MicroRNAs Signature Panel Identifies Heavy Drinkers with Alcohol-Associated Cirrhosis from Heavy Drinkers without Liver Injury

Fathima Shihana; Mugdha V Joglekar; Tae-Hwi Schwantes-An; Anandwardhan A Hardikar; Devanshi Seth

doi:10.3390/biology12101314

MicroRNAs Signature Panel Identifies Heavy Drinkers with Alcohol-Associated Cirrhosis from Heavy Drinkers without Liver Injury

Biology (Basel). 2023 Oct 8;12(10):1314. doi: 10.3390/biology12101314.

Authors

Fathima Shihana^{1

2}, Mugdha V Joglekar³, Tae-Hwi Schwantes-An⁴, Anandwardhan A Hardikar³, Devanshi Seth^{1

2

5}

Affiliations

¹ The Centenary Institute of Cancer Medicine & Cell Biology, The University of Sydney, Sydney, NSW 2006, Australia.
² Edith Collins Centre (Translational Research in Alcohol Drugs and Toxicology), Sydney Local Health District, Sydney, NSW 2050, Australia.
³ Diabetes & Islet Biology Group, School of Medicine, Western Sydney University, Campbelltown, NSW 2560, Australia.
⁴ Department of Medical and Molecular Genetics, Indiana University, Indianapolis, IN 46202, USA.
⁵ Sydney School of Medicine, Faculty of Medicine and Health, The University of Sydney, Sydney, NSW 2006, Australia.

Abstract

Background: Alcohol-associated liver disease (ALD) is the most common disorder of prolonged drinking. Mechanisms underlying cirrhosis in such patients remain unclear. MicroRNAs play regulatory role in several diseases, are affected by alcohol and may be important players in alcohol use disorders, such as cirrhosis. Methods: We investigated serum samples from heavy chronic alcohol users (80 g/day (male) and 50 g/day (female) for ≥10 years) that were available from our previously reported GenomALC study. A subset of GenomALC drinkers with liver cirrhosis (cases, n = 24) and those without significant liver disease (drinking controls, n = 23) were included. Global microRNA profiling was performed using high-throughput real-time quantitative PCR to identify the microRNA signatures associated with cirrhosis. Ingenuity Pathway Analysis (IPA) software was utilized to identify target mRNAs of significantly altered microRNAs, and molecular pathways were analysed. Identified microRNAs were analysed for correlation with traditional liver disease biomarkers and risk gene variants previously reported from GenomALC genome-wide association study. Results: The expression of 21 microRNAs was significantly downregulated in cases compared to drinking controls (p < 0.05, ∆∆Ct > 1.5-fold). Seven microRNAs (miR-16, miR-19a, miR-27a, miR-29b, miR-101, miR-130a, and miR-191) had a highly significant correlation (p < 0.001) with INR, bilirubin and MELD score. Three microRNAs (miR-27a, miR-130a and miR-191) significantly predicted cases with AUC-ROC 0.8, 0.78 and 0.85, respectively (p < 0.020); however, INR performed best (0.97, p < 0.001). A different set of six microRNAs (miR-19a, miR-26a, miR-101, miR-151-3p, miR-221, and miR-301) showed positive correlation (ranging from 0.32 to 0.51, p < 0.05) with rs10433937:HSD17B13 gene variant, associated with the risk of cirrhosis. IPA analysis revealed mRNA targets of the significantly altered microRNAs associated with cell death/necrosis, fibrosis and increased steatosis, particularly triglyceride metabolism. Conclusions: MicroRNA signatures in drinkers distinguished those with liver cirrhosis from drinkers without liver disease. We identified mRNA targets in liver functions that were enriched for disease pathogenesis pathways.

Keywords: alcohol-associated liver cirrhosis; biomarker; microRNAs.

Grants and funding

U01 AA018389/AA/NIAAA NIH HHS/United States