IN SILICO IDENTIFICATION AND IN VIVO VALIDATION OF NON-INVASIVE BIOMARKERS FOR LIVER FIBROSIS

GIRAUDI PABLO; GAMBARO SABRINA ELIANA; CHACKELEVICIUS CARLA; GIURICIN M.; CROCE LORY; BONAZZA D; SOARDO G.; DE MANZINI N.; TIRIBELLI CLAUDIO; PALMISANO S; ROSSO NATALIA

Bacelona

Congreso; The International Liver Congress? 2016, EASL; 2016

European Association for the Study of the Liver, EASL

Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is aprogressive    liver    injury,    which    can    lead    to   non-alcoholicsteatohepatitis   and   progress   towards   cirrhosis.   It   is   tightlyassociated with obesity and type 2 diabetes mellitus. Currently, thegold standard for the diagnosis relies on the liver biopsy. Reliablenon-invasive tools are still missing. The aim of this study was toidentify  novel  biomarkers  by  in  silico  studies  and  validate  theirspecificity in serum of a NAFLD cohort.Methods: in silico protein-protein interactions were used to build abiological network among interested genes involved in fibrogenesis(CK18, IBP3, LCP1, LGALS1, LAMB1, HACTA, TIMP-2 and MMP2). Fromthis  analysis  were  identified:  CD44,  SPARC,  EGFR  and  IGF2,  assecreted  factors  with  clinical  relevance  as  candidate  biomarkers.Gene expression of interest and candidate genes were assessed bothin liver and blood by qPCR. Plasmatic concentration of candidates wasmeasured by ELISA.45  adult  morbid  obese  consenting  patients  (BMI > 35 kg/m2;  17males  and  28  females,  age  range:  19?63)  undergoing  bariatricsurgery  were  enrolled.  Wedge  liver  biopsy  was  performed  andblood  samples  were  collected.  Exclusion  criteria  were  alcoholconsumption, viral hepatitis infection, known chronic liver disease.Biopsy was assessed according to Brunt?s fibrosis Score.Results: Hepatic gene expression analysis showed an increase of thegenes  of  interest,  which  correlated  with  the  stage  of  fibrosis.Interestingly, the expression of candidates genes: IGF2, CD44 andSPARC,  also  increased  with  the  fibrosis?  stage.  They  allowed  todistinguish grade 0 vs. 2 of liver fibrosis (folds of expression: 1.0 ± 0.3vs.12.1 ± 4.0; 1.0 ± 0.5 vs. 4.9 ± 2.2 and 1.0 ± 0.4 vs 11.4 ± 4.4, p < 0.05, p< 0.05  and  p < 0.01,  respectively).  Only  CK18  hepatic  expressioncorrelated   with   both   steatosis   grade   and   fibrosis   stage.   Nocorrelations between liver and blood gene expression was observedfor any studied gene. IGF2 plasmatic level was inversely correlatedwith fibrosis score (p < 0.01), whereas EGFR levels increased withfibrosis score (p < 0.01).Conclusions: These preliminary data show that plasma levels of theidentified soluble markers (IGF2 and EGFR) seem to correlate withthe  initial  stage  of  fibrosis.  This  data  could  contribute  to  thedevelopment of future non-invasive diagnosis tools.U05SPFRA14 ? FRA 2014 (CdA dd. 19.12.2014) FIF grant. PJG wassponsored by Fondazione Umberto Veronesi (Grants 2015). SEG byProyectonutrizione 297 CTGAS CMC by MAE.