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STUDY OF PROTEIN BIOMARKERS AND GENE EXPRESSION PROFILE OF NF-˜B PATHWAY IN LATE-ONSET NEONATAL SEPSIS: POTENTIAL VALUE IN DIAGNOSIS AND PROGNOSIS

Grant number: 16/06887-5
Support type:Regular Research Grants
Duration: February 01, 2017 - January 31, 2020
Field of knowledge:Health Sciences - Medicine - Maternal and Child Health
Principal Investigator:Patricia Palmeira Daenekas Jorge
Grantee:Patricia Palmeira Daenekas Jorge
Home Institution: Faculdade de Medicina (FM). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Assoc. researchers:Andréia Cristiane Rangel Santos ; Fernanda Andrade Macaferri da Fonseca ; Magda Maria Sales Carneiro-Sampaio ; Maria Esther Jurfest Rivero Ceccon ; Maria Helena Baptista Nunes da Silva ; Werther Brunow de Carvalho

Abstract

The availability of a panel of protein or genetic biomarkers that can differentiate sepsis from Systemic Inflammatory Response Syndrome (SIRS) earlier and also identify different response patterns caused by different etiological agents, may cause significant impact on morbidity and mortality of neonatal sepsis, contributing to the reduction of hospital costs and also differentiate a good or bad prognosis. The purpose is to describe the protein and genetic expression of selected biomarkers in term newborns (NBs) with clinical late-onset sepsis and / or culture-proved caused by Gram-positive bacteria, Gram-negative bacteria and fungi at the day of diagnosis (day 0), in the intermediate stage (3rd day) and in the convalescence phase (7th day). Blood samples will be collected from 108 NBs distributed in groups according to the etiological agent: NBs with sepsis caused by Gram-positive bacteria (Group 1, n = 27); caused by Gram-negative bacteria (group 2, n = 27); caused by fungi (Group 3, n = 27) and; NBs with clinical late-onset sepsis (group 4, n = 27). Blood samples from 27 neonates distributed among the four study groups will be collected in the healing phase to compose the control group. Soluble protein biomarkers sTREM, IL-27, MBL, CX3CL1, Hepcidin, Endocan and IL-18 will be evaluated by ELISA, as well as CD64 expression on the surface of neutrophils and monocytes by flow cytometry. For gene expression, next generation sequencing will be performed to assess RNA expression of genes related to nuclear factor kappa B (NF-8B) signaling pathway using the customized Kit TruSeq Targeted RNA Expression NF-™B panel to include the genes from TREM-1, IL-27, IL-18, MBL2, CX3CL1, HAMP (hepcidina) e ESM1 (Endocan) (Illumina Inc.). Statistical analysis will be performed by GraphPad Prism 5.0 software and in addition to the descriptive statistics, parametric and non-parametric tests will be performed in accordance with the distribution of data. p-values < 0.05 will be considered statistically significant. (AU)