Banca de DEFESA: TAYLA DA CRUZ SANTOS PEREIRA

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
DISCENTE : TAYLA DA CRUZ SANTOS PEREIRA
DATA : 29/07/2020
HORA: 09:00
LOCAL: https://conferenciaweb.rnp.br/webconf/defesa-de-dissertacao-tayla-pereira-pmbqbm
TÍTULO:

METABOLOMIC ANALYSIS OF THE PLASMA OF INDIVIDUALS WITH OSTEONECROSIS SECONDARY TO SICKLE CELL DISEASE

PALAVRAS-CHAVES:

Sickle cell disease, osteonecrosis, metabolomics, nuclear magnetic resonance.

PÁGINAS: 152
GRANDE ÁREA: Ciências Biológicas
ÁREA: Bioquímica
SUBÁREA: Metabolismo e Bioenergética
RESUMO:

Sickle cell disease is the most common hemoglobinopathy in Brazil and constitutes a worldwide public health problem, with a great impact on the morbidity and mortality of the affected population. In patients with this pathology, the predominant joint clinical manifestation is osteonecrosis, which commonly progresses to terminal disease. Analyzes related to metabolites and unregulated metabolic pathways in complex systems have become a powerful tool for investigating metabolic processes, identifying potential biomarkers and unraveling metabolic reprogramming in various diseases. In this context, this work had as main objective to use a metabolic approach by nuclear magnetic resonance to identify biomarkers related to sickle cell disease, as well as to osteonecrosis secondary to sickle cell disease. Blood plasma from control subjects and patients with sickle cell disease with and without osteonecrosis was used. Based on the clinical stage of osteonecrosis, graduated using the modified Ficat and Arlet method, the samples were classified as early, intermediate and advanced stages of osteonecrosis. To obtain the uni and bidimensional spectra, a nuclear magnetic resonance equipment - Bruker Avance III of 14.1 Tesla - (600 MHz) was used. The optimization of the sample preparation and spectrum acquisition protocol involved the use of different pulse sequences; Zg, Zgpr, Lcipnf2, Noesypr 1D as well as, some protocols for protein precipitation with methanol. The identification of the metabolites was performed with the aid of the Chenomx NMR Suite 8.4 software. Processing and quantification were performed using the NMRProcflow software. Statistical analyzes were performed using MetaboAnalyst 3.0 software. Twenty-nine metabolites were identified by 1H NMR, including 12 amino acids, 9 organic acids, 4 lipids, 3 organic compounds, 1 carbohydrate. The metabolic profile of the blood plasma of the control group was significantly different from the sickle cell groups without osteonecrosis and the sickle cell group with osteonecrosis. Twenty metabolites with VIP score > 0.5 were responsible for the differences between samples in these groups. Sn-glycerol-3-phosphocholine and phenylalanine were the metabolites with the most promising results for future investigations of the sickle cell disease marker. 3-hydroxybutyrate and VLDL + HDL were the metabolites with the most promising results for future studies of osteonecrosis biomarker secondary to DF. The discriminant analysis also revealed that the plasma of patients with osteonecrosis secondary to DF at different stages of osteonecrosis has different metabolic profiles. In the peripheral blood plasma, fourteen metabolites with VIP score > 0.5 were responsible for this difference. Histidine and lactate were the metabolites with the most promising results for future investigations of osteonecrosis staging biomarkers secondary to sickle cell disease, especially for the early diagnosis of this pathology in peripheral blood plasma. Histidine, VLDL + HDL and citrate were the metabolites with the most promising results for future investigations of biomarker of advanced stages of osteonecrosis secondary to sickle cell disease in peripheral blood plasma. Fifteen metabolites with VIP score > 0.5 were classified as the set of metabolites
responsible for the differences between the bone marrow plasma of individuals in different stages of osteonecrosis secondary to sickle cell disease. Citrate was the metabolite with the most promising result for future investigations of osteonecrosis staging biomarkers secondary to sickle cell disease, especially for the early diagnosis of this pathology in bone marrow plasma. Lactate was the metabolite with the most promising result for future investigations of biomarker of advanced stages of osteonecrosis secondary to sickle cell disease in bone marrow plasma. The topological analysis of metabolic pathways revealed a potential relationship between sickle cell disease and osteonecrosis secondary to DF and some metabolic pathways, namely; metabolism of nitrogen, pyruvate, thiamine, arginine and proline, phenylalanine; biosynthesis of aminoacyl t-RNA, phenylalanine, tyrosine and tryptophan, valine, leucine and isoleucine; glycolysis or gluconeogenesis; degradation of valine, leucine and isoleucine; synthesis and degradation of ketone bodies. These results provide strong evidence of a metabolic signature for individuals with sickle cell disease and osteonecrosis secondary to DF, defined mainly by a set of twenty metabolites with altered blood plasma levels including, proline, lactate, creatine + creatinine, 2-hydroxy -3-methylbutyric, phenylalanine, glycine, acetone, format, citrate, glucose, trimethylamione-n-oxide, histidine, tyrosine, azelate, 3-hydroxybutyrate, leucine + isoleucine, sn-glycerol-3-phosphocoline and acetate. It is hoped that these findings will help to guide future research in the area and may further elucidate the biochemical changes in sickle cell disease and osteonecrosis secondary to DF.

MEMBROS DA BANCA:
Presidente - 3768242 - PAULO ROBERTO RIBEIRO DE JESUS
Externo ao Programa - 2341233 - ELISANGELA VITORIA ADORNO
Externo à Instituição - ANTONIO GILBERTO FERREIRA - UFSCAR