Integration of proteomic and metabolomic analyses: New insights for mapping informal workers exposed to potentially toxic elements

Occupational exposure to potentially toxic elements (PTEs) is a concerning reality of informal workers engaged in the jewelry production chain that can lead to adverse health effects. In this study, untargeted proteomic and metabolomic analyses were employed to assess the impact of these exposures on informal workers' exposome in Limeira city, São Paulo state, Brazil. PTE levels (Cr, Mn, Ni, Cu, Zn, As, Cd, Sn, Sb, Hg, and Pb) were determined in blood, proteomic analyses were performed for saliva samples (n = 26), and metabolomic analyses in plasma (n = 145) using ultra-high performance liquid chromatography (UHPLC) coupled with quadrupole-time-of-flight (Q-TOF) mass spectrometry. Blood PTE levels of workers, controls, and their family members were determined by inductively coupled plasma-mass spectrometry (ICP-MS). High concentration levels of Sn and Cu were detected in welders' blood (p < 0.001). Statistical analyses were performed using MetaboAnalyst 4.0. The results showed that 26 proteins were upregulated, and 14 proteins downregulated on the welder group, and thirty of these proteins were also correlated with blood Pb, Cu, Sb, and Sn blood levels in the welder group (p < 0.05). Using gene ontology analysis of these 40 proteins revealed the biological processes related to the upregulated proteins were translational initiation, SRP-dependent co-translational protein targeting to membrane, and viral transcription. A Metabolome-Wide Association Study (MWAS) was performed to search for associations between blood metabolites and exposure groups. A pathway enrichment analysis of significant features from the MWAS was then conducted with Mummichog. A total of 73 metabolomic compounds and 40 proteins up or down-regulated in welders were used to perform a multi-omics analysis, disclosing seven metabolic pathways potentially disturbed by the informal work: valine leucine and isoleucine biosynthesis, valine leucine and isoleucine degradation, arginine and proline metabolism, ABC transporters, central carbon metabolism in cancer, arachidonic acid metabolism and cysteine and methionine metabolism. The majority of the proteins found to be statistically up or downregulated in welders also correlated with at least one blood PTE level, providing insights into the biological responses to PTE exposures in the informal work exposure scenario. These findings shed new light on the effects of occupational activity on workers' exposome, underscoring the harmful effects of PTE

Dysregulation of glycerophospholipid metabolism during Behçet's disease contributes to a pro-inflammatory phenotype of circulating monocytes

Behcet's disease (BD) is a relapsing, multisystem and inflammatory condition characterized by systemic vasculitis of small and large vessels. Although the etiopathogenesis of BD remains unknown, immune-mediated mechanisms play a major role in the development of the disease. BD patients present leukocyte infiltration in the mucocutaneous lesions as well as neutrophil hyperactivation. In contrast to neutrophils, whose involvement in the pathogenesis of BD has been extensively studied, the biology of monocytes during BD is less well known. In this study, we analyzed the phenotype and function of circulating monocytes of 38 BD patients from Hospital of Braga. In addition, we evaluated the impact of inflammatory and metabolomic plasma environment on monocyte biology. We observed a worsening of mitochondrial function, with lower mitochondrial mass and increased ROS production, on circulating monocytes of BD patients. Incubation of monocytes from healthy donors with the plasma of BD patients mimicked the observed phenotype, strongly suggesting the involvement of serum mediators. BD patients, regardless of their symptoms, had higher serum pro-inflammatory TNF-alpha and IP-10 levels and IL-1 beta/IL-1RA ratio. Untargeted metabolomic analysis identified a dysregulation of glycerophospholipid metabolism on BD patients, where a significant reduction of phospholipids was observed concomitantly with an increase of lysophospholipids and fatty acids. These observations converged to an enhanced phospholipase A2 (PLA(2)) activation. Indeed, inhibition of PLA(2) with dexamethasone or the downstream cyclooxygenase (COX) enzyme with ibuprofen was able to significantly revert the mitochondrial dysfunction observed on monocytes of BD patients. Our results show that the plasma inflammatory environment coupled with a dysregulation of glycerophospholipid metabolism in BD patients contribute to a dysfunction of circulating monocytes

Estudos bioanalíticos aplicados à busca do perfil de metabólitos em portadores da síndrome cri du chat

Introdução: A síndrome de Cri Du Chat é uma doença rara, caracterizada por choro semelhante ao miado de gato, resultante de uma deleção na região do braço curto do cromossomo 5. Objetivo: Efetuar a caracterização do perfil dos metabólitos em pacientes portadores da Síndrome Cri-Du-Chat comparando a indivíduos com metabolismo normal de uma mesma família, através da análise metabolômica e abordagem Quimiométrica. Metodologia: As amostras do plasma sanguíneo e da urina foram coletadas de pessoas de ambos os sexos com idade (1 a 38 anos), coletado no total de 38 amostras de plasma e urina, posteriormente, analisadas por ultra cromatografia líquida acoplada ao espectrômetro de massas, eletroforese capilar, cromatografia de íons, conseguinte foi aplicado o teste estatístico de Wilcoxon e uma abordagem quimiométrica para as interpretações dos resultados do ponto de vista bioquímico. Resultados: Houve um aumento plasmático dos níveis da metionina-sufóxido, aspartato e serotonina; e diminuição dos níveis da citosina, histidina, isoleucina, leucina, ornitina, fenilalanina, valina, creatinina, também foi avaliado o aumento da atividade do superóxido dismutase e diminuição dos tíos císteina e glutationa. Na urina foram observados o aumento das concentrações dos seguintes metabólitos: alanina, asparagina, aspartato, citrulina, glutamina, histidina, fenilalanina, serina, treonina, tirosina, triptofano, metionina sufóxido e sarcosina. Conclusões: Os portadores da desordem genética apresentaram alterações metabolômicas, que podem estar relacionados em processos de estresse oxidativo, alterações pirimídicas, ciclo da uréia, ciclo do ácido cítrico e alterações na biossíntese de aminoácidos, via serotonérgica e modificações no processo do catabolimos e anabolismo.Dados abertos - Sucupira - Teses e dissertações (2013 a 2016

Aplicação da metabolômica multiplataforma (HPLC-MS, CE-MS E GC-MS) non-targeted em estudos biomédicos

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.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.Dados abertos - Sucupira - Teses e dissertações (2019