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Serum Coding and Non-Coding RNAs as Biomarkers of NAFLD and Fibrosis Severity

43 Pages Posted: 28 Sep 2018

See all articles by Stefania Di Mauro

Stefania Di Mauro

University of Catania - Department of Clinical and Experimental Medicine

Alessandra Scamporrino

University of Catania - Department of Clinical and Experimental Medicine

Salvatore Petta

University of Palermo - Section of Gastroenterology and Hepatology

Francesca Urbano

University of Catania - Department of Clinical and Experimental Medicine

Agnese Filippello

University of Catania - Department of Clinical and Experimental Medicine

Marco Ragusa

University of Catania - Genome and Complex Systems BioMedicine; Associazione Oasi Maria S.S. - Institute for Research on Mental Retardation and Brain Aging (I.R.C.C.S.)

Maria Teresa Di Martino

Magna Graecia University of Catanzaro - Department of Experimental and Clinical Medicine

Francesca Scionti

Magna Graecia University of Catanzaro - Department of Experimental and Clinical Medicine

Stefania Grimaudo

University of Palermo - Section of Gastroenterology and Hepatology

Rosaria Maria Pipitone

University of Palermo - Biomedical Department of Internal and Specialized Medicine (DI.BI.M.I.S.)

Graziella Privitera

University of Catania - Department of Clinical and Experimental Medicine

Antonino Di Pino

University of Catania - Department of Clinical and Experimental Medicine

Roberto Scicali

University of Catania - Department of Clinical and Experimental Medicine

Agata Maria Rabuazzo

University of Catania - Department of Clinical and Experimental Medicine

Antonio Craxì

University of Palermo - Section of Gastroenterology and Hepatology

Michele Purrello

University of Catania - Genome and Complex Systems BioMedicine

Francesco Purrello

University of Catania - Department of Clinical and Experimental Medicine

Salvatore Piro

University of Catania - Department of Clinical and Experimental Medicine

More...

Abstract

Background: In patients with NAFLD liver biopsy is the gold standard to detect NASH and stage liver fibrosis. We aimed to identify differentially expressed mRNAs and non-coding RNAs in serum samples of biopsy-diagnosed mild and severe NAFLD patients with respect to controls and to each other.  

Methods: We first performed a whole transcriptome analysis through microarray (n=12: 4 CTRL; 4 mild NAFLD: NAS≤4 F0; 4 severe NAFLD NAS≥5 F3), followed by validation of selected transcripts through qPCRs in 88 subjects: 63 NAFLD, 25 CTRL. A similar analysis was also performed on HepG2 exposed to Oleate:Palmitate or only Palmitate (cellular model of NAFL/NASH) at intracellular/extracellular levels. Transcript correlation with histological/clinical data was also analyzed.  

Findings: We identified several DE coding/non-coding RNAs in each group of the study cohort. We validated the up-regulation of UBE2V1, BNIP3L mRNAs, and TGFB2/TGFB2-OT1 coding/ lncRNA both in patients with NAS ≥5 (versus NAS ≤ 4) and in patients with Fibrosis stages= 3-4 (versus F= 0-2). HBA2 mRNA and RP11-128N14.5 lncRNA were up-regulated respectively only in F=3-4 or NAS ≥5 patients. Analyzed transcript deregulation was also reflected in NAFL/NASH in vitro models. UBE2V1, RP11-128N14.5, BNIP3L, and TGFB2/TGFB2-OT1 correlated with histological/biochemical data. Combinations of TGFB2/TGFB2-OT1 FIB-4 (AUC=0.891, p-value=0.000003) or TGFB2/TGFB2-OT1 Fibroscan (AUC=0.892, p-value=0.000002) improved the detection of F=3-4 with respect to F=0-2 fibrosis stages.  

Interpretation: We identified specific serum coding/non-coding RNA profiles in severe and mild NAFLD patients that possibly mirror molecular mechanisms underlying NAFLD progression towards NASH/fibrosis. TGFB2/TGFB2-OT1 detection improves FIB-4/Fibroscan diagnostic performance for advanced fibrosis discrimination.

Funding: This study was funded by 2016/2018 Department Research Plan of University of Catania, Department of Clinical and Experimental Medicine (project #A) and by 2016/2018 Department Research Plan of University of Catania (2nd line of intervention).

Declaration of Interest: The Authors do not have any conflict of interest to declare.

Ethical Approval:  This study was conducted according to the Declaration of Helsinki and was approved by the ethics committee of the University Hospital of Palermo. Before the procedures a written informed consent was obtained from all patients participating in this study.

Keywords: NASH, FIBROSIS, RNA, HepG2, DIAGNOSIS, LIQUID-BIOPSY

Suggested Citation

Di Mauro, Stefania and Scamporrino, Alessandra and Petta, Salvatore and Urbano, Francesca and Filippello, Agnese and Ragusa, Marco and Di Martino, Maria Teresa and Scionti, Francesca and Grimaudo, Stefania and Pipitone, Rosaria Maria and Privitera, Graziella and Di Pino, Antonino and Scicali, Roberto and Rabuazzo, Agata Maria and Craxì, Antonio and Purrello, Michele and Purrello, Francesco and Piro, Salvatore, Serum Coding and Non-Coding RNAs as Biomarkers of NAFLD and Fibrosis Severity (September 13, 2018). Available at SSRN: https://ssrn.com/abstract=3249697 or http://dx.doi.org/10.2139/ssrn.3249697

Stefania Di Mauro

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Alessandra Scamporrino

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Salvatore Petta

University of Palermo - Section of Gastroenterology and Hepatology ( email )

Palermo
Italy

Francesca Urbano

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Agnese Filippello

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Marco Ragusa

University of Catania - Genome and Complex Systems BioMedicine

Catania
Italy

Associazione Oasi Maria S.S. - Institute for Research on Mental Retardation and Brain Aging (I.R.C.C.S.)

United States

Maria Teresa Di Martino

Magna Graecia University of Catanzaro - Department of Experimental and Clinical Medicine

Catanzaro, 88100
Italy

Francesca Scionti

Magna Graecia University of Catanzaro - Department of Experimental and Clinical Medicine

Catanzaro, 88100
Italy

Stefania Grimaudo

University of Palermo - Section of Gastroenterology and Hepatology

Palermo
Italy

Rosaria Maria Pipitone

University of Palermo - Biomedical Department of Internal and Specialized Medicine (DI.BI.M.I.S.)

Palermo
Italy

Graziella Privitera

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Antonino Di Pino

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Roberto Scicali

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Agata Maria Rabuazzo

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy

Antonio Craxì

University of Palermo - Section of Gastroenterology and Hepatology ( email )

Palermo
Italy

Michele Purrello

University of Catania - Genome and Complex Systems BioMedicine

Catania
Italy

Francesco Purrello (Contact Author)

University of Catania - Department of Clinical and Experimental Medicine ( email )

Via Palermo, 636
Catania, Catania 95122
Italy

Salvatore Piro

University of Catania - Department of Clinical and Experimental Medicine

Via Palermo, 636
Catania, Catania 95122
Italy