Integrative study of non-coding RNAs and metabolome profile in obese patients: pathophysiology mechanism contribution to search druggable targets

Abstract

Obesity prevalence has increased dramatically in at least threefold in last three decades becoming a very important problem in health public. Obesity is epidemiologically determinate as a high-risk factor for developing diseases, including cardiovascular diseases (CVDs). The complexity multifactor etiology of these diseases needs the application of Integrative tool interplaying environmental factors, lifestyle, and genetic/epigenetic factors in the pathophysiology mechanism to have a novel view of perspectives to determine early diagnosis, classification of new biomarkers, and discovery of potential targets for therapeutic interventions. Molecular mechanisms of obesity and cardiometabolic complications, still not well known, emerging the need for new study approach. Integrative analysis of epigenetic markers, as miRNAs and lcnRNAs that interfere in transcription profile and complemented with metabolomic profile may help to evaluate physiopathology of several complex diseases and can find out some useful biomarker and therapeutic target. The aim of this study is to propose an integrative analysis of some biological system including transcriptional regulation mechanism using expression of miRNA, lncRNA profile, mRNA and metabolomic profile in visceral adipose tissue (VAT) of obese individuals and lean controls. Also determine cardiometabolic profile (lipids, glucose metabolism, some inflammatory cytokines) and anthropometric profile with total fat body measurement. Moreover, miRNA and obesity-related lncRNA expression will be evaluated in peripheral leukocytes, and metabolomic and cardiometabolic profile will be evaluated in serum from a large number of obese patients and controls aiming to propose new useful biomarkers to predict risk of cardiovascular complications in obesity. VAT will be obtained from obese patients (BMI>30 Kg/m2) and lean controls (BMI 18-25 Kg/m2) of the Obesity Treatment Center at the Clinica Alemana de Temuco (CTO/CAT, Temuco-Chile) during an elective and clinically indicated laparoscopic surgery (bariatric surgery for patients and gallstones for controls). The RNA extracted from VAT will be used to evaluate the miRNA profile by next generation sequencing, mRNA profile of cholesterol metabolism by RT-qPCR, lncRNA related to cholesterol metabolism and metabolomic profile using mass spectrometry acoplated to UHPLC. Results will be evaluated by integrative bioinformatic and machine learning computing modelling tools to select the main molecules related to obesity and cardiometabolic complications. These molecules will be tested as biomarkers in peripheral leukocytes from a larger cohort from obese subjects (n=125) enrolled in Real Benemérita Associação Portuguesa Hospital (Sao Paulo, Brazil) and at CTO/CAT-Chile, using RT-qPCR. Serum metabolomic analysis will be also performed. Identification of functional features will be performed bioinformatic tools will be employed to identify potential druggable targets using molecular modeling. The results of epigenomic and metabolomic mechanisms will contribute for knowledge of molecular bases in the pathophysiology of obesity and its cardiovascular complications. Moreover, in clinical practice, it has potential application as biomarkers that can be validated as diagnosis and follow-up of therapy and search for therapeutic alternatives. (AU)