Cytokine storm in the covid-19 patient: contribution of adipose tissue

Hypothesis: The white adipose tissue (WAT) contributes, as one of the main endocrine glands, to the cytokine storm and the systemic effects of the infection.Justification: i) WAT is an important component of the inflammatory response in cachexia, a systemic disease in which hypoxia, inflammation and fibrosis occur in the main organs. Therefore, it is possible to expect similarities, given the recent description of the effects of Covid19 on organs / tissues19 ii) ii) WAT secretes all major inflammatory cytokines, eicosanoids and also adipokines. There is no literature available on the expression of inflammatory / autacoid cytokines and adipokines in patients with WAT from COVID19 Objectives: 1. Examine the morphology of WAT, immune infiltration, fibrosis and expression of inflammatory factors 2. Investigate intracellular NFKB (adipocyte and monocyte) and inflammatory pathways 3. Study the load of exosomes derived from WAT. Casuistry and experiments: a) SAMPLES: WAT (subcutaneous and visceral deposits), plasma University Hospital (HU-USP): 40 patients with positive Covid-19 test, submitted to emergency surgery; 20 negative patients for the Covid19 tests, who underwent the same surgeries Faculdade de Medicina-USP: samples obtained from autopsies of patients who passed Covid19 (20); equal number of control samples b) Experiments: WAT: ¾ Histological analysis of tissue (light and electron microscopy) ¾ FACS exam study of infiltrated immune cells ¾ Multiplex assays for inflammatory cytokines, proteins involved in hypoxia, proteins related to fibrosis pathways PCR PCR in real time for the same factors and microRNA analysis ¾ Western blot for evaluation of phosphorylated proteins in the pathways of interest Lip Lipidomics of Incub tissues Incubation of explants and separation of exosomes and subsequent analysis (lipidomics, miRs ) Plasma: ¾ Inflammatory / metabolic profile: quantification of circulating markers of inflammation, cardiometabolic disease, neuropeptides, adipokines and hormones (insulin, cortisol, testosterone, estradiol) PCR, cytokines, neuropeptides, adipokines, glucose, lactate, creatinine, insulin, cortisol Plasma / lipidomic metabolomic metabolism; Separation Exosome separation and load evaluation (AU) quantification of circulating markers of inflammation, cardiometabolic disease, neuropeptides, adipokines and hormones (insulin, cortisol, testosterone, estradiol) PCR, cytokines, neuropeptides, adipokines, glucose, lactate, creatinine, insulin, cortisol Metabol of plasma / lipid metabolism; Separation Exosome separation and load assessment (AU) quantification of circulating markers of inflammation, cardiometabolic disease, neuropeptides, adipokines and hormones (insulin, cortisol, testosterone, estradiol) PCR, cytokines, neuropeptides, adipokines, glucose, lactate, creatinine, insulin, cortisol Metabol of plasma / lipid metabolism; Separation Exosome separation and load assessment (AU)