso.
A carcinogénese depende de alterações metabólicas e apresenta características comuns a um vasto espetro de diferentes cancros, tal como evasão à apoptose, potencial replicativo ilimitado, reprogramação do metabolismo energético, entre outros.
Dois substratos metabólicos comummente utilizados pelas células cancerígenas para satisfazer as exigências biossintéticas e bioenergéticas são a glucose e a glutamina. Uma forma de estudar as características metabólicas dos tumores, incluindo os sarcomas, abrange diversas técnicas que serão descritas nesta Tese. A resultante Metabolómica é uma ampla análise quantitativa e qualitativa do metaboloma e uma das vantagens desta abordagem reside no facto dos metabolitos estarem a jusante de genes, transcrição e proteínas e, portanto, oferecem a representação mais próxima do fenótipo.
Neste projeto, tínhamos como objetivo criar uma coleção de amostras de sarcoma no Biobanco do iMM-CAML, bem como caracterizar metabolicamente estas amostras, utilizando espectroscopia de RMN e avaliação da expressão genética por PCR quantitativo.
Os resultados preliminares obtidos por RMN apresentaram diferenças evidentes entre o tecido tumoral e o tecido saudável adjacente. As amostras tumorais indicaram uma alta atividade glicolítica, apresentando níveis elevados de lactato acompanhados por níveis reduzidos de glucose. Além disso, foram demonstrados níveis elevados de glutamato juntamente com níveis mais baixos de glutamina, o que sugere um metabolismo de glutamina mais elevado nestas amostras. Outras diferenças observadas envolvem níveis de glucose-1-fosfato e UDP-glucose, bem como BCAA, aminoácidos aromáticos e carnosina. No que diz respeito à expressão genética, os resultados obtidos mostraram uma expressão mais elevada de LDHA, seguida por GAPDH e ALDOC. A GLS2, envolvida no metabolismo da glutamina, mostra o nível mais baixo de expressão entre os genes avaliados. A continuação da caracterização metabolómica de amostras de STS permitirá o estabelecimento de diferenças discriminativas entre STS e tecido normal, contribuindo para o desenvolvimento de terapias mais eficazes para estes tumores.
Sarcomas are a heterogenous group of malignant tumors that originate from the transformation of mesenchymal cells in connective tissues. These account for less than 1% of adult cancers and encompass over 170 subtypes, of which approximately 87% arise from soft tissue and 13% from bone. Carcinogenesis is dependent on alterations of the metabolic activity and presents certain characteristics common to a wide spectrum of different cancers, such as apoptosis evasion, unlimited replicative potential, reprogramming of energy metabolism, among others. Two metabolic substrates commonly used by cancer cells to meet the biosynthetic and bioenergetic requirements of these cells are glucose and glutamine. One way to study the metabolic characteristics of tumors, including sarcomas, comprises several techniques that will be described in this thesis. The resulting Metabolomics is a broad quantitative and qualitative analysis of the metabolome and one of the main advantages regarding this approach lies on the fact that metabolites are downstream of genes, transcripts, and proteins and therefore, give the closest representation of the phenotype. In this project, we aimed to create a collection of sarcoma samples at the biobank of iMM-CAML, as well as metabolically characterize these samples using NMR spectroscopy and evaluation of genetic expression by quantitative PCR. The preliminary results obtained on 1H NMR profiling of STS displayed evident differences between tumoral tissue and adjacent non-involved healthy tissue. Tumor samples indicated high glycolytic activity by displaying higher levels of lactate alongside lower levels of glucose. Furthermore, increased levels of glutamate were shown together with lower levels of glutamine, which suggests a higher glutamine metabolism in these samples. Other prominent differences observed regard levels of glucose-1-phosphate and UDP-glucose as well as BCAA, aromatic amino acids and carnosine. Regarding gene expression, results obtained showed a higher expression of LDHA, followed by GAPDH and ALDOC. GLS2, involved in glutamine metabolism, shows the lowest level of expression amongst the assessed genes. The extension of metabolomic characterization of STS samples will allow the establishment of discriminative differences between STS and normal tissue, contributing to the development of more effective therapies for these tumors.
Sarcomas are a heterogenous group of malignant tumors that originate from the transformation of mesenchymal cells in connective tissues. These account for less than 1% of adult cancers and encompass over 170 subtypes, of which approximately 87% arise from soft tissue and 13% from bone. Carcinogenesis is dependent on alterations of the metabolic activity and presents certain characteristics common to a wide spectrum of different cancers, such as apoptosis evasion, unlimited replicative potential, reprogramming of energy metabolism, among others. Two metabolic substrates commonly used by cancer cells to meet the biosynthetic and bioenergetic requirements of these cells are glucose and glutamine. One way to study the metabolic characteristics of tumors, including sarcomas, comprises several techniques that will be described in this thesis. The resulting Metabolomics is a broad quantitative and qualitative analysis of the metabolome and one of the main advantages regarding this approach lies on the fact that metabolites are downstream of genes, transcripts, and proteins and therefore, give the closest representation of the phenotype. In this project, we aimed to create a collection of sarcoma samples at the biobank of iMM-CAML, as well as metabolically characterize these samples using NMR spectroscopy and evaluation of genetic expression by quantitative PCR. The preliminary results obtained on 1H NMR profiling of STS displayed evident differences between tumoral tissue and adjacent non-involved healthy tissue. Tumor samples indicated high glycolytic activity by displaying higher levels of lactate alongside lower levels of glucose. Furthermore, increased levels of glutamate were shown together with lower levels of glutamine, which suggests a higher glutamine metabolism in these samples. Other prominent differences observed regard levels of glucose-1-phosphate and UDP-glucose as well as BCAA, aromatic amino acids and carnosine. Regarding gene expression, results obtained showed a higher expression of LDHA, followed by GAPDH and ALDOC. GLS2, involved in glutamine metabolism, shows the lowest level of expression amongst the assessed genes. The extension of metabolomic characterization of STS samples will allow the establishment of discriminative differences between STS and normal tissue, contributing to the development of more effective therapies for these tumors.
Description
Tese de mestrado, Ciências Biofarmacêuticas, 2022, Universidade de Lisboa, Faculdade de Farmácia.
Keywords
Cancer Sarcoma Metabolomic Metabolic remodelling Teses de mestrado - 2022