Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance
Katherine Johnson, Peter Leary, Olivier Govaere, Matthew J. Barter, Sarah Charlton, Simon Cockell, Dina Tiniakos, Michalina Zatorska, Pierre Bédossa, M. Julia Brosnan, Jeremy Cobbold, Mattias Ekstedt, Guruprasad P. Aithal, Karine Clément, Jörn M. Schattenberg, Jérôme Boursier, Vlad Ratziu, Elisabetta Bugianesi, Quentin M. Anstee, Ann K. Daly, James Clark, Heather J. Cordell, Rebecca Darlay, Christopher P. Day, Timothy Hardy, Yang‐Lin Liu, Fiona Oakley, Jeremy M. Palmer, Rachel Queen, Kristy Wonders, Patrick M. Bossuyt, Adriaan G. Holleboom, Hadi Zafarmand, Yasaman Vali, Jenny Lee, Karine Clément, Raluca Pais, Detlef Schuppan, Michael Allison, Sergio Rodriguez Cuenca, Vanessa Pellegrinelli, Michèle Vacca, Antonio Vidal‐Puig, Tuulia Hyötyläinen, Aidan McGlinchey, Matej Orešič, Partho Sen, José M. Mato, Óscar Millet, Jean‐François Dufour, Stephen A. Harrison, Stefan Neubauer, Michael Pavlides, Ferenc E. Mózes, Salma Akhtar, Rajarshi Banerjee, Matt Kelly, Elizabeth Shumbayawonda, Andrea Dennis, Charlotte Erpicum, Manuel Romero‐Gómez, Rocío Gallego‐Durán, Isabel Fernández-Lizaranzu, M.A. Karsdal, Diana Leeming, Mette Juul Fisker, Elisabeth Erhardtsen, Daniel Guldager Kring Rasmussen, Per Qvist, Antonia Sinisi, Estelle Sandt, Maria Manuela Tonini, Maurizio Parola, Chiara Rosso, Fabio Marra, Amalia Gastaldelli, Sven Francque, Stergios Kechagias, Hannele Yki‐Järvinen, Kimmo Porthan, Saskia W. C. van Mil, George Papatheodoridis, Helena Cortez‐Pinto, Luca Valenti, Salvatore Petta, Luca Miele, Andreas Geier, Christian Trautwein, Paul Hockings, Philip N. Newsome, David Wenn, Cecília M. P. Rodrigues, Rémy Hanf, Pierre Chaumat, Christian Rosenquist, Aldo Trylesinski, Pablo A. Ortiz, Kevin L. Duffin, Carla Yunis, Melissa Miller
Abstract
Background & Aims: Serum microRNA (miRNA) levels are known to change in non-alcoholic fatty liver disease (NAFLD) and may serve as useful biomarkers. This study aimed to profile miRNAs comprehensively at all NAFLD stages. Methods: We profiled 2,083 serum miRNAs in a discovery cohort (183 cases with NAFLD representing the complete NAFLD spectrum and 10 population controls). miRNA libraries generated by HTG EdgeSeq were sequenced by Illumina NextSeq. Selected serum miRNAs were profiled in 372 additional cases with NAFLD and 15 population controls by quantitative reverse transcriptase PCR. Results: Levels of 275 miRNAs differed between cases and population controls. Fewer differences were seen within individual NAFLD stages, but miR-193a-5p consistently showed increased levels in all comparisons. Relative to NAFL/non-alcoholic steatohepatitis (NASH) with mild fibrosis (stage 0/1), 3 miRNAs (miR-193a-5p, miR-378d, and miR378d) were increased in cases with NASH and clinically significant fibrosis (stages 2-4), 7 (miR193a-5p, miR-378d, miR-378e, miR-320b, miR-320c, miR-320d, and miR-320e) increased in cases with NAFLD activity score (NAS) 5-8 compared with lower NAS, and 3 (miR-193a-5p, miR-378d, and miR-378e) increased but 1 (miR-19b-3p) decreased in steatosis, activity, and fibrosis (SAF) activity score 2-4 compared with lower SAF activity. The significant findings for miR-193a-5p were replicated in the additional cohort with NAFLD. Studies in Hep G2 cells showed that following palmitic acid treatment, miR-193a-5p expression decreased significantly. Gene targets for miR-193a-5p were investigated in liver RNAseq data for a case subgroup (n = 80); liver GPX8 levels correlated positively with serum miR-193a-5p. Conclusions: Serum miR-193a-5p levels correlate strongly with NAFLD activity grade and fibrosis stage. MiR-193a-5p may have a role in the hepatic response to oxidative stress and is a potential clinically tractable circulating biomarker for progressive NAFLD. Lay summary: MicroRNAs (miRNAs) are small pieces of nucleic acid that may turn expression of genes on or off. These molecules can be detected in the blood circulation, and their levels in blood may change in liver disease including non-alcoholic fatty liver disease (NAFLD). To see if we could detect specific miRNA associated with advanced stages of NAFLD, we carried out miRNA sequencing in a group of 183 patients with NAFLD of varying severity together with 10 population controls. We found that a number of miRNAs showed changes, mainly increases, in serum levels but that 1 particular miRNA miR-193a-5p consistently increased. We confirmed this increase in a second group of cases with NAFLD. Measuring this miRNA in a blood sample may be a useful way to determine whether a patient has advanced NAFLD without an invasive liver biopsy.