TY - JOUR
T1 - Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression
T2 - Diagnostic and mechanistic relevance
AU - Johnson, Katherine
AU - Leary, Peter J.
AU - Govaere, Olivier
AU - Barter, Matthew J.
AU - Charlton, Sarah H.
AU - Cockell, Simon J.
AU - Tiniakos, Dina
AU - Zatorska, Michalina
AU - Bedossa, Pierre
AU - Brosnan, M. Julia
AU - Cobbold, Jeremy F.
AU - Ekstedt, Mattias
AU - Aithal, Guruprasad P.
AU - Clément, Karine
AU - Schattenberg, Jörn M.
AU - Boursier, Jerome
AU - Ratziu, Vlad
AU - Bugianesi, Elisabetta
AU - Anstee, Quentin M.
AU - Daly, Ann K.
AU - Clark, James
AU - Cordell, Heather J.
AU - Darlay, Rebecca
AU - Day, Christopher P.
AU - Hardy, Tim
AU - Liu, Yang Lin
AU - Oakley, Fiona
AU - Palmer, Jeremy
AU - Queen, Rachel
AU - Wonders, Kristy
AU - Bossuyt, Patrick M.
AU - Holleboom, Adriaan G.
AU - Zafarmand, Hadi
AU - Vali, Yasaman
AU - Lee, Jenny
AU - Clement, Karine
AU - Pais, Raluca
AU - Schuppan, Detlef
AU - Allison, Michael
AU - Cuenca, Sergio Rodriguez
AU - Pellegrinelli, Vanessa
AU - Vacca, Michele
AU - Vidal-Puig, Antonio
AU - Hyötyläinen, Tuulia
AU - McGlinchey, Aidan
AU - Orešič, Matej
AU - Sen, Partho
AU - Mato, Jose
AU - Millet, Óscar
AU - Sandt, Estelle
AU - LITMUS Consortium Investigators
N1 - Funding Information:
This study has been performed as part of the LITMUS (Liver Investigation: Testing Marker Utility in Steatohepatitis) project, which has received funding from the Innovative Medicines Initiative (IMI2) Program of the European Union under Grant Agreement 777377; this Joint Undertaking receives support from the European Union's Horizon 2020 research and innovation programme and EFPIA. The current study was also supported by the Newcastle NIHR Biomedical Research Centre and the European NAFLD Registry.
Publisher Copyright:
© 2021 The Author(s)
PY - 2022/2
Y1 - 2022/2
N2 - 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.
AB - 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.
KW - Biomarker
KW - MicroRNA
KW - Non-alcoholic fatty liver disease
KW - Sequencing
UR - http://www.scopus.com/inward/record.url?scp=85122616328&partnerID=8YFLogxK
UR - https://pubmed.ncbi.nlm.nih.gov/35072021
U2 - 10.1016/j.jhepr.2021.100409
DO - 10.1016/j.jhepr.2021.100409
M3 - Article
C2 - https://pubmed.ncbi.nlm.nih.gov/35072021
AN - SCOPUS:85122616328
SN - 2589-5559
VL - 4
SP - 100409
JO - JHEP Reports
JF - JHEP Reports
IS - 2
M1 - 100409
ER -