Estudo metabolômico para descoberta de biomarcadores no soro de pacientes com doença de Parkinson

Abstract

Parkinson's disease (PD) is a neurodegenerative disease with a complex etiology that mainly affects the elderly. The diagnosis is still based on the clinical symptoms and treatment options are limited. In this context, it is urgent to search for biomarkers that can help in the diagnosis in a faster and more accurate way. In addition, it is important to determine affected metabolic pathways for research into new therapeutic options. Therefore, the objective of this work was to evaluate, through metabolomic strategies, the variations in the level of metabolites in the serum of patients with PD to identify biomarkers for diagnosis and the regulated metabolic pathways. Thus, the blood of the participants (patient and control group) was collected, centrifuged and the serum withdrawn. After preparing the samples, they were analyzed by ultra-performance liquid chromatography (UPLC) coupled to a quadrupole-time-of-flight mass spectrometer with electrospray ionization (ESI) (UPLC-ESI-QTOF-MS). The results obtained were processed in the MasslynxsTM software and later analyzed in the SIMCAP + 12.0 software for supervised multivariate statistical analysis. The most significant variables for projection (VIPs) (>2) in this analysis were unreplicated through the HMDB (Human Metabolome Database) platform. The data were also processed in the MS-DIAL software and analyzed in the Metaboanalyst online tool using univariate and multivariate statistical analyses, considering VIPs >1 for dereplication using the MS-FINDER. Subsequently, an analysis of altered metabolic pathways was performed. Through statistical analysis, a welldefined separation between the control and PD patient groups was observed, and that the analytical method was satisfactorily reproducible. It was possible to dereplicate metabolites correlated with the patient and correlated with the control group and being referenced with a DP, such as 15(S)-HPETE, 12-KETE, pipecolic acid and 1-lyso-2-arachidonoylphosphatidate. Also, it was possible to identify alterations in the metabolism pathways of sphingolipids, glutathione, arachidonic acid, caffeine metabolism, arachidonic acid and primary bile acid biosynthesis. Therefore, the results allowed us to point out important information about possible PD biomarkers and the metabolic mechanisms involved in the pathology as a support for the discovery of markers for the diagnosis and therapeutic targets.